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Pea protein is a promising ingredient for plant-based cheese production but has poor consumer acceptance due to intrinsic beany flavors. Fermentation could potentially decrease these off-flavors while also producing desirable cheese-like aromas. Pea protein emulsion gels were fermented using four different bacterial blends for 16 weeks with and without the crosslinking enzyme transglutaminase. The volatile organic compound (VOC) profiles were assessed by GC-MS and the peptide profile was measured by LC-MS/MS during storage. VOC production was mainly affected by the composition of the bacterial blends, followed by storage time. Crosslinking of the protein gel structure had minimal impact on VOC production. The peptide-level profiling revealed that crosslinking can reduce peptide size and the production of bitterness-like peptides in some blends. This study provides insights into the effect of bacterial blends, storage time, and enzymatic crosslinking on the production of volatile components and peptides related to aroma and peptide profiles for pea protein., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Biodegradation of polyurethane (PU) plastics is a lower cost and more environmentally friendly approach to the regeneration of waste plastics than the landfill or incineration alternatives. Currently, however, the lack of efficient degradation strains and their enzymes is restricting the development of viable large-scale waste PU regeneration. In this study, a wild strain (LTX1) is isolated from a coastal mudflat, and then a mutant strain (MLTX1) with higher degradation efficiency is obtained by UV mutagenesis. Both the LTX1 and MLTX1 strains are able to achieve a more than 80 % weight loss of PU foam after 12 days treatment, making them the most efficient PU foam-degrading strains available to date. The PU foam degradation is characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). A novel gene, purh, encoding one of the cutinases is cloned using genomics and transcriptomics, and its recombinant PurH, capable of efficiently degrading PU foam, is expressed in Escherichia coli and identified. The discovery of this highly-efficient PU foam-degrading strain and its enzyme may represent a leap forward in the biological depolymerization and recycling of PU foam., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Stone fruits and their processed products are highly valued in the whole world for their flavor, aroma, rich nutritional contents, and various health benefits. While large quantities of stone fruits are produced globally, significant losses occur due to improper handling and storage, from production to consumption. This review focuses on the application of advanced non-thermal treatment techniques for whole fresh stone fruits and their processed products. It provides a comprehensive assessment of the factors contributing to spoilage, along with the mechanisms, applications, and limitations of non-thermal techniques in reducing spoilage. Compared to traditional preservation methods, such as the use of artificial food additives, chemicals, thermal treatments, and low-temperature storage, these novel techniques demonstrate better results in minimizing spoilage. Moreover, non-thermal techniques are most sustainable and eco-friendly, as they reduce energy consumption, minimize chemical use, and generate less waste than traditional methods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A multifunctional oxidase-mimicking Ag/Mn 3 O 4 was prepared, catalyzing the 3, 3', 5, 5'-tetramethylbenzidine (TMB) chromogenic reaction. Six foodborne pathogenic bacteria species, including Escherichia coli, Staphylococcus aureus, Salmonella enterica, Listeria monocytogenes, Bacillus cereus, and Cronobacter sakazakii, were observed to differentially inhibit its oxidase-like activity, resulting in decelerating the TMB chromogenic reaction. Owing to these properties, the following achievements were achieved: colorimetric determination of these bacteria with high sensitivity can be achieved using Ag/Mn 3 O 4  + TMB reaction system; precise identification of these bacteria at different concentrations, including individual bacterium, binary mixtures, and even multivariate mixtures, can be effectively realized by combining the Ag/Mn 3 O 4 -based colorimetric sensor array with principal component analysis (PCA); broad-spectrum inactivation of these bacteria can be remarkably realized through catalyzation of Ag/Mn 3 O 4 to generate superoxide anion free radicals. Therefore, our proposed Ag/Mn 3 O 4 holds significant application potential in the colorimetric determination, precise identification, and broad-spectrum inactivation of foodborne pathogenic bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In recent years, a number of synthetic potentiators of antibiotics have been discovered. Their action can significantly enhance the antibacterial effect and limit the spread of antibiotic resistance through inhibition of bacterial cystathionine-γ-lyase. To expand the known set of potentiators, we developed methods for the synthesis of five new representatives of 6-bromoindole derivatives-potential inhibitors of bacterial cystathionine-γ-lyase-namely potassium 3-amino-5-((6-bromoindolyl)methyl)thiophene-2-carboxylate ( MNS2 ) and its 6-bromoindazole analogs ( MNS3 and MNS4 ), along with two 6-broindazole analogs of the parent compound NL2 . Their syntheses are based on 6-bromoindole, 6-bromoindazole and methyl 5-(bromomethyl)-3-((ethoxycarbonyl)amino)thiophene-2-carboxylate as the main building blocks, assembling the rest of the heterocyclic system on their basis at the nitrogen atom. We assessed the ability of the new inhibitors to potentiate the antimicrobial activity of gentamicin.

Glycosaminoglycans (GAGs), as animal polysaccharides, are linked to proteins to form various types of proteoglycans. Bacterial GAG lyases are not only essential enzymes that spoilage bacteria use for the degradation of GAGs, but also valuable tools for investigating the biological function and potential therapeutic applications of GAGs. The ongoing discovery and characterization of novel GAG lyases has identified an increasing number of lyases suitable for functional studies and other applications involving GAGs, which include oligosaccharide sequencing, detection and removal of specific glycan chains, clinical drug development and the design of novel biomaterials and sensors, some of which have not yet been comprehensively summarized. GAG lyases can be classified into hyaluronate lyases, chondroitinases and heparinases based on their substrate spectra, and their functional applications are mainly determined by their substrates, with different lyases exhibiting differing substrate selectivity and preferences. It is thus necessary to understand the properties of the available enzymes to determine strategies for their functional application. Building on previous studies and reviews, this Review highlights small yet crucial differences among or within the various GAG lyases to aid in optimizing their use in future studies. To clarify ideas and strategies for further research, we also discuss several traditional and novel applications of GAG lyases., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2025. Published by The Company of Biologists.)

This study has developed a pressure sensor array based on four functionalized DNA-nanoenzymes with catalase-like activity for multiple detections of foodborne pathogens through a portable pressure manometer. Benefiting from functionalization of 4-mercaptophenylboronic acid and β-mercaptoethylamine, the diversity of nonspecific interactions between four DNA-nanoenzymes and each of the nine bacteria leads to differences in pressure response patterns by catalyzing H 2 O 2 to generate exclusive "fingerprints". As effective statistical tools for processing multivariate data, principal component analysis and hierarchical clustering analysis are employed to identify nine foodborne pathogens by analyzing pressure response patterns. Furthermore, the as-prepared sensor array can discriminate different mixtures of bacteria and achieve quantitative detection, with an average detection limit of 10 2 and 10 4 CFU mL -1 for Gram-positive and Gram-negative bacteria, respectively, demonstrating its desirable practicality and satisfactory accuracy for real sample detection. This study expands insights into multiple analyses of foodborne pathogens for food safety monitoring.

Endogenous enzymes play a crucial role in determining fish product aroma. However, the attached microorganisms can promote enzyme production, making it challenging to identify specific aromatic compounds resulting from endogenous enzymes. Thus, we investigated the aroma transformation of Japanese sea bass through enzymatic incubation by controlling attached microorganisms during the lag phase. Our results demonstrate that enzymatic incubation significantly enhances grassy and sweet notes while reducing fishy odors. These changes in aroma are associated with increased levels of 10 volatile compounds and decreased levels of 3 volatile compounds. Among them, previous studies have reported enzyme reaction pathways for octanal, 1-nonanal, vanillin, indole, linalool, geraniol, citral, and 6-methyl-5-hepten-2-one; however, the enzymatic reaction pathways for germacrene D, beta-caryophyllene, pristane, 1-tetradecene and trans-beta-ocimene remain unclear. These findings provide novel insights for further study to elucidate the impact of endogenous enzymes on fish product aromas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The focus on microalgae for applications in several fields, e.g. resources for biofuel, the food industry, cosmetics, nutraceuticals, biotechnology, and healthcare, has gained increasing attention over the last decades. In this study, we investigate the microbiome of the cultured microalga Tetraselmis chui (T. chui) to highlight their potential for health benefits. In this context, biomolecules like antioxidants play a crucial role in the well-being of living organisms as they metabolise harmful reactive oxygen species (ROS) to reduce oxidative stress. Impaired processing of ROS leads to damaged cells and increases the risk of cancer, inflammatory diseases, and diabetes, among others. Here, we identify, characterise, and test bacterial antioxidants derived from the T. chui microbiome metagenome dataset. We identified 258 genes coding for proteins with potential antioxidant activity. Of those, four novel enzymes are expressed and identified as two superoxide dismutases (SOD), TcJM_SOD2 and TcIK_SOD3, and two catalases (CAT), TcJM_CAT2 and TcIK_CAT3. Extensive analyses characterised all implemented enzymes as active even in concentrations down to 25 ng*ml -1 for the SODs and 15 ng*ml -1 for the CATs. Furthermore, sequence-based analyses assign TcJM_SOD2 and TcIK_SOD3 to iron superoxide dismutases (Fe SODs) and TcJM_CAT2 and TcIK_CAT3 to heme-containing catalases. These candidates are phylogenetically classified within the phylum Pseudomonadota. Regarding the biotechnological potential, a toxicity assay did not indicate any harmful effects. The introduced enzymes may benefit medical applications and expand the potential of microalgae microbiomes. KEY POINTS: • Omics-based discoveries of antioxidant enzymes from Tetraselmis chui microbiome • Two superoxide dismutases and two catalases are identified and tested for activity • Enzyme sensitivity highlights biotechnological potential of microalgae microbiomes., Competing Interests: Declarations. Ethical approval: The manuscript presents research on Galleria mellonella that do not require ethical approval for their study. Conflict of interest: The authors declare no competing interests., (© 2025. The Author(s).)

Enzyme-based depolymerization of plastics, including polyesters, has emerged as a promising approach for plastic waste recycling and reducing environmental plastic pollution. Currently, most of the known polyester-degrading enzymes are represented by a few natural and engineered PETases from the carboxylesterase family V. To identify novel groups of polyesterases, we selected 25 proteins from the carboxylesterase family IV, which share 22 % to 80 % sequence identity to the metagenomic thermophilic polyesterase IS12. All purified proteins were found to be active against chromogenic para-nitrophenyl esters with a preference for short acyl chains. Screening for polyesterase activity using emulsified polyesters demonstrated the presence of hydrolytic activity against bis(benzoyloxyethyl) terephthalate (3PET), polycaprolactone (PCL), and polylactic acid (PLA) in all tested proteins. Biochemical characterization of four selected polyesterases revealed high thermostability in CBA10055, whereas the mesophilic GEN0105 exhibited higher polyesterase activity. Two ancestral variants of GEN0105 showed higher thermostability and activity against PCL and PLA, but reduced activity with amorphous PET. Furthermore, six established PETases were found to be highly active against PCL and PLA. Thus, our results indicate that polyesterase activity is widespread in the family IV carboxylesterases, and that most polyesterases are promiscuous being able to degrade different polyesters., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Thermophilic microbial communities growing in low-oxygen environments often contain early-evolved archaea and bacteria, which hold clues regarding mechanisms of cellular respiration relevant to early life. Here, we conducted replicate metagenomic, metatranscriptomic, microscopic, and geochemical analyses on two hyperthermophilic (82-84 °C) filamentous microbial communities (Conch and Octopus Springs, Yellowstone National Park, WY) to understand the role of oxygen, sulfur, and arsenic in energy conservation and community composition. We report that hyperthermophiles within the Aquificota (Thermocrinis), Pyropristinus (Caldipriscus), and Thermoproteota (Pyrobaculum) are abundant in both communities; however, higher oxygen results in a greater diversity of aerobic heterotrophs. Metatranscriptomics revealed major shifts in respiratory pathways of keystone chemolithotrophs due to differences in oxygen versus sulfide. Specifically, early-evolved hyperthermophiles express high levels of high-affinity cytochrome bd and CydAA' oxidases in suboxic sulfidic environments and low-affinity heme Cu oxidases under microaerobic conditions. These energy-conservation mechanisms using cytochrome oxidases in high-temperature, low-oxygen habitats likely played a crucial role in the early evolution of microbial life., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Predicting how tooth and dental material bonds perform in the mouth requires a deep understanding of degrading factors. Yet, this understanding is incomplete, leading to significant uncertainties in designing and evaluating new dental adhesives. The durability of dental bonding interfaces in the oral microenvironment is compromised by bacterial acids, salivary enzymes, and masticatory fatigue. These factors degrade the bond between dental resins and tooth surfaces, making the strength of these bonds difficult to predict. Traditionally studied separately, a combined kinetic analysis of these interactions could enhance our understanding and improvement of dental adhesive durability. To address this issue, we developed and validated an original model to evaluate the bond strength of dental restorations using realistic environments that consider the different mechanical, chemical, and biological degradative challenges working simultaneously: bacteria, salivary esterases, and cyclic loading. We herein describe a comprehensive investigation on dissociating the factors that degrade the bond strength of dental restorations. Our results showed that cariogenic bacteria are the number one factor contributing to the degradation of the bonded interface, followed by cyclic loading and salivary esterases. When tested in combinatorial mode, negative and positive synergies towards the degradation of the interface were observed. Masticatory loads (i.e., cycling loading) enhanced the lactic acid bacterial production and the area occupied by the biofilm at the bonding interface, resulting in more damage at the interface and a reduction of 73 % in bond strength compared to no-degraded samples. Salivary enzymes also produced bond degradation caused by changes in the chemical composition of the resin/adhesive. However, the degradation rates are slowed compared to the bacteria and cyclic loading. These results demonstrate that our synergetic model could guide the design of new dental adhesives for biological applications without laborious trial-and-error experimentation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Macrocyclization of peptides reduces conformational flexibilities, potentially leading to improved drug-like properties. However, side reactions such as epimerization and oligomerization often pose synthetic challenges. Peptide-cyclizing biocatalysts in the biosynthesis of non-ribosomal peptides (NRPs) have remarkable potentials as chemoenzymatic tools to facilitate more straightforward access to complex macrocycles. This review highlights the biocatalytic potentials of NRP cyclases, especially those of cis-acting thioesterases, the most general cyclizing machinery in NRP biosynthesis. Growing insights into penicillin-binding protein-type thioesterases, a relatively new group of trans-acting thioesterases, are also summarized., Competing Interests: Declarations. Conflict of interest: There are no conflicts to declare., (© 2024. The Author(s).)

Pyrite-driven autotrophic denitrification (PAD) has been recognized as a promising treatment technology for nitrate removal. Although the occurrence of PAD has been found in recent years, there is a knowledge gap about effects of crystal plane of pyrite on the performance and mechanism of PAD system. Here, this study investigated the effects of crystal planes ({100}, {111} and {210}) of single-crystal pyrite on denitrification performance, electron transfer, and microbial mechanism in PAD system. The removal efficiency of nitrate in B-{210} reached 100%, which was 1.67-fold and 2.86-fold higher than that of B-{100} and B-{111}, respectively. X-ray photoelectron spectroscopy and electrochemical results indicated that Fe-S bonds of pyrite with {210} crystal plane were more susceptible to breakage by Fe 3+ oxidation assault, and leaching microbially available Fe 2+ and sulfur intermediates to drive autotrophic denitrification. Metagenomic results suggested that community of functional pyrite-driven denitrifiers varied in response to crystal plane, and abundances of N-S transformation and EET-related microbes and genes in B-{210} notably up-regulated compared to B-{100} and B-{111}. In addition, this work proposed a dual-mode for electron transfer pathway during pyrite oxidation and nitrogen transformation in PAD system. In B-{210}, Fe(II)- and sulfur-driven denitrifiers obtained electron after pyrite oxidation-dissolution, and the enrichment of pyrite-oxidizing bacteria in B-{210} could enhance the electron transfer from pyrite through electron shuttles. This work highlighted that stronger surface reactivity and electron shuttle effect in B-{210} enhanced electron transfer, leading to favorable PAD performance in B-{210}. Overall, this study provides novel insights into the structure-activity relationship between the crystal plane structure of pyrite and denitrification activity in PAD system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

While microbial phosphate removal in activated sludge (AS) systems has been extensively studied, the role of viruses in this process remains largely unexplored. In this study, we identified 149 viral auxiliary metabolic genes associated with phosphorus cycling from 2,510 viral contigs (VCs) derived from AS systems. Notably, polyphosphate kinase 1 (ppk1) and polyphosphate kinase 2 (ppk2) genes, which are primarily responsible for phosphate removal, were found in five unclassified VCs. These genes exhibited conserved protein structures and active catalytic sites, indicating a pivotal role of viruses in enhancing phosphorus removal. Phylogenetic analysis demonstrated a close relationship between viral ppk genes and their bacterial counterparts, suggesting the occurrence of horizontal gene transfer. Furthermore, experimental assays validated that viral ppk genes enhanced host phosphate removal capabilities. VCs carrying ppk genes were observed across diverse ecological and geographical contexts, suggesting their potential to bolster host functions in varied environmental and nutrient settings, spanning natural and engineered systems. These findings uncover a previously underappreciated mechanism by which viruses enhance phosphate removal in wastewater treatment plants. Overall, our study highlights the potential for leveraging virus-encoded genes to improve the efficiency of biological phosphorus removal processes, offering new insights into the microbial ecology of AS systems and the role of viruses in biogeochemical cycling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Fatty acids are used in fundamental cellular processes, such as membrane biogenesis, energy generation, post-translational modification of proteins, and so forth. These processes require the activation of fatty acids by adenosine triphosphate (ATP), followed by condensation with coenzyme-A (CoA), catalyzed by the omnipresent enzyme called Fatty acyl-CoA ligases (FACLs). However, Fatty acyl-AMP ligases (FAALs), the structural homologs of FACLs, operate in an unprecedented CoA-independent manner. FAALs transfer fatty acids to the acyl carrier protein (ACP) domain of polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS) for the biosynthesis of various antibiotics, lipopeptides, virulent complex lipids, and so forth in bacteria. Recent structural and biochemical insights from our group provide a detailed understanding of the mode of CoA rejection and ACP acceptance by FAALs. In this review, we have discussed advances in the mechanistic, evolutionary, and functional understanding of FAALs and FAAL-like domains across life forms. Here, we are proposing a "Five-tier" mechanistic model to explain the specificity of FAALs. We further demonstrate how FAAL-like domains have been repurposed into a new family of proteins in eukaryotes with a novel function in lipid metabolism., (© 2023 Wiley Periodicals LLC.)

Studies of enzymes catalyzing carbon-fluorine (C-F) bond cleavage have focused largely on a limited number of native microbial hydrolases that are reactive with the natural product fluoroacetate. Driven by widespread interest in biodegrading commercial fluorinated compounds, many of which are known as per- and polyfluorinated alkyl substances (PFAS), it is necessary to identify and engineer new enzymes. For example, some hydrolases react with -CF 2 - moieties, a common functionality in PFAS. Additional enzymatic C-F cleaving mechanisms catalyzed by reductases, lyases, and oxygenases have been identified via screening. Screening and evolving PFAS defluorination in bacteria is inhibited by the obligate release of toxic fluoride from C-F cleavage. Engineering greater fluoride tolerance in bacteria is a problem that must be solved in tandem with enzyme improvement., Competing Interests: Declaration of interests No competing interests are declared., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bacterial therapy is recognized as a cost-effective treatment for several diseases. However, its development is hindered by limited functionality, weak inherent therapeutic effects, and vulnerability to harsh microenvironmental conditions, leading to suboptimal treatment activity. Enhancing bacterial activity and therapeutic outcomes emerges as a pivotal challenge. Nanozymes have garnered significant attention due to their enzyme-mimic activities and high stability. They enable bacteria to mimic the functions of gene-edited bacteria expressing the same functional enzymes, thereby improving bacterial activity and therapeutic efficacy. This review delineates the therapeutic mechanisms of bacteria and nanozymes, followed by a summary of strategies for preparing bacteria/nanozyme composites. Additionally, the synergistic effects of such composites in biomedical applications such as gastrointestinal diseases and tumors are highlighted. Finally, the challenges of bacteria/nanozyme composites are discussed and propose potential solutions. This study aims to provide valuable insights to offer theoretical guidance for the advancement of nanomaterial-assisted bacterial therapy., (© 2024 Wiley‐VCH GmbH.)

Polycyclic aromatic compounds and petroleum hydrocarbons (PHs) are hazardous pollutants and seriously threaten the environment and human health. However, native microbial communities can adapt to these toxic pollutants, utilize these compounds as a carbon source, and eventually evolve to degrade these toxic contaminants. With this in mind, we isolated 26 bacterial strains from various environmental soil samples. Utilizing whole genome shotgun sequencing and analyses of these genomes revealed that they all belong to a single phylum with seven genera and sixteen species, and displayed variable genome sizes with CDS features, % GC contents, and GC skews. The analysis of genome annotation predicted genes/enzymes related to aromatic compound degradation, including the metabolism of homogentisate, salicylate and gentisate catabolism, benzoate, biphenyl, and phenylpropanoid compound degradation, and protocatechuate branch of beta-ketoadipate pathways. The majority of enzymes were found to belong to species Achromobacter pulmonis A (16%) & Achromobacter mucicolens (15%), Pseudomonas citronellolis (10%), and Comamonas thiooxydans (8%). Conversely, the highest number of hydrocarbon-degrading enzymes were found to be annotated in the species Pseudomonas citronellolis (13%), Comamonas thiooxydans (9%), Acinetobacter variabilis (7%), Pseudomonas aeruginosa, and Pseudomonas E sp002113165 (6%). These enzymes were categorized as dioxygenase, monooxygenase, hydroxylase, dehydrogenase, hydrolase, decarboxylase, aldolase, etc., and were predicted to function for benzoate, benzene, toluene, naphthalene, xylene, phthalate & terephthalate, anthranilate, protocatechuate & homoprotocatechuate, salicylate, aerobic & anaerobic gallate, and lignin subunit degradation, and catechol meta & ortho-cleavage pathways. In the future, molecular and biochemical characterization of these enzymes, together with strain assays for their capacity to degrade various pollutants, will help to improve the bioremediation process for environmental contaminations., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Bacterial biofilms pose significant challenges, from healthcare-associated infections to biofouling in industrial systems, resulting in significant health impacts and financial losses globally. Classic antimicrobial methods often fail to eradicate sessile microbial communities within biofilms, requiring innovative approaches. This review explores the structure, formation, and role of biofilms, highlighting the critical importance of exopolysaccharides in biofilm stability and resistance mechanisms. We emphasize the potential of microbial enzymatic approaches, particularly focusing on glycosidases, proteases, and deoxyribonucleases, which can disrupt biofilm matrices effectively. We also delve into the importance of enzymes such as cellobiose dehydrogenase, which disrupts biofilms by degrading polysaccharides. This enzyme is mainly sourced from Aspergillus niger and Sclerotium rolfsii, with optimized production strategies enhancing its efficacy. Additionally, we explore levan hydrolase, alginate lyase, α-amylase, protease, and lysostaphin as potent antibiofilm agents, discussing their microbial origins and production optimization strategies. These enzymes offer promising avenues for combating biofilm-related challenges in healthcare, environmental, and industrial settings. Ultimately, enzymatic strategies present environmentally friendly solutions with high potential for biofilm management and infection control., Competing Interests: Declarations. Ethics approval and consent to participate: not applicable. Consent for publication: not applicable. Conflict of interest: The authors confirm the absence of conflict of interest., (© 2024. The Author(s).)

Topoisomerases are the main enzymes capable of resolving the topological constraints imposed by DNA transactions such as transcription or replication. All bacteria possess topoisomerases of different types. Although bacteria with circular replicons should encounter similar DNA topology issues, the distribution of topoisomerases varies from one bacterium to another, suggesting polymorphic functioning. Recently, several proteins restricting, enhancing or modifying the activity of topoisomerases were discovered, opening the way to a new area of understanding DNA topology management during the bacterial cell cycle. In this review, we discuss the distribution of topoisomerases across the bacterial phylum and current knowledge on the interplay among the different topoisomerases to maintain topological homeostasis., (© 2024 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Difructose anhydrides (DFAs), distinctive cyclic disaccharides mainly naturally produced by heating (caramelization), serve as potential candidates of functional sugars that modern humans consume on a daily basis due to their remarkable physiological effects. This review explores the complex domain of specialized enzymes implicated in the metabolism of DFAs, covering the entire process from biosynthesis to degradation. We provide a detailed examination of the enzymes responsible for DFA formation and degradation, specifically those classified within the GH91, GH32, and GH172 glycoside hydrolase families. Furthermore, the evolutionary relationships among the related enzymes were systematically analyzed. Subsequently, the underlying enzymatic mechanisms that drive DFAs' metabolism were elucidated, and key insights into the intricate interplay between enzyme structure and function were unveiled. Additionally, innovative strategies for enzyme engineering were discussed, aimed at improving thermostability, enhancing catalytic activity, and altering catalytic function. Finally, the applications of the related enzymes were comprehensively summarized with a focus on their product yields, conversion rates, and methods for product purification. Here, the review presents a comprehensive investigation into enzymatic degradation and biosynthesis pathways of DFAs.

Amylases are pivotal enzymes with extensive industrial applications, including food processing, textile manufacturing, pharmaceuticals, and biofuel production. Traditional methods for enhancing amylase production in microbial strains often lack precision and efficiency. The advent of CRISPR/Cas9 technology has revolutionized genetic engineering, offering precise and targeted modifications to microbial genomes. This review explores the potential of CRISPR/Cas9 for improving amylase production, highlighting its advantages over conventional methods. This review discusses the mechanism of CRISPR/Cas9, the identification and targeting of key genes involved in amylase synthesis and regulation, and the optimization of expression systems. Additionally, current review examines case studies demonstrating successful CRISPR/Cas9 applications in various microbial hosts. The review also delves into the integration of CRISPR/Cas9 in wastewater treatment, where genetically engineered amylolytic strains enhance the degradation of complex organic pollutants. Despite the promising prospects, challenges such as off-target effects and regulatory considerations remain. This review provides a comprehensive overview of the current advancements, challenges, and future directions in the application of CRISPR/Cas9 technology for amylase production and environmental biotechnology., Competing Interests: Declarations. Conflicts of interests: The authors declare that they have no conflicts of interest or competing interests related to this publication. Ethics Approval: Not applicable. This study is a review of existing literature and does not involve human or animal subjects requiring ethical approval. Consent to Participate: Not applicable. This review paper is a synthesis of existing research and does not involve human participants, animal subjects, or primary data collection requiring explicit consent for participation. Consent for Publication: Not applicable. As this is a review paper compiling data from previously published studies, no additional permissions for publication are required beyond citing the original sources. All included material is publicly available and used under the principles of academic integrity and proper attribution., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Species-specific enzymes provide a substantial boost to the precision and selectivity of identifying dairy products contaminated with foodborne pathogens, due to their specificity for target organisms. In this study, we developed cobalt oxyhydroxide nanosheets (CoOOH NSs) for a dual-mode biosensor capable of detecting β-galactosidase (β-Gal)-positive bacteria in milk and milk powder. The sensor exploits the oxidase-mimicking activity of CoOOH NSs, where β-Gal converts the substrate β-D-galactopyranoside to p-aminophenol, reducing CoOOH NSs to Co 2+ and inhibiting the formation of the blue product from 3,3',5,5'-tetramethylben-zidine. Sensitivity was enhanced through membrane filtration and β-Gal induction by isopropyl β-D-thiogalactoside. The assay achieved a detection limit of 5 cfu mL -1 and demonstrated recoveries (90.7 % to 103 %) and relative standard deviations <5.7 % in milk and milk powder samples. These findings underscore the potential of the sensor for detecting β-Gal-positive bacteria in dairy products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Submerged plants can thrive entirely underwater, playing a crucial role in maintaining water quality, supporting aquatic organisms, and enhancing sediment stability. However, they face multiple challenges, including reduced light availability, fluctuating water conditions, and limited nutrient access. Despite these stresses, submerged plants demonstrate remarkable resilience through physiological and biochemical adaptations. Additionally, their interactions with microbial communities are increasingly recognized as pivotal in mitigating these environmental stresses. Understanding the diversity of these microbial communities is crucial for comprehending the complex interactions between submerged plants and their environments. This research aims to identify and screen microbes from submerged plant samples capable of producing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and to explore microbial diversity through metagenomic analysis. Microbes were isolated and screened for ACC deaminase production, and metagenomic techniques, including co-occurrence network analysis, were used to examine microbial diversity and interactions within the communities. ACC deaminase-producing microbes can significantly enhance plant metabolism under stress conditions. The identification of the culturable bacteria revealed that most of these microbes belong to the genera Pseudomonas , Bacillus , and Acinetobacter . A total of 177 microbial strains were cultured, with molecular identification revealing 79 reductant, 86 non-reductant, and 12 uncultured strains. Among 162 samples screened for ACC deaminase activity, 50 tested positive. To further understand microbial dynamics, samples were collected from both natural sources and artificial pond reservoirs to assess the impact of the location on flood-associated microbiomes in submerged plants. Metagenomic analysis was conducted on both the epiphytic and endophytic samples. By exploring the overall composition and dynamics of microbial communities associated with submerged plants, this research seeks to deepen our understanding of plant-microbe interactions in aquatic environments. The microbial screening helped to identify the diverse microbes associated with ACC deaminase activity in submerged plants and amplicon sequencing analysis paved the way towards identifying the impact of the location in shaping the microbiome and the diversity associated with endophytic and epiphytic microbes. Co-occurrence network analysis further highlighted the intricate interactions within these microbial communities. Notably, ACC deaminase activity was observed in plant-associated microbes across different locations, with distinct variations between epiphytic and endophytic populations as identified through co-occurrence network analysis.

Recent studies show that biodegradable microplastics (BMPs) could increase soil CO 2 emission, but whether altered carbon emission results from modified soil organic matter (SOM) decomposition remains underexplored. In this study, the effect and mechanisms of BMPs on CO 2 emission from soil were investigated, using poly(butylene adipate-co-terephthalate) (PBAT, the main component of agricultural film) as an example. Considering that straw returning is a common agronomic measure which may interact with microplastics through affecting microbial activity, both soils with and without wheat straw were included. After 120 d, 1 % (w/w) PBAT BMPs ificantly increased cumulative CO 2 emission by 1605.6 and 1827.7 mg C kg -1 in soils without and with straw, respectively. Cracks occurred on the surface of microplastics, indicating that CO 2 was partly originated from plastic degradation. Soil dissolved organic matter (DOM) content, carbon degradation gene abundance (such as abfA, xylA and manB for hemicellulose, mnp, glx and lig for lignin, and chiA for chitin) and enzyme activities increased, which significantly positively correlated with CO 2 emission rate (p < 0.05), suggesting that PBAT enhanced carbon emission by stimulating the decomposition of SOM (and possibly the newly added straw) via co-metabolism and nitrogen mining. This is supported by DOM molecular composition analysis which also demonstrated stimulated turnover of carbohydrates, amino sugars and lignin following PBAT addition. The findings highlight the potential of BMPs to affect SOM stability and carbon emission., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Per- and polyfluorinated chemicals (PFAS) are of rising concern due to environmental persistence and emerging evidence of health risks to humans. Environmental persistence is largely attributed to a failure of microbes to degrade PFAS. PFAS recalcitrance has been proposed to result from chemistry, specifically C-F bond strength, or biology, largely negative selection from fluoride toxicity. Given natural evolution has many hurdles, this review advocates for a strategy of laboratory engineering and evolution. Enzymes identified to participate in defluorination reactions have been discovered in all Enzyme Commission classes, providing a palette for metabolic engineering. In vivo PFAS biodegradation will require multiple types of reactions and powerful fluoride mitigation mechanisms to act in concert. The necessary steps are to: (1) engineer bacteria that survive very high, unnatural levels of fluoride, (2) design, evolve, and screen for enzymes that cleave C-F bonds in a broader array of substrates, and (3) create overall physiological conditions that make for positive selective pressure with PFAS substrates., (© 2024 The Author(s).)

Lignin is an aromatic biopolymer found in plant cell walls and is the most abundant source of renewable aromatic carbon in the biosphere. Hence there is considerable interest in the conversion of lignin, either derived from agricultural waste or produced as a byproduct of pulp/paper manufacture, into high-value chemicals. Although lignin is rather inert, due to the presence of ether C-O and C-C linkages, several microbes are able to degrade lignin. This review will introduce these microbes and the enzymes that they use to degrade lignin and will describe recent studies on metabolic engineering that can generate high-value chemicals from lignin bioconversion. Catabolic pathways for degradation of lignin fragments will be introduced, and case studies where these pathways have been engineered by gene knockout/insertion to generate bioproducts that are of interest as monomers for bioplastic synthesis or aroma chemicals will be described. Life cycle analysis of lignin bioconversion processes is discussed.

This work describes determining urea in milk samples using a multicommuted approach with a urease enzyme immobilized in bacterial cellulose and solid MOF as a colorimetric reagent. The Cu(2+)-MOF was characterized by FTIR spectroscopy, XRD, and SEM. The urea quantification was based on the urea hydrolysis reaction catalyzed by urease and reacted with Cu(2+)-MOF forming [Cu(NH 3 ) 4 ] 2+ , monitored at 450 nm. Linear responses were obtained from 1.0 to 50.0 mg dL -1 urea (R = 0.9959, n = 11), detection and quantitation limits of 0.082 mg dL -1 and 0.272 mg dL -1 respectively, analytical frequency of 8 determinations per hour, 0.8 mL sample solution consumption. Potential interfering studies have shown the selectivity of the proposed method. Addition and recovery tests were performed obtaining variation from 90 to 103%. Applying the F-test and t-test, the results showed no significant difference at the 95% confidence level Comparing the proposed and the reference method., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence this work., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The evolution, survival, and adaptation of microbes are consequences of gene duplication, acquisition, and divergence in response to environmental challenges. In this context, enzymes play a central role in the evolution of organisms, because they are fundamental in cell metabolism. Here, we analyzed the enzymatic repertoire in 6,467 microbial genomes, including their abundances, and their associations with metabolic maps. We found that the enzymes follow a power-law distribution, in relation to the genome sizes. Therefore, we evaluated the total proportion enzymatic classes in relation to the genomes, identifying a descending-order proportion: transferases (EC:2.-), hydrolases (EC:3.-), oxidoreductases (EC:1.-), ligases (EC:6.-), lyases (EC:4.-), isomerases (EC:5.-), and translocases (EC:7-.). In addition, we identified a preferential use of enzymatic classes in metabolism pathways for xenobiotics, cofactors and vitamins, carbohydrates, amino acids, glycans, and energy. Therefore, this analysis provides clues about the functional constraints associated with the enzymatic repertoire of functions in Bacteria and Archaea., Competing Interests: Declarations. Dirección General de Asuntos del Personal Académico-Universidad Nacional Autónoma de México (IN-220523), Consejo Nacional de Humanidades, Ciencias y Tecnologías (320012). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., (© 2024. The Author(s).)

Enzymes play a crucial role in various industrial sectors. These biocatalysts not only ensure sustainability and safety but also enhance process efficiency through their unique specificity. Lipases possess versatility as biocatalysts and find utilization in diverse bioconversion reactions. Presently, microbial lipases are gaining significant focus owing to the rapid progress in enzyme technology and their widespread implementation in multiple industrial procedures. This updated review presents new knowledge about various origins of microbial lipases, such as fungi, bacteria, and yeast. It highlights both the traditional and modern purification methods, including precipitation and chromatographic separation, the immunopurification technique, the reversed micellar system, the aqueous two-phase system (ATPS), and aqueous two-phase flotation (ATPF), moreover, delves into the diverse applications of microbial lipases across several industries, such as food, vitamin esters, textile, detergent, biodiesel, and bioremediation. Furthermore, the present research unveils the obstacles encountered in employing lipase, the patterns observed in lipase engineering, and the application of CRISPR/Cas genome editing technology for altering the genes responsible for lipase production. Additionally, the immobilization of microorganisms' lipases onto various carriers also contributes to enhancing the effectiveness and efficiencies of lipases in terms of their catalytic activities. This is achieved by boosting their resilience to heat and ionic conditions (such as inorganic solvents, high-level pH, and temperature). The process also facilitates the ease of recycling them and enables a more concentrated deposition of the enzyme onto the supporting material. Consequently, these characteristics have demonstrated their suitability for application as biocatalysts in diverse industries., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The flavor alterations in bighead carp subjected to varying storage temperatures and the underlying metabolic mechanism were elucidated. Analysis of volatile flavor compounds, electronic nose, free amino acids, ATP-related compounds, and sensory evaluations uncovered a progressive flavor deterioration during storage, especially at 25 °C. Metabolomics-based flavor relating component profiling analysis showed that free fatty acids formed various fatty aldehydes including (E, E)-2,4-heptadienal and nonanal under lipoxygenase catalysis. Alcohol dehydrogenase and alcohol acyltransferases were intimately involved in alcohol and ester generation, while alkaline phosphatase, 5'-nucleotidase, and acid phosphatase were closely associated with IMP, Hx, and HxR conversion, respectively. Aeromonas, Serratia, Lactococcus, Pseudomonas, and Peptostreptococcus notably influenced flavor metabolism and enzyme activities. The metabolism disparities of valine, leucine, isoleucine, lysine, and α-linolenic acid could be the primary factors contributing to flavor metabolism distinctions. This study offers novel insights into the flavor change mechanisms and potential regulation strategies of bighead carp during storage., Competing Interests: Declaration of competing interest The authors declare that they do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Plant pathogens deploy specialized proteins to aid disease progression, some of these proteins function in the apoplast and constitute proteases. Extracellular virulence proteases have been described to play roles in plant cell wall manipulation and immune evasion. In this review, we discuss the evidence for the hypothesized virulence functions of bacterial, fungal, and oomycete extracellular proteases. We highlight the contrast between the low number of elucidated functions for these proteins and the size of extracellular protease repertoires among pathogens. Finally, we suggest that the refinement of in planta 'omics' approaches, combined with recent advances in modeling and mechanism-based methods for trapping substrates finally make it possible to address this knowledge gap., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Topoisomerases are essential, ubiquitous enzymes that break and rejoin the DNA strand to control supercoiling. Because topoisomerases are DNA scissors, these enzymes are highly regulated to avoid excessive DNA cleavage, a vulnerability exploited by many antibiotics. Topoisomerase activity must be co-ordinated in time and space with transcription, replication, and cell division or else these processes stall, leading to genome loss. Recent work in Escherichia coli has revealed that topoisomerases do not act alone. Most topoisomerases interact with the essential process that they promote, a coupling that may stimulate topoisomerase activity precisely when and where cleavage is required. Surprisingly, in E. coli and most other bacteria, gyrase is not apparently regulated in this manner. We review how each E. coli topoisomerase is regulated, propose possible solutions to 'the gyrase problem', and conclude by highlighting how this regulation may present opportunities for antimicrobial development., Competing Interests: Declaration of Competing Interest Sora Kim reports financial support was provided by Helen Hay Whitney Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Ubiquitin (Ub) and ubiquitin-like (UbL) modifications are critical regulators of multiple cellular processes in eukaryotes. These modifications are dynamically controlled by proteases that balance conjugation and deconjugation. In eukaryotes, these proteases include deubiquitinases (DUBs), mostly belonging to the CA-clan of cysteine proteases, and ubiquitin-like proteases (ULPs), belonging to the CE-clan proteases. Intriguingly, infectious bacteria exploit the CE-clan protease fold to generate deubiquitinating activities to disarm the immune system and degradation defenses of the host during infection. In this review, we explore the substrate preferences encoded within the CE-clan proteases and the structural determinants in the protease fold behind its selectivity, in particular those from infectious bacteria and viruses. Understanding this protease family provides crucial insights into the molecular mechanisms underlying infection and transmission of pathogenic organisms., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

During storage, the proteins of gazami crab (Portunus trituberculatus) are prone to hydrolysis into amino acids and biogenic amines, in which enzymes play a critical role. However, studies exploring spoilage mechanisms from the perspective of enzymes are limited. This study identified 84 endogenous and 52 microbial-derived proteolytic enzymes and peptidases by proteomics and metagenomics. There are 7 endogenous amino acid deaminases, primarily degrade glutamate and aspartate. Additionally, 25 amino acid deaminases of microbial origin were identified, which mainly degrade serine. The formation of biogenic amines involved 14 enzymes, all of which were microbial in origin, primarily synthesizing putrescine from arginine. The main microbial contributors to these enzymes were Photobacterium, Vibrio, and Aliivibrio, accounting for 63.87 %, 15.51 %, and 8.69 % at the end of refrigeration, respectively. This study provides insights into the mechanisms of quality deterioration in gazami crab during refrigeration, from the perspectives of metabolic enzymes and microbial activity., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

α-amylases are vital biocatalysts that constitute a billion-dollar industry with a substantial and enduring global demand. Amylases hydrolyze the α-1,4-glycosidic linkages in starch polymers to generate maltose and malto-oligosaccharides subunits. Amylases are key enzymes that have promising applications in various industrial processes ranging from pharmaceutical, pulp and paper, textile food industries to bioremediation and biofuel sectors. Microbial enzymes have been widely used in industrial applications owing to their ease of availability, cost-effectiveness and better stability at extreme temperatures and pH. α-amylases derived from distinct microbial origins exhibit diverse characteristics, which make them suitable for specific applications. The routine application of immobilized enzymes has become a standard practice in the production of numerous industrial products across the pharmaceutical, chemical, and food industries. This review details the structural makeup of microbial α-amylase to understand its thermodynamic characteristics, aiming to identify key areas that could be targeted for improving the thermostability, pH tolerance and catalytic activity of α-amylase through various immobilization techniques or specific enzyme engineering methods. Additionally, the review briefly explores the enzyme production strategies, potential sources of α-amylases, and use of cost-effective and sustainable raw materials for enzyme production to obtain α-amylases with unconventional applications in various industrial sectors. Major hurdles, challenges and future prospects involving microbial α-amylases has been briefly discussed by considering its diverse applications in industrial bioprocessing., Competing Interests: Declaration of competing interest The authors have no financial or non-financial competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Xiaoqu , a pivotal starter in baijiu fermentation, provides the most microflora and enzymes to initiate and maintain baijiu brewing. This study aims to explore the differences in microbiota and enzyme activities among Xiaoqu samples from seven provinces in southern China using high-throughput sequencing, plate isolation, and activity detection. The analyses revealed significant differences in bacterial and fungal communities across the samples. A total of 22 bacterial species and 17 target fungal species were isolated and identified. Predominant bacteria included Bacillus ( Bacillus subtilis ) and lactic acid bacteria (LABs), while the fungal communities were primarily composed of yeasts ( Saccharomyces cerevisiae ) and various molds. The activities of α-amylase and glucoamylase varied significantly among the samples, and samples from HN1 and GZ2 exhibited the highest activities. Correlation analyses highlighted the pivotal role of LABs in maintaining acidity and the importance of molds and yeasts in the saccharification and fermentation processes. These findings shed light on the microbial composition and diversity of Xiaoqu and the critical role of microbes in baijiu production. Moreover, they suggested potential microbial resources for developing artificial Xiaoqu via synthetic microbial community in the future, enhancing baijiu fermentation efficiency and overall product quality.

Cobamides (Cbas) are cobalt-containing cyclic tetrapyrroles used by cells from all domains of life as co-catalyst of diverse reactions. There are several structural features that distinguish Cbas from one another. The most relevant of those features discussed in this review is the lower ligand, which is the nucleobase of a ribotide located in the lower face of the cyclic tetrapyrrole ring. The above-mentioned ribotide is known as the nucleotide loop, which is attached to the ring by a short linker. In Cbas, the nucleobase of the ribotide can be benzimidazole or derivatives of it, purine or derivatives of it, or phenolic compounds. Given the importance of Cbas in prokaryotic metabolism, it is not surprising that prokaryotes have evolved enzymes that cleave part or the entire nucleotide loop. This function is advantageous when Cbas contain nucleobases that somehow interfere with the function of Cba-dependent enzymes in the organism. After cleavage, Cbas are rebuilt via the nucleotide loop assembly (NLA) pathway, which includes enzymes that activate the nucleobase and the ring intermediate, followed by condensation of activated intermediates and a final dephosphorylation reaction. This exchange of nucleobases is known as Cba remodeling. The NLA pathway is used to salvage Cba precursors from the environment., Competing Interests: The authors declare no conflict of interest.

In eggs, there are several components: eggshell (ES), eggshell membrane (ESM), egg white (EW), and egg yolk (EY). Many modification methods exist, such as thermal treatment, high pressure, freeze-thaw cycles, ultrasonic treatment, ozonation, phosphorylation, and acylation, all aimed at improving the functional properties of EW and EY. Additionally, microorganism and enzyme modifications have proven effective in enhancing the functional properties of EW and EY. ES and ESM are unique components of eggs. The eggshell is rich in calcium carbonate, while the eggshell membrane is rich in protein. The effective utilization of ES and ESM can help promote economic income in the poultry industry and benefit the environment. Research on the modification of ES and ESM has shown that microorganisms and enzymes have the potential to improve their functional properties. After modification, egg components can be utilized in the production of egg-based and other food products for improved performance. Furthermore, enzyme modification of egg components can produce bioactive peptides, which have the potential to treat specific diseases and may even be used in the biomedical field. This review primarily focuses on the effects of microorganisms and enzymes on the modification of egg components and summarizes the roles of microbial and enzymatic modifications in this context.

The single-step methioninase-mediated degradation of methionine (as a sulfur containing amino acid) is a reaction at the interface of carbon, nitrogen, sulfur, and methane metabolism in microbes. This enzyme also has therapeutic application due to its role in starving auxotrophic cancer cells. Applying our refined in silico screening pipeline on 33,469 publicly available genome assemblies and 1878 metagenome assembled genomes/single-cell amplified genomes from brackish waters of the Caspian Sea and the Fennoscandian Shield deep groundwater resulted in recovering 1845 methioninases. The majority of recovered methioninases belong to representatives of phyla Proteobacteria (50%), Firmicutes (29%), and Firmicutes&#95;A (13%). Prevalence of methioninase among anaerobic microbes and in the anoxic deep groundwater together with the relevance of its products for energy conservation in anaerobic metabolism highlights such environments as desirable targets for screening novel methioninases and resolving its contribution to microbial metabolism and interactions. Among archaea, majority of detected methioninases are from representatives of Methanosarcina that are able to use methanethiol, the sulfur containing product from methionine degradation, as a precursor for methanogenesis. Branching just outside these archaeal methioninases in the phylogenetic tree, we recovered three methioninases belonging to representatives of Patescibacteria reconstructed from deep groundwater metagenomes. We hypothesize that methioninase in Patescibacteria could contribute to their syntrophic interactions where their methanogenic partners/hosts benefit from the produced 2-oxobutyrate and methanethiol. Our results underscore the significance of accounting for specific ecological niche in screening for enzyme variates with desired characteristics. Finally, complementing of our findings with experimental validation of methioninase activity confirms the potential of our in silico screening in clarifying the peculiar ecological role of methioninase in anoxic environments., Competing Interests: Declarations Conflict of Interest The authors declare no competing interests., (© 2024. The Author(s).)

Nitrogen is often a limiting nutrient for microbial communities and plants in glacier forefields. Nitrogen-fixing microorganisms (diazotrophs) play an important role in providing bioavailable nitrogen, with their composition determining the nitrogen-fixating capacities. This study investigates the spatial and temporal dynamics of diazotrophs in the forefields of three Tibetan glaciers: Qiangyong, Kuoqionggangri, and Longxiazailongba. We collected soil samples from recently deglaciated barren grounds, and also along an ecosystem succession transect at Kuoqionggangri glacier, encompassing barren ground, herb steppe, legume steppe, and alpine meadow ecosystems. Our finding revealed abundant and diverse diazotrophs in the recently deglaciated barren ground. They are taxonomically affiliated with anaerobic Bradyrhizobium, Desulfobulbus, and Pelobacter, which may be relics from subglacial sediments. The vegetated soils (herb steppe, legume steppe, and alpine meadow) were dominated by phototrophic Nostoc and Anabaena, as well as symbiotic Sinorhizobium. Soil physicochemical parameters, such as soil organic carbon, pH, and nitrate ion, significantly influenced diazotroph community structure. This study highlights the critical role of diazotrophs in mitigating nitrogen limitation during early ecosystem development in glacier forefields. Understanding the distribution and ecological drivers of diazotrophs in these rapidly changing environments provides insights into biogeochemical cycling and ecosystem resilience under climate change., Competing Interests: Declarations Conflict of Interest The authors declare no competing interests., (© 2024. The Author(s).)

Background: Wood-feeding termites have been employed as sources of novel and highly efficient lignocellulolytic enzymes due to their ability to degrade lignocellulose efficiently. As a higher wood-feeding termite, Globitermes sulphureus (Blattaria: Termitidae) plays a crucial role as a decomposer in regions such as Vietnam, Singapore, Myanmar, and Yunnan, China. However, the diversity of its gut microbiome and carbohydrate-active enzymes (CAZymes) remains unexplored. Here, we analyzed the diversity of hindgut microbial communities and CAZymes in a higher wood-feeding termite, G. sulphureus, and a lower wood-feeding termite, Coptotermes formosanus (Blattaria: Rhinotermitidae)., Results: 16S rRNA sequencing revealed that Spirochaetota, Firmicutes, and Fibrobacterota were the dominant microbiota in the hindgut of the two termite species. At the phylum level, the relative abundances of Proteobacteria and Bacteroidota were significantly greater in the hindgut of C. formosanus than in G. sulphureus. At the genus level, the relative abundances of Candidatus&#95;Azobacteroides and Escherichia-Shigella were significantly lower in the hindgut of G. sulphureus than in C. formosanus. Metagenomic analysis revealed that glycoside hydrolases (GHs) with cellulases and hemicellulases functions were not significantly different between G. sulphureus and C. formosanus. Interestingly, the cellulases in G. sulphureus were mainly GH5&#95;2, GH5&#95;4, GH6, GH9, and GH45, while the hemicellulases were mainly GH11, GH8, GH10, GH11, GH26, and GH53. In C. formosanus, the cellulases were mainly GH6 and GH9, and the hemicellulases were mainly GH5&#95;7, GH5&#95;21, GH10, GH12, and GH53. In addition, β-glucosidase, exo-β-1,4-glucanase, and endo-β-1,4-glucanase activities did not differ significantly between the two termite species, while xylanase activity was higher in G. sulphureus than in C. formosanus. The bacteria encoding GHs in G. sulphureus were mainly Firmicutes, Fibrobacterota, and Proteobacteria, whereas Bacteroidota and Spirochaetota were the main bacteria encoding GHs in C. formosanus., Conclusions: Our findings characterized the microbial composition and differences in the hindgut microbiota of G. sulphureus and C. formosanus. Compared to C. formosanus, G. sulphureus is enriched in genes encoding for hemicellulase and debranching enzymes. It also highlights the rich diversity of GHs in the hindgut microbiota of G. sulphureus, including the GH5 subfamily, GH6, and GH48, with the GH6 and GH48 not previously reported in other higher termites. These results strengthen the understanding of the diversity of termite gut microbiota and CAZymes., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Metalloenzymes play central roles in the anaerobic metabolism of human gut microbes. They facilitate redox and radical-based chemistry that enables microbial degradation and modification of various endogenous, dietary, and xenobiotic nutrients in the anoxic gut environment. In this review, we highlight major families of iron-sulfur (Fe-S) cluster-dependent enzymes and molybdenum cofactor-containing enzymes used by human gut microbes. We describe the metabolic functions of 2-hydroxyacyl-CoA dehydratases, glycyl radical enzyme activating enzymes, Fe-S cluster-dependent flavoenzymes, U32 oxidases, and molybdenum-dependent reductases and catechol dehydroxylases in the human gut microbiota. We demonstrate the widespread distribution and prevalence of these metalloenzyme families across 5000 human gut microbial genomes. Lastly, we discuss opportunities for metalloenzyme discovery in the human gut microbiota to reveal new chemistry and biology in this important community., (© The Author(s) 2024. Published by Oxford University Press.)

Polyamide 4 (PA4) is expected to solve the issue of marine plastic pollution due to its excellent mechanical properties and biodegradability. In this study, to reveal the mechanism of PA4 biodegradation in the marine environment, we isolated 5 strains of PA4-degrading bacteria belonging to Aliiglaciecola, Dasania, and Pseudophaeobacter from a marine environment. The isolated 5 strains are novel PA4-degrading bacteria that are phylogenetically distinct from those isolated in previous studies. In addition, we compared the PA4-degrading activities and structures of the PA4-degrading enzymes secreted by the 5 strains and PA4-degrading strains isolated in our previous study. The PA4-degrading activity in the supernatant of the cultivation solutions differed among the strains. Native-PAGE and zymography using a polyacrylamide gel containing a PA4 emulsion demonstrated that PA4-degrading enzymes are classified into no less than three types of structures. These results suggested that marine PA4-degrading bacteria have multiple PA4-degrading enzymes. Our findings will contribute to a better understanding of the microbial degradation of PA4 in the marine environment.

Termites digest wood using Carbohydrate-Active Enzymes (CAZymes) produced by gut bacteria with whom they have cospeciated at geological timescales. Whether CAZymes were encoded in the genomes of their ancestor's gut bacteria and transmitted to modern termites or acquired more recently from bacteria not associated with termites is unclear. We used gut metagenomes from 195 termites and one Cryptocercus, the sister group of termites, to investigate the evolution of termite gut bacterial CAZymes. We found 420 termite-specific clusters in 81 bacterial CAZyme gene trees, including 404 clusters showing strong cophylogenetic patterns with termites. Of the 420 clusters, 131 included at least one bacterial CAZyme sequence associated with Cryptocercus or Mastotermes, the sister group of all other termites. Our results suggest many bacterial CAZymes have been encoded in the genomes of termite gut bacteria since termite origin, indicating termites rely upon many bacterial CAZymes endemic to their guts to digest wood., (© 2024. The Author(s).)

Aims: This study continues our previous investigation of the intrinsic degradation of phosphogypsum (PG) by indigenous microorganisms on amending adequate nutrients. We aim to unravel the intricate mechanisms involved in PG biotransformation by a bacterial consortium., Methods and Results: We isolated and characterized seven multi-metal-resistant bacterial strains from a nutrient-amended PG-contaminated microcosm and identified them through 16S rRNA gene sequencing. Primarily aerobic, Gram-positive chemolithotrophs, these strains demonstrated significant heavy metal uptake and PG degradation potential. Further analysis revealed that all strains produced carbonic anhydrase (CA), while six also produced urease, which may facilitate microbial-induced carbonate precipitation. Microstructural and elemental analysis using scanning electron microscopy-energy-dispersive X-ray and X-Ray Diffraction (XRD) confirmed the PG bio-transformation, indicating substantial increases in carbonate concentrations and reductions in sulfate levels., Conclusions: The consortium, composed of seven urease- and CA-producing bacterial strains, effectively degraded PG, transforming it from an acidic to an alkaline state and significantly enhancing CO2 sequestration., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Classical Zn 2+ -dependent deac(et)ylases play fundamental regulatory roles in life and are well characterized in eukaryotes regarding their structures, substrates and physiological roles. In bacteria, however, classical deacylases are less well understood. We construct a Generalized Profile (GP) and identify thousands of uncharacterized classical deacylases in bacteria, which are grouped into five clusters. Systematic structural and functional characterization of representative enzymes from each cluster reveal high functional diversity, including polyamine deacylases and protein deacylases with various acyl-chain type preferences. These data are supported by multiple crystal structures of enzymes from different clusters. Through this extensive analysis, we define the structural requirements of substrate selectivity, and discovered bacterial de-D-/L-lactylases and long-chain deacylases. Importantly, bacterial deacylases are inhibited by archetypal HDAC inhibitors, as supported by co-crystal structures with the inhibitors SAHA and TSA, and setting the ground for drug repurposing strategies to fight bacterial infections. Thus, we provide a systematic structure-function analysis of classical deacylases in bacteria and reveal the basis of substrate specificity, acyl-chain preference and inhibition., (© 2024. The Author(s).)

The production of the sugars fructose and lactulose from lactose using the enzymes β-galactosidase and glucose isomerase immobilized on bacterial cellulose (BC) membranes has been investigated. Lactose is hydrolyzed by β-galactosidase at 30 °C to glucose and galactose at a high conversion rate, while at the same temperature, glucose isomerase is not effective in converting the produced glucose to fructose. The rate of the isomerization reaction of glucose to fructose at 70 °C has been studied. Two types of enzyme immobilization were investigated: immobilization in one stage and immobilization in two stages. The results showed that BC membrane increased three-fold the yield and the reaction rate of fructose and lactulose production from lactose. The noteworthy enhancement of BC membranes' impact on the isomerization reaction by immobilized enzymes grants permission for a novel research avenue within the context of white biotechnology development. Additionally, this effect amplifies the role of BC in sustainability and the circular economy., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kanellaki Maria reports financial support was provided by Epanek2014–2020 Operational Programme., (Copyright © 2024. Published by Elsevier Ltd.)

Prokaryotic anti-phage immune systems use TIR and cGAS-like enzymes to produce 1''-3'-glycocyclic ADP-ribose (1''-3'-gcADPR) and cyclic dinucleotide (CDN) and cyclic trinucleotide (CTN) signalling molecules, respectively, which limit phage replication 1-3 . However, how phages neutralize these distinct and common systems is largely unclear. Here we show that the Thoeris anti-defence proteins Tad1 4 and Tad2 5 both achieve anti-cyclic-oligonucleotide-based anti-phage signalling system (anti-CBASS) activity by simultaneously sequestering CBASS cyclic oligonucleotides. Apart from binding to the Thoeris signals 1''-3'-gcADPR and 1''-2'-gcADPR, Tad1 also binds to numerous CBASS CDNs and CTNs with high affinity, inhibiting CBASS systems that use these molecules in vivo and in vitro. The hexameric Tad1 has six binding sites for CDNs or gcADPR, which are independent of the two high-affinity binding sites for CTNs. Tad2 forms a tetramer that also sequesters various CDNs in addition to gcADPR molecules, using distinct binding sites to simultaneously bind to these signals. Thus, Tad1 and Tad2 are both two-pronged inhibitors that, alongside anti-CBASS protein 2 (Acb2 6-8 ), establish a paradigm of phage proteins that use distinct binding sites to flexibly sequester a considerable breadth of cyclic nucleotides., Competing Interests: Competing interests: J.B.-D. is a scientific advisory board member of SNIPR Biome and Excision Biotherapeutics, a consultant to LeapFrog Bio and BiomX, and a scientific advisory board member and co-founder of Acrigen Biosciences and ePhective Therapeutics. The J.B.-D. laboratory received prior research support from Felix Biotechnology., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)