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The production of fermentable sugars from lignocellulosic biomass is achieved by the synergistic action of a group of enzymes called cellulases. Cellulose is a long chain of chemically linked glucoses by β-1,4 bonds. The enzyme β-1,4-endoglucanase is the first cellulase involved in the degradation, breaking the bond of the amorphous regions. A β-1,4-endoglucanase enzyme with high activity was obtained from a Bacillus subtilis strain isolated from wastewater of a pulp and paper mill. Sequencing and bioinformatic analysis showed that the gene amplified by PCR consisting of 1407 nucleotides and coding for a β-1,4-endoglucanase enzyme of approximately 55 kDa. The open reading frame (ORF) encoding the mature endoglucanase (eglS) was successfully inserted in a modified cloning plasmid (pITD03) and into the pYD1 plasmid used for its expression in yeast. Carboxymethylcellulose (CMC) plate assay, SDS-PAGE, and zymogram confirmed the production and secretion by the transformed E. coli BL21-SI strain of a 39 kDa β-1,4-endoglucanase consistent with the catalytic domain without the cellulose-binding module (CBM). The results showed that the truncated β-1,4-endoglucanase had higher activity and stability., Competing Interests: Declaration of competing interest The author(s) declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Dipicolinic acid (DPA) is an important chemical marker for the detection of bacterial spores. In this study, complexes of lanthanide series elements such as erbium, europium, neodymium, and terbium were prepared with pyrocatechol violet and effectively immobilized the pyrocatechol violet (PV)-metal complex on a filter paper using polyvinyl alcohol. These filter paper strips were employed for the onsite detection of bacterial spores. The test filter papers were evaluated quantitatively with different concentrations of DPA and spores of various bacteria. Among the four lanthanide ions, erbium displayed better sensitivity than the other ions. The limit of detection of this test for DPA was 60 μM and 5 × 10(6) spores. The effect of other non-spore-forming bacteria and interfering chemicals on the test strips was also evaluated. The non-spore-forming bacteria did not have considerable effect on the test strip whereas chemicals such as EDTA had significant effects on the test results. The present test is rapid and robust, capable of providing timely results for better judgement to save resources on unnecessary decontamination procedures during false alarms.

Although significant technological advancements have been made in the development of analytical biosensor chips for detecting bacterial strains (E. coli, S. Mutans and B. Subtilis), critical requirements i.e. limit of detection (LOD), fast time of response, ultra-sensitivity with high reproducibility and good shelf-life with robust sensing capability have yet to be met within a single sensor chip. In order to achieve these criteria, we present an electrically-receptive thermally-responsive (ER-TR) sensor chip comprised of simple filter paper used as substrate coated with composite of poly(N-isopropylacrylamide) polymer (PNIPAm) - graphene nanoplatelet (GR) followed by evaporation of Au electrodes for capturing both Gram-positive (S. mutans and B. subtilis) and Gram-negative (E. coli) bacterial cells in real-time. Autoclave water, tap water, lake water and milk samples were tested with ER-TR chip with and without bacterial strains at varying concentration range 10 1 -10 5 cells/mL. The sensor was integrated with in-house built printed circuit board (PCB) to transmit/receive electrical signals. The interaction of E. coli, S. mutans and B. subtilis cells with fibers of PNIPAm-GR resulted in a change of electrical resistance and the readout was monitored wirelessly in real-time using MATLAB algorithm. Finally, prepared ER-TR chip exhibited the reproducibility of 85-97% with shelf-life of up to four weeks after testing with lake water sample., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A new indole alkaloid, cigarindole A (1), together with five known indole alkaloids (2–6) were isolated from cigar tobacco. Their structures were elucidated using spectroscopic methods, including extensive 1D and 2D NMR techniques. Interestingly, compound 1 exhibited high antibacterial activity with bacteriostatic diameter within the range 20.6–26.7 mm against five pathogenic strains (Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae, Bacillus subtilis, and Proteus spp). Compounds 2–6 exhibited obvious antibacterial activity with bacteriostatic diameter within the range 11.9–21.8 mm. In addition, compound 1 also the potential value for use as an antiseptic agent for cigarette tipping paper. [ABSTRACT FROM AUTHOR]

Laccases (EC 1.10.3.2) are considered one of the most prominent multicopper enzymes that exhibit the inherent properties of oxidizing a range of phenolic substrates. Mostly, reported laccases have been isolated from the plants and fungi species, whereas bacterial laccases are yet to be explored. Bacterial laccases have numerous distinctive properties over fungal laccases, including stability at high temperatures and high pH. This study includes the isolation of bacteria through the soil sample collected from the paper and pulp industry; the highest laccase-producing bacteria was identified as Bhargavaea bejingensis, using 16S rRNA gene sequencing. The extracellular and intracellular activities after 24 h incubation were 1.41 U/mL and 4.95 U/mL, respectively. The laccase-encoding gene of the bacteria was sequenced; moreover, the in vitro translated protein was bioinformatically characterized and asserted that the laccase produced by the bacteria Bhargavaea bejingensis was structurally and sequentially homologous to the CotA protein of Bacillus subtilis. The enzyme laccase produced from B. bejingensis was classified as three-domain laccase with several copper-binding residues, where a few crucial copper-binding residues of the laccase enzyme were also predicted. [ABSTRACT FROM AUTHOR]

Cellulase (mainly endoglucanase, EG) has been used in pulp modification for improving paper quality through environmentally friendly process. But low activity in alkaline pH and high filter paper activity (FPA) were still obstacles for extending the cellulase application in papermaking industry. In the study, an alkali-tolerant EG gene of Bacillus subtilis Y106 was homologous over-expressed for obtaining suitable enzyme used in pulp modification. The engineering strain could produce the crude enzyme with more alkali-tolerant EG and little PFA. Potential of the crude enzyme in modification of different pulps were investigated. It was found that the enzyme could be used for improving drainage and strength properties of pulps from softwood, hardwood and non-wood materials, especially non-wood pulp such as wheat straw pulp. The underlying mechanisms of pulp modification and different effects on various types of pulps by the EG treatment were also discussed by studying the change in fibers characteristics and fiber bonding.

Unlabelled: Bacillus subtilis subsp. niger spore and Staphylococcus albus are typical biological indicators for the inactivation of airborne pathogens. The present study characterized and compared the behaviors of B. subtilis subsp. niger spores and S. albus in regard to inactivation by chlorine dioxide (ClO2) gas under different gas concentrations and relative humidity (RH) conditions. The inactivation kinetics under different ClO2 gas concentrations (1 to 5 mg/liter) were determined by first-order and Weibull models. A new model (the Weibull-H model) was established to reveal the inactivation tendency and kinetics for ClO2 gas under different RH conditions (30 to 90%). The results showed that both the gas concentration and RH were significantly (P < 0.05) and positively correlated with the inactivation of the two chosen indicators. There was a rapid improvement in the inactivation efficiency under high RH (>70%). Compared with the first-order model, the Weibull and Weibull-H models demonstrated a better fit for the experimental data, indicating nonlinear inactivation behaviors of the vegetative bacteria and spores following exposure to ClO2 gas. The times to achieve a six-log reduction of B. subtilis subsp. niger spore and S. albus were calculated based on the established models. Clarifying the kinetics of inactivation of B. subtilis subsp. niger spores and S. albus by ClO2 gas will allow the development of ClO2 gas treatments that provide an effective disinfection method., Importance: Chlorine dioxide (ClO2) gas is a novel and effective fumigation agent with strong oxidization ability and a broad biocidal spectrum. The antimicrobial efficacy of ClO2 gas has been evaluated in many previous studies. However, there are presently no published models that can be used to describe the kinetics of inactivation of airborne pathogens by ClO2 gas under different gas concentrations and RH conditions. The first-order and Weibull (Weibull-H) models established in this study can characterize and compare the behaviors of Bacillus subtilis subsp. niger spores and Staphylococcus albus in regard to inactivation by ClO2 gas, determine the kinetics of inactivation of two chosen strains under different conditions of gas concentration and RH, and provide the calculated time to achieve a six-log reduction. These results will be useful to determine effective conditions for ClO2 gas to inactivate airborne pathogens in contaminated air and other environments and thus prevent outbreaks of airborne illness., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Environmental contamination from petroleum refinery operations has increased due to the rapid population growth and modernization of society, necessitating urgent repair. Microbial remediation of petroleum wastewater by prominent bacterial cultures holds promise in circumventing the issue of petroleum-related pollution. Herein, the bacterial culture was isolated from petroleum-contaminated sludge samples for the valorization of polyaromatic hydrocarbons and biodegradation of petroleum wastewater samples. The bacterial strain was screened and identified as Bacillus subtilis IH-1. After six days of incubation, the bacteria had degraded 25.9% of phenanthrene and 20.3% of naphthalene. The treatment of wastewater samples was assessed using physico-chemical and Fourier-transform infrared spectroscopy analysis, which revealed that the level of pollutants was elevated and above the allowed limits. Following bacterial degradation, the reduction in pollution parameters viz. EC (82.7%), BOD (87.0%), COD (80.0%), total phenols (96.3%), oil and grease (79.7%), TKN (68.8%), TOC (96.3%) and TPH (52.4%) were observed. The reduction in pH and heavy metals were also observed after bacterial treatment. V. mungo was used in the phytotoxicity test, which revealed at 50% wastewater concentration the reduction in biomass (30.3%), root length (87.7%), shoot length (93.9%), and seed germination (30.0%) was observed in comparison to control. When A. cepa root tips immersed in varying concentrations of wastewater samples, the mitotic index significantly decreased, suggesting the induction of cytotoxicity. However, following the bacterial treatment, there was a noticeable decrease in phytotoxicity and cytotoxicity. The bacterial culture produces lignin peroxidase enzyme and has the potential to degrade the toxic pollutants of petroleum wastewater. Therefore the bacterium may be immobilised or directly used at reactor scale or pilot scale study to benefit the industry and environmental safety., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The improper treatment of crop straw not only leads to resource wastage but also adversely impacts the ecological environment. However, the application of microorganisms can accelerate the decomposition of crop straw and improve its utilization. In this study, cellulose-degrading microbial strains were isolated from naturally decayed corn straw and screened using Congo red staining, along with assessing variations in carboxymethyl cellulase (CMCase) activity, filter paper enzyme (FPase) activity and β-glucosidase (β-Gase) activity, as well as the degradation rate. The eight strains, namely Neurospora intermedia isolate 29 (A1), Streptomyces isolate FFJC33 (A2), Gibberella moniliformis isolate FKCB-009 (A3), Fusarium fujikuroi isolate EFS3(2) (A4), Fusarium Fujikuroi isolate FZ04 (A5), Lysine bacillus macroides strain LNHL43 (B1), Bacillus subtilis strain MPF30 (B2) and Paenibacilli lautus strain ALEB-P1 (C), were identified and selected for microbial strain consortium design based on their high activities of CMCase, FPase and β-Gase. The fungi, bacteria and actinomycete strains were combined without antagonistic effects on corn straw decomposition. The results showed the A2B2 combination had a significantly higher FPase at 55.44 U/mL and β-Gase at 25.73 U/mL than the other two strain combinations (p < 0.05). Additionally, the degradation rate of this combination was 40.33%, which was considerably higher than that of the other strains/consortia. The strain combination A4B2C also had superior enzyme activity, including CMCase with a value of 35.03 U/mL, FPase with a value of 63.59 U/mL and β-Gase with a value of 26.15 U/mL, which were significantly different to those of the other three strain combinations (p < 0.05). Furthermore, seven single microbial strains with high cellulase activities were selected to construct various microbial consortiums for in situ composting in order to evaluate their potential. Taken as a whole, the results of composting, including temperature, moisture content, pH, E4/E6 value and seed germination index, indicated that the microbial strain consortium consisting of Neurospora intermediate isolate 29, Fusarium fujikuroi isolate EFS3(2), Fusarium fujikuroi isolate FZ04, Lysinibacillus macrolides, Lysinibacillus sphaericus, Bacillus subtilis and Paenibacillus lautus was advantageous for corn straw decomposition and yielded high-quality compost. The screened flora was able to effectively degrade corn straw. This study provides a novel solution for the construction of a microbial consortium for the composting of corn straw. [ABSTRACT FROM AUTHOR]

Rapid and accurate bioburden detection has become increasingly necessary for food, health, pharmaceutical and environmental applications. To detect bioburden accurately, and in a highly sensitive manner, we have fabricated a novel microfluidic device with an integrated filter to trap the cells. Bioburden is detected on the filter paper in situ using the redox reaction of fluorescent label resorufin and a portable multichannel fluorometer is used for fluorescence measurement. The microfluidic device was fabricated in a facile, low-cost, and rapid way with microwave-induced thermally assisted bonding. To characterize the bonding quality of the microfluidic cassettes, different tests were performed, and the filter paper material and size were optimized. Primary Bacillus subtilis culture bacterial samples were filtered through the device to validate and investigate the performance parameters. Our results show that a limit of detection (LOD) of 0.037 CFU/mL can be achieved through this microfluidic device whereas the LOD in a normal microfluidic cassette in the fluorometer and the golden standard spectrophotometer are 0.378 and 0.128 CFU/mL respectively. The results depict that three to ten times LOD improvement is possible through this microfluidic cassette and more sensitive detection is possible depending on the volume filtered within a rapid 3 min. This novel microfluidic device along with the fluorometer can be used as a rapid portable tool for highly sensitive, accurate and high-throughput bacterial detection for different applications. [ABSTRACT FROM AUTHOR]

The potential of alkaline cellulo-xylanolytic enzymes from non-pathogenic Bacillus subtilis strain was tested for deinking of photocopier waste paper. Cellulase and xylanase play a crucial role in deinking of different types of waste paper. Partial purification of cellulo-xylanolytic enzymes was carried out using ultrafiltration followed by ammonium sulfate precipitation. The ultrafiltered enzyme was used for deinking the photocopier waste paper along with chemical deinking. An enzyme dose of 0.6 IU/g and reaction time of 60 min for ultrafiltered cellulo-xylanolytic enzyme significantly increased deinking efficiency, tear index (9.52%) and folding endurance (5±2%) as compared to chemical deinking. There was improvement in strength properties such as tear index and double-fold along with freeness of pulp (18%). There was slight decrease in tensile index (0.6%) and burst index (16%) while ISO brightness remained unaffected. Enzymatic deinking (74.3%) by ultrafiltered cellulo-xylanolytic from Bacillus subtilis was found significant over conventional chemical deinking. [ABSTRACT FROM AUTHOR]

Climate change is one of the most urgent environmental challenges that humanity faces. In addition to the reduction of greenhouse gas emissions, safe and robust carbon dioxide removal (CDR) technologies that capture atmospheric CO 2 and ensure long-term sequestration are required. Among CDR technologies, enhanced silicate weathering (ESW) has been suggested as a promising option. While ESW has been demonstrated to depend strongly on pH, water, and temperature, recent studies suggest that biota may accelerate mineral weathering rates. Bacillus subtilis is a plant growth-promoting rhizobacterium that can facilitate weathering to obtain mineral nutrients. It is a promising agricultural biofertilizer, as it helps plants acquire nutrients and protects them from environmental stresses. Given that croplands are optimal implementation fields for ESW, any synergy between ESW and B. subtilis can hold great potential for further practice. B. subtilis was reported to enhance weathering under laboratory conditions, but there is a lack of data for soil applications. In a soil-mesocosm experiment, we examined the effect of B. subtilis on basalt weathering. B. subtilis-basalt interaction stimulated basalt weathering and increased soil extractable Fe. The combined application displayed higher CDR potential compared to basalt-only application (3.7 vs. 2.3 tons CO 2 ha -1 ) taking solid and liquid cation pools into account. However, the cumulative CO 2 efflux decreased by approximately 2 tons CO 2 ha -1 with basalt-only treatment, while the combined application did not affect the CO 2 efflux. We found limited mobilization of cations to the liquid phase as most were retained in the soil. Additionally, we found substantial mobilization of basalt-originated Mg, Fe, and Al to oxide- and organic-bound soil fractions. We, therefore, conclude that basalt addition showed relatively low inorganic CDR potential but a high capacity for SOM stabilization. The outcomes indicated the importance of weathering rate-GHG emission integration and the high potential of SOM stabilization in ESW studies., (© 2024 John Wiley & Sons Ltd.)

The aim of the present study is to evaluate the probiotic attributes of Bacillus subtilis AMS6 isolated from fermented soybean (Churpi). This isolate exhibited tolerance to low pH (pH 2.0) and bile salt (0.3%), capability to autoaggregate and coaggregate. AMS6 also showed highest antibacterial activity against the pathogenic indicator strain Salmonella enterica typhimurium (MTCC 1252) and susceptibility towards different antibiotics tested. The isolate was effective in inhibiting the adherence of food borne pathogens to Caco-2 epithelial cell lines, and was also found to be non-hemolytic which further strengthen the candidature of the isolate as a potential probiotic. Further studies revealed B. subtilis AMS6 showed cellulolytic activity (0.54±0.05 filter paper units mL(-1)) at 37°C. The isolate was found to hydrolyze carboxymethyl cellulose, filter paper and maize (Zea mays) straw. The maize straw digestion was confirmed by scanning electron microscopy studies. The isolate was able to degrade filter paper within 96h of incubation. A full length cellulase gene of AMS6 was amplified using degenerate primers consisting of 1499 nucleotides. The ORF encoded for a protein of 499 amino acids residues with a predicted molecular mass of 55.04kDa. The amino acids sequence consisted of a glycosyl hydrolase family 5 domain at N-terminal; Glycosyl hydrolase catalytic core and a CBM-3 cellulose binding domain at its C terminal. The study suggests potential probiotic B. subtilis AMS6 as a promising candidate envisaging its application as an animal feed additive for enhanced fiber digestion and gut health of animal., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

An abundance of refractory cellulose is the key limiting factor restricting the resource utilization efficiency of silkworm (Bombyx mori) excrement via composting. Screening for cellulose-degrading bacteria is likely to provide high-quality strains for the safe and rapid decomposition of silkworm excrement. In this study, bacteria capable of degrading cellulose with a high efficiency were isolated from silkworm excrement and the conditions for cellulase production were optimized. The strains were preliminarily screened via sodium carboxymethyl cellulose culture and staining with Congo red, rescreened via a filter paper enzyme activity test, and identified via morphological observation, physiological and biochemical tests, and phylogenetic analysis of the 16S rDNA sequence. Enzyme activity assay was performed using the 3,5-dinitrosalicylic acid method. DC-11, a highly cellulolytic strain, was identified as Bacillus subtilis. The optimum temperature and pH of this strain were 55 °C and 6, respectively, and the filter paper enzyme activity (FPase), endoglucanase activity (CMCase), and exoglucanase activity (CXase) reached 15.40 U/mL, 11.91 U/mL, and 20.61 U/mL. In addition, the cellulose degradation rate of the treatment group treated with DC-11 was 39.57% in the bioaugmentation test, which was significantly higher than that of the control group without DC-11 (10.01%). Strain DC-11 was shown to be an acid-resistant and heat-resistant cellulose-degrading strain, with high cellulase activity. This strain can exert a bioaugmentation effect on cellulose degradation and has the potential for use in preparing microbial inocula that can be applied for the safe and rapid composting of silkworm excrement. [ABSTRACT FROM AUTHOR]

Exploring an appropriate immobilization approach to enhance catalytic activity and reusability of cellulase is of great importance to reduce the price of enzymes and promote the industrialization of cellulose-derived biochemicals. In this study, Fe3O4 magnetic nanoparticles (MNPs) were functionalized with meso-2,3-dimercaptosuccinic acid to introduce carboxyl groups on the surface (DMNPs). Then, melamine–glutaraldehyde dendrimer-like polymers were grafted on DMNPs to increase protein binding sites for the immobilization of processive endoglucanase EG5C-1. Moreover, this dendrimer-like structure was beneficial to protect the conformation of EG5C-1 and facilitate the interaction between substrate and active center. The loading capacity of the functionalized copolymers (MG-DMNPs) for EG5C-1 was about 195 mg/g, where more than 90% of the activity was recovered. Immobilized EG5C-1 exhibited improved thermal stability and increased tolerability over a broad pH range compared with the free one. Additionally, MG-DMNP/EG5C-1 biocomposite maintained approximately 80% of its initial hydrolysis productivity after five cycles of usage using filter paper as the substrate. Our results provided a promising approach for the functionalization of MNPs, enabling the immobilization of cellulases with a high loading capacity and excellent activity recovery. [ABSTRACT FROM AUTHOR]

The aim of this work was to demonstrate the process of manufacturing and applying functionalising additives using a pilot research installation. As part of the optimisation evaluation, hygienic papers functionalised with biopolymer-biocide compositions based on chitosan, starch and Gemini surfactants were prepared. The microbiological properties and susceptibility to biodegradation of prototype hygienic papers (prepared in EPICOM) were assessed In particular, the minimum amount of biopolymer and bioactive agent were determined to ensure the finished product exerted antibacterial activity against Staphylococcus aureus, Bacillus subtilis and Escherichia coli and was more than 90% biodegradable in a compost environment. Under optimal conditions, the hygienic paper exerted excellent antibacterial activity against S. aureus and E. coli as well as good activity against B. subtilis. Moreover, the hygienic paper showed more than 90% biodegradability in compost conditions within 8 weeks. [ABSTRACT FROM AUTHOR]

Bacillus subtilis is regarded as a promising microbial expression system in bioengineering due to its high stress resistance, nontoxic, low codon preference and grow fast. The strain has a relatively efficient expression system, as it has at least three protein secretion pathways and abundant molecular chaperones, which guarantee its expression ability and compatibility. Currently, many proteins are expressed in Bacillus subtilis, and their application prospects are broad. Although Bacillus subtilis has great advantages compared with other prokaryotes related to protein expression and secretion, it still faces deficiencies, such as low wild-type expression, low product activity, and easy gene loss, which limit its large-scale application. Over the years, many researchers have achieved abundant results in the modification of Bacillus subtilis expression systems, especially the optimization of promoters, expression vectors, signal peptides, transport pathways and molecular chaperones. An optimal vector with a suitable promoter strength and other regulatory elements could increase protein synthesis and secretion, increasing industrial profits. This review highlights the research status of optimization strategies related to the expression system of Bacillus subtilis. Moreover, research progress on its application as a food-grade expression system is also presented, along with some future modification and application directions., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aims to investigate the antimicrobial activity of paper-based packaging materials including coriander (Coriandrum sativum L.) essential oil. Three types of packaging papers (bleached, unbleached, and recycled) are treated with an essential oil and then stored for 2 h, 24 h, and 5 days. The antibacterial activity of coriander oil is tested against Gram-positive bacteria (Staphylococcus aureus ATCC 6538 and Bacillus subtilis ATCC 6633); Gram-negative bacteria (Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, and Salmonella abony NTCC 6017); an yeast (Candida albicans ATCC 10231), and a fungal strain (Aspergillus brasiliensis ATCC 16404). A high fungicidal potential is observed in case of all packaging materials. The antimicrobial action of the bleached paper coated with coriander oil against Gram-positive bacteria during the five-day storage period decreases slightly from 100 % - 94 % to 79 % - 81 %. [ABSTRACT FROM AUTHOR]

Biofilm formation is a process in which microbial cells aggregate to form collectives that are embedded in a self-produced extracellular matrix. Bacillus subtilis is a Gram-positive bacterium that is used to dissect the mechanisms controlling matrix production and the subsequent transition from a motile planktonic cell state to a sessile biofilm state. The collective nature of life in a biofilm allows emergent properties to manifest, and B. subtilis biofilms are linked with novel industrial uses as well as probiotic and biocontrol processes. In this Review, we outline the molecular details of the biofilm matrix and the regulatory pathways and external factors that control its production. We explore the beneficial outcomes associated with biofilms. Finally, we highlight major advances in our understanding of concepts of microbial evolution and community behaviour that have resulted from studies of the innate heterogeneity of biofilms., (© 2021. Springer Nature Limited.)

Colorimetric, fluorescence, and paper-based method were developed to measure the Hg2+ level in water using iturin A, a lipopeptide produced by Bacillus subtilis. Firstly, iturin was used to synthesize highly stable and uniformly sized silver nanoparticles (AgNPs). Secondly, the iturin-AgNPs were found to be highly selective and sensitive to Hg2+. The absorbance of the reaction system showed a good linear correlation with the Hg2+ concentration from 0.5 to 5 mg/L at 450 nm in the UV-Vis spectroscopy detection with the limit of detection (LOD) of 0.5 mg/L. When the reaction system was detected by fluorescence measurement, a good linear relationship was found between the fluorescence intensity and Hg2+ concentration from 0.05 to 0.5 mg/ at 415 nm with the LOD of 0.05 mg/L. Lastly, a paper-based detection method was developed. The developed method was successfully used to detect Hg2+ in contaminated polluted waters and showed acceptable results in terms of sensitivity, selectivity and stability. The paper-based method could distinguish Hg2+ at levels higher than 0.05 mg/L, thereby meeting the guidelines of the effluent quality standard for industries (0.05 mg/L). In summary, this method can be used daily by various industries to monitor the Hg2+ level in effluent water. [ABSTRACT FROM AUTHOR]

The worldwide availability of lignocellulosic wastes represents a serious environmental challenge with potential opportunities. Xylanases are crucial in lignocellulosic bio-hydrolysis, but the low enzyme productivity and stability are still challenges. In the current study, Bacillus subtilis (coded ARSE2) revealed potent xylanase activity among other local isolates. The enzyme production optimization revealed that maximum enzyme production (490.58 U/mL) was achieved with 1% xylan, 1.4% peptone, and 5% NaCl at 30 °C and pH 9. Furthermore, several lignocellulosic wastes were exploited for sustainable xylanase production, where sugarcane bagasse (16%) under solid-state fermentation and woody sawdust (2%) under submerged fermentation supported the maximum enzyme titer of about 472.03 and 485.7 U/mL, respectively. The partially purified enzyme revealed two protein bands at 42 and 30 kDa. The partially purified enzyme revealed remarkable enzyme activity and stability at 50-60 °C and pH 8-9. The enzyme also revealed significant stability toward tween-80, urea, DTT, and EDTA with V max and K m values of 1481.5 U/mL and 0.187 mM, respectively. Additionally, the purified xylanase was applied for xylooligosaccharides production, which revealed significant antimicrobial activity toward Staphylococcus aureus with lower activity against Escherichia coli. Hence, the locally isolated Bacillus subtilis ARSE2 could fulfill the xylanase production requirements in terms of economic production at a high titer with promising enzyme characteristics. Additionally, the resultant xylooligosaccharides revealed a promising antimicrobial potential, which paves the way for other medical applications., (© 2023. The Author(s).)

In this study, the coupling of magnetic enrichment of bacteria from real samples with rapid surface enhanced Raman spectroscopy (SERS) detection was reported. The selective isolation and enrichment for the model bacteria Escherichia coli (E. coli) was performed using E. coli (primary) antibody bound-magnetic gold (Fe 3 O 4 @Au) nanoparticles. Following isolation and enrichment, the rennet enzyme was used to cleave of casein modified Fe 3 O 4 /Au-PEI nanoparticles from primary antibody-bound bacteria to prevent the nanoparticle aggregation and provide the movement of bacteria on nitrocellulose membrane. In the first part of the study, optimization studies were carried out namely; the amounts of gold nanoparticles (AuNPs), polyethyleneimine coated magnetic gold (Fe 3 O 4 /Au-PEI) nanoparticles, casein and rennet enzyme. The SERS signals of DTNB (5,5′-Dithiobis(2-nitrobenzoic acid)) molecule were collected on the test line and a calibration curve was plotted by using signal intensities. The correlation between the concentration of E. coli and SERS signal was found to be linear within the range of 101–107 cfu/mL (R2 = 0.984, LOD = 0.52 cfu/mL and LOQ = 1.57 cfu/mL). The selectivity of the paper-based lateral flow immunoassay (LFIA) was examined with Bacillus subtilis (B. subtilis), Micrococcus luteus (M. luteus), Salmonella enteritidis (S. enteritidis) which did not produce any significant response compared with E. coli measurement. Finally, the developed paper-based LFIA was tested with urine and milk samples. The obtained SERS results were compared with a plate counting method results which were in a good accordance. The developed method was found as rapid and sensitive to E. coli with a total analysis time of less than 60 min. Image 1 • Paper-based strips were developed using the magnetic enrichment. • LFIA strips were performed using rennet enzyme isolation of E. coli from buffer and real media. • The selectivity tests of LFIA were obtained with milk and urine media. • The specificity tests of LFIA were evaluated with B. subtilis , M. luteus , S. enteritidis. [ABSTRACT FROM AUTHOR]

Lateral flow immunoassays (LFIs) can be used to detect intact bacteria or spores; when gold nanoparticles (AuNPs) are used as the signal reporters, the detection limits are very low. Spore-based surface display has been widely studied for enzyme immobilization and live-nontoxic oral vaccines. In this study, recombinant spores were used to improve the sensitivity of a LFI. We developed a test kit that combines streptavidin-displayed spores with a LFI assay for rapid protein detection. The recombinant spores served as a signal amplifier and AuNPs were used as the signal reporters. For detection of β-galactosidase, which was used as the model protein, the detection limit was about 10 -15  mol, while that of the conventional LFI is about 10 -12  mol. In both methods, nanogold was used as the colorimetric signal and could be observed with the naked eye. This method improved LFI sensitivity without sacrificing its advantages. Furthermore, enhanced green fluorescent protein (eGFP) was also displayed on the surface of the streptavidin-displayed spores. Without AuNPs, the fluorescent recombinant spores acted as the signal, which could be detected by a fluorescence detector, such as a fluorescence microscope. The detection limit was 10 -16  mol under fluorescence microscopy whose magnification was 25-fold. Therefore, in conclusion, in this proof of concept study, the detection limits of both proposed methods were far superior to those of traditional LFI assay.

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In either a living system or a non-living system, the interaction among its constituent cells or particles is a fundamental aspect at all scales. For example, during the Bacillus subtilis biofilm formation, cells differentiate into multiple phenotypes to adapt to the environments; few hours after the initial inoculation, we find the phenotype of matrix-producing cells form "chain" structure surrounding the phenotype of the "running" motile cells. We use "chain" to characterize the structure of matrix-producing cells, and "running" to characterize the proliferation and growth of motile cells. Due to a large number of cells in the biofilm, it is impossible to construct a traditional kinetic model to describe the causal link between the single-cell movement and the colony behavior. Here, we obtain cell state information and cell group shape information through experiments; after the image analysis, we get the key interaction rules between cells, and then, we simulate the comparable movement of two cell types and the resulting colony geometry using the multi-agent model. Our work makes a better understanding of the relationship between the macroscopic shape of colonies and microscopic mechanical interactions among cells in the early stage of biofilm growth., (© 2021. European Biophysical Societies' Association.)

The effectiveness of inexpensive agro-industrial wastes such as sugarcane molasses, paper mill effluent, and dairy effluent for the production of polyhydroxyalkanoates (PHAs) using Bacillus subtilis RS1 was assessed in this study. Laboratory-scale fermentations were carried out with both raw and pre-treated agro-industrial effluents, and maximum PHAs yield was observed in pre-treated effluents. Experimental variables such as pH, inoculum dose, and incubation time were further optimized to maximize PHAs yield, and the optimal conditions were identified as incubation time 48 h; pH 7; and inoculum dose 10% v/v. PHAs yield in pre-treated sugarcane molasses increased to 70.5% under optimal conditions. Fourier transform infrared spectroscopy and gas chromatography–mass spectrometric analysis revealed that the extracted polymer was composed of penta- and hexadecanoic acid methyl esters, a copolymer of PHAs. Results indicated that the pre-treated sugarcane molasses could be used as an inexpensive substrate for the production of PHAs. [ABSTRACT FROM AUTHOR]

Bioremediation is widely used to remove water pollution as environmentally friendly smart solutions. In this study, Bacillus isolates were investigated in terms of the effectiveness of single and multiple cultures in eliminating aquatic pollution related to aquaculture activities. In the established experimental setups, the environments where Bacillus isolates were inoculated with single and multiple cultures at 1 × 10 7  CFU/mL were evaluated comparatively with control groups without these isolates, and total aerobic mesophilic bacterial counts were performed in the petri dish by inoculation method. At the end of the 6 days of the experiment, in the environment in which single and multiple cultures of Bacillus isolates were presented with 17-20 ± 0.05 °C temperature and 5.1-8.1 pH 2-4.6 mg/l dissolved oxygen values (O 2 ), 2% increase in total phosphorus (TP) value was observed. On the other hand, 4% removal of Ammonia-nitrogen (NH 3 -N), 80% removal of Nitrite-nitrogen (NO 2 -N), and 100% removal of Nitrate-nitrogen (NO 3 -N) were observed. In the changes in heavy metal concentrations, the removal of Ni, Cr, Se, Al, Cd, Mn, Fe, and B was observed from highest to lowest as 57%, 50%, 50%, 43%, 40%, 23%, 5%, and 2%, respectively. It also has been seen that B. thuringiensis isolate was observed to be more effective than B. subtilis in metal removal.

Bacillus subtilis subsp. niger spore and Staphylococcus albus are typical biological indicators for the inactivation of airborne pathogens. The present study characterized and compared the behaviors of B. subtilis subsp. niger spores and S. albus in regard to inactivation by chlorine dioxide (ClO2) gas under different gas concentrations and relative humidity (RH) conditions. The inactivation kinetics under different ClO2 gas concentrations (1 to 5 mg/liter) were determined by first-order and Weibull models. A new model (the Weibull-H model) was established to reveal the inactivation tendency and kinetics for ClO2 gas under different RH conditions (30 to 90%). The results showed that both the gas concentration and RH were significantly (P<0.05) and positively correlated with the inactivation of the two chosen indicators. There was a rapid improvement in the inactivation efficiency under high RH (>70%). Compared with the first-order model, the Weibull and Weibull-H models demonstrated a better fit for the experimental data, indicating nonlinear inactivation behaviors of the vegetative bacteria and spores following exposure to ClO2 gas. The times to achieve a six-log reduction of B. subtilis subsp. niger spore and S. albus were calculated based on the established models. Clarifying the kinetics of inactivation of B. subtilis subsp. niger spores and S. albus by ClO2 gas will allow the development of ClO2 gas treatments that provide an effective disinfection method. [ABSTRACT FROM AUTHOR]