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To breakthrough the delignification saturation point (DSP) of alkaline hydrogen peroxide (AHP) pretreatment, a biphasic AHP/pentanol (AHPP) pretreatment was proposed in this work. The temperature and H 2 O 2 concentration were evaluated. Under the optimal conditions (110 °C, 2 h, 4 % H 2 O 2 ), 70.73 % of lignin was removed, which was increased by 11.65 % than the traditional AHP pretreatment, indicating successful overcoming of the DSP by adding pentanol. 85.74 % and 88.62 % of glucan and xylan digestibility were achieved, respectively, which increased by 7.41 % and 5.87 % as compared to AHP pretreatment. Furthermore, the lignin extracted from the organic phase accounted for 38.51 % of the delignification, and it had a low molecular weight, effectively preserving the β-O-4 bonds. Finally, satisfied pentanol recovery (77.91 %) and delignification (57.19 %) along with excellent glucan (76.11 %) and xylan (77.52 %) digestibility were reached after fourth recycling of AHPP pretreatment. Therefore, AHPP pretreatment was a promising method for biomass valorization within biorefinery concept., 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.)

Esculin is a polyphenol with multiple bioactivities and poor lipophilicity. Therefore, a whole-cell catalytic strategy for esculin acylation was developed to improve its lipophilicity. A total of 12 strains were tested, among which Pseudomonas stutzeri exhibited the highest catalytic activity and mono-acylated regioselectivity. The conversion reached the highest level of 92.7 % at 24 h under the optimal conditions, when vinyl acetate was used as an acyl donor. The catalytic ability of P. stutzeri remained above 60 % after three cycles. Subsequently, five esculin esters with different lengths of fatty chains were synthesized and structurally identified. Of them, esculin-6'-O-octanoate, esculin-6'-O-laurate, and esculin-6'-O-myristate exhibited cytotoxicity on LO2 cells by inducing apoptosis and necrosis. The cytotoxicity of these three esters may attribute to their membrane-disrupting properties. This study provides a novel whole-cell biocatalytic strategy for the acylation of esculin and insight for application of esculin esters as a food additive or drug., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Comparative studies of the effects of two commercial enzyme formulations on fiber refining were conducted. Extensive basic characterisation of the enzymes involved, assessment of their hydrolytic activities on different model substrates as well as on different pulps (softwood sulfate, softwood sulfite, hardwood sulfate) were evaluated. Both enzyme formulations showed endoglucanase as well as some xylanase and β-glucosidase activity. In addition, Enzyme A reached a CMC end viscosity of 19.5 mPa compared to 11.1 mPa for Enzyme B. Reducing sugar release almost doubled from 695 μmol mL -1 for hardwood sulfate pulp to 1300 μmol mL -1 for softwood sulfite pulp with Enzyme B under the same conditions. Enzyme A increased the degree of refining even under non-ideal conditions from 23 °SR to up to 50 °SR. Further characterization of hand sheets, made from enzyme pre-treated and refined cellulose fibers with Enzyme A and B, showed that Enzyme A had the best effects leading to hand sheets with increased tensile strength and low air permeability. In summary, the increase in the degree of refining seen for Enzyme A correlated to higher xylanase and β-glucosidase activity and lower endoglucanase activity., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Catalytic transformation of biomass-derived furans into value-added chemicals and biofuels has received considerable interest recently. In this work, aldehyde dehydrogenases (ALDHs) were identified from Comamonas testosteroni SC1588 for the oxidation of bio-based furans into furan carboxylic acids. Of the whole-cell biocatalysts constructed, Escherichia coli expressing a vanillin dehydrogenase (E. coli_CtVDH1) proved to be the best for the oxidation of 5-hydroxymethylfurfural (HMF). 5-Hydroxymethyl-2-furancarboxylic acid (HMFCA) was obtained in the yield of approximately 92 % within 12 h using this recombinant strain when the HMF concentration was up to 200 mM. In a fed-batch process, 292 mM of HMFCA was produced within 20.5 h, thereby providing a productivity as high as 2.0 g/L h. Other furan carboxylic acids were synthesized in the yields of 83-95%. Besides, the partially purified HMF was smoothly converted into HMFCA by this recombinant strain, with a 90% yield., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Highly efficient asymmetric reduction of 2-octanone to (R)-2-octanol catalyzed by immobilized Acetobacter sp. CCTCC M209061 cells was achieved in a biphasic system. Bioreduction conducted in aqueous single phase buffer was limited due to poor solubility and toxicity towards cells cause by product accumulation. Introduction of [C 4 MIM]·Ac accelerated the biotransformation process, giving 99% yield, >99% product e.e. and 1.42-fold higher initial reaction rate in conversion of 10 mM 2-octanone as substrate, compared with 99% yield, 97.3% product e.e. and 1.57 μmol min -1 of initial reaction rate in a aqueous single phase buffer system containing 6 mM 2-octanone as the optimal substrate concentration. Moreover, in the [C 4 MIM]·Ac-containing buffer/n-tetradecane biphasic system, the optimal substrate concentration was enhanced by 83 times (500 mM) in comparison with that in aqueous single phase buffer, resulting in 53.4% yield (267 mM), 99% product e.e. with 8.9 mM g -1  h -1 space time yield., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The stabilization effects on a novel commercial β-glucosidase preparation from Aspergillus fumigatus during saccharification of ethanol-water pretreated wheat straw were analysed in comparison to this enzyme stability during cellobiose hydrolysis. For this purpose, the kinetics of β-glucosidase residual activity during cellobiose hydrolysis from 40 till 70 °C were studied, resulting in the fitting of a first-order partial deactivation model. Furthermore, a subsequent fitting of a kinetic model including this first-order deactivation equation and a Michaelis-Menten equation with double competitive inhibition by glucose and uncompetitive inhibition by cellobiose to released glucose was successful. Finally, global enzyme deactivation and prospective deactivation of enzyme remaining in the liquid phase were evaluated during wheat straw hydrolysis at 50 °C as a relevant saccharification process. Results suggest that the presence of a solid substrate dramatically reduces the global deactivation rate of the enzyme and, in addition, there is no loss the stability of the enzyme in the liquid phase along the saccharification process, even for 72 h., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The scientific community, experts in technology and marketing have been seeking cost-competitive and green solvents with good dissolving capacity for the valorisation of biomass and biowaste. Along with traditional solvents and techniques, deep eutectic solvents (DESs) and their bio-analogues, natural deep eutectic solvents (NADESs) are currently emerging as a new class of promising liquid media. In this review, a comprehensive summary of recent contribution of DESs to the processing and valorisation of various kinds of plant and animal based biomass and biowaste is provided. In the field of food industry based and agro-forest waste valorisation, through treatment of the waste, by-products, and natural materials by DESs, several types of compounds, such as flavonoids and other plant phenolics, phenolic acids, stilbenes, tannins, lignans, and lignin were obtained. Extraction of algae by DESs led to isolation mainly of proteins, carbohydrates, lipids and nucleic acids. Vegetable oils, spent oils, residues and by-products of their processing are a rich source of phenolic compounds such as phenolic acids and phenolic alcohols, secoiridoid derivatives (aglycone and ligstroside), lignans (pino and acetoxypinoresinol), flavones (luteolin and apigein), tocopherols and tocotrienols. Dietary fibre serves as a source of lignin, pectic substances, gums, resistant starch, inulin, as well as non-carbohydrate components, e.g., polyphenols, waxes, saponins, cutin, phytates, resistant proteins. Valorisation of wastes originated from animal processing by DESs means obtaining high-value chemicals including amino acids, proteins, bioactive peptides, collagen peptides, albumin etc. Through the valorisation of the mentioned waste types value-added products with potential applications in the pharmaceutical, food and cosmetic industries are produced. The paper gathered data on the used DESs, treated substances and obtained products, together with treatment conditions and the products yields. The evaluation of the state-of-the-art in the field of biowaste valorisation using DESs and NADESs led to conclusions and indication of future prospects and predicted development in this field., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The current study focuses on the analysis of in vitro biological activity of extract from bark of Norway spruce (Picea Abies), which can find potential application in food and cosmetic industry and pharmacology. Milled bark was subjected to Soxhlet extraction and supercritical fluid extraction to obtain two ethanol extracts. These extracts were further used to obtain their pre-extracts to n-hexane. It was investigated whether beta-sitosterol exhibits bacteriostatic activity necessary to observe antimicrobial and antifungal activity of methyl dehydroabiatate. This synergic effect and bacteriostatic activity of beta-sitosterol have not been previously reported. The greatest inhibition zone of n-hexane pre-extracts was confirmed in bacterium Pseudomonas aeruginosa (0,9 - 1,5 cm) and yeast Alternaria alternata (0,7 - 1,6 cm). It is novel, the antioxidant, antimicrobial and antifungal activity of spruce bark extracts assessed in terms of food and cosmetic fortification., (Copyright © 2018 Elsevier B.V. All rights reserved.)

An effective method to functionalize chitosan with 4-hexyloxyphenol (HP) under homogeneous reaction conditions was developed using laccase as the catalyst. The resulting copolymer was characterized for chemical structure, grafted-HP content, surface morphology, thermal stability, antioxidant capacity, hydrophobic properties and tensile strength. Solid-state 13 C NMR spectrum confirmed the incorporation of HP onto chitosan. X-ray diffraction (XRD) showed a decrease in the degree of crystallinity for laccase/HP treated chitosan compared to pure chitosan. The grafted-HP content in laccase/HP-treated chitosan first increased and then declined with increase of the initial HP/chitosan ratio. A heterogeneous surface with spherical particles on the laccase/HP treated chitosan was observed by environmental scanning electron microscopy (ESEM) and scanning probe microscopy (SPM). The laccase/HP treatment of chitosan improved the thermal stability of copolymer. More significantly, the HP functionalized chitosan showed greatly improved ABTS + and DPPH radicals scavenging capacity, compared with pure chitosan. The hydrophobicity property of the HP functionalized chitosan also significantly increased although its tensile strength decreased. This new type of composite with double functionalities (i.e., antioxidant and hydrophobic) could potentially be used as food packaging materials or coating agents., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Semi-synthetic β-lactam antibiotics (SSBAs) are one of the most important antibiotic families in the world market. Their enzymatic synthesis can be catalyzed by penicillin G acylases (PGAs). In this study, to improve enzymatic synthesis of ampicillin, site-saturating mutagenesis was performed on three conserved amino acid residues: βF24, αR146, and αF147 of thermo-stable penicillin G acylase from Alcaligenes faecalis (Af PGA). Four mutants βF24G, βF24A, βF24S, and βF24P were recovered by screening the mutant bank. Kinetic analysis of them showed up to 800-fold increased kcat/Km value for activated acyl donor D-phenylglycine methyl ester (D-PGME). When βF24G was used for ampicillin synthesis under kinetic control at industrially relevant conditions, 95% of nucleophile 6-aminopenicillanic acid (6-APA) was converted to ampicillin in aqueous medium at room temperature while 12% process time is needed to reach maximum product accumulation at 25% enzyme concentration compared with the wild-type Af PGA. Consequently, process productivity of enzymatic synthesis of ampicillin catalyzed by Af PGA was improved by more than 130 times, which indicated an enzyme viable for efficient SSBAs synthesis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Enzymatic regioselective acylation of pyrimidine nucleosides was mediated by immobilized lipase from Penicillium expansum in 2-methyltetrahydrofuran (MeTHF), a bio-solvent derived from biomass. Despite of the moderate dissolution ability of MeTHF toward nucleosides, the initial enzymatic reaction rate was much higher in this eco-friendly solvent than in other commonly used organic solvents. This could be explained by the lower apparent activation energy of the enzymatic reaction (24.5 vs. 43.3-57.1kJ/mol) and the higher catalytic efficiency of the enzyme (Vmax/Km, 5.8 vs. 1.1-2.9h(-1)) in MeTHF. The enzymatic acylation of a group of ribonucleosides afforded the desirable 5'-esters with the conversions of 96-99% and 5'-regioselectivities of 96 to >99%. In enzymatic acylation of 2'-deoxynucleosides, however, 5'-regioselectivities showed a clear dependence on the 5-substituents present in the base moiety although the substrate conversions reached >98% within 1-3h. In the cases of 2',3'-dideoxynucleoside analogs, the reaction rate decreased markedly due to the lack of 3'-hydroxyl., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A comparative study was made of Mung bean epoxide hydrolases-catalyzed asymmetric hydrolysis of styrene oxide to (R)-1-phenyl-1,2-ethanediol in an n-hexane/buffer biphasic system containing various hydrophilic ionic liquids (ILs). Compared to the n-hexane/buffer biphasic system alone, addition of a small amount of hydrophilic ILs reduced the amount of non-enzymatic hydrolysis, and improved the reaction rate by up to 22%. The ILs with cation containing an alkanol group, namely [C(2)OHMIM][BF(4)] and [C(2)OHMIM][TfO], and the choline amino acid ILs [Ch][Arg] and [Ch][Pro] were found to be the most suitable co-solvents for the reaction, owing to their good biocompatibility with the enzyme, which led to high initial rates (0.99-1.25 μmol/min) and high product e.e.s (95%). When substrate concentration was around 30 mM, where optimal performance was observed with the IL-containing systems, the product e.e. was improved from 90% without ILs to ≥95% in the presence of ILs., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Cationic polyacrylamides (c-PAMs) increase the binding of cellulase to cellulose and of α-amylase to cornstarch. The c-PAMs increase the rate of hydrolysis of both substrates by about the same amount, which suggests that the increased binding is responsible for the enhanced rates. These results support our proposed mechanism where the c-PAM neutralizes the charge of the substrate through a "patching" mechanism, which reduces the charge repulsion between fiber and enzyme., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The biocatalytic enantioselective reduction of 4'-methoxyacetophenone to (S)-1-(4-methoxyphenyl)ethanol was successfully conducted in a hydrophilic IL-containing co-solvent system using immobilized Rhodotorula sp. AS2.2241 cells. Of all the tested ILs, the best results were observed with the novel IL 1-(2'-hydroxy)ethyl-3-methylimidazolium nitrate (C(2)OHMIM.NO(3)), which showed a good biocompatibility with the cells and increased the cell membrane permeability moderately, thus improving the efficiency of the bioreduction. To better understand the bioreduction, several crucial influential variables were also examined. The optimal C(2)OHMIM.NO(3) content, buffer pH, reaction temperature and substrate concentration were 5.0% (v/v), 8.5, 25 degrees C and 12 mM, respectively. Under the optimized conditions, the initial reaction rate, the maximum yield and the product e.e. were 9.8 micromol/hg(cell), 98.3% and >99%, respectively, which are much better than the results previously reported. The established biocatalytic system has proven to be highly effective for the reduction of other aryl ketones. Also, the cells exhibited excellent operational stability in the presence of C(2)OHMIM.NO(3). Moreover, the ILs can accumulate within the cells, suggesting that ILs are likely to interact with the related enzymes within the cells.

Among the five vinyl esters bearing phenyl ring in acyl moiety, the lipase from Burkholderia cepacia was most specific toward vinyl 4-phenylbutyrate. Additionally, 3'-regioselectivity of the enzyme was enhanced with the increment of acyl chain length. The lipophilic 3'-arylaliphatic acid esters of floxuridine, its potential prodrugs, were synthesized with high conversions (>98%) and good to excellent regioselectivities (85-99%) via the enzymatic approach.

Recombinant Volvariella volvacea endoglucanase 1 (EG1) and its catalytic module (EG1-CM) were obtained by expression in Pichia pastoris, purified by two-step chromatography, and the catalytic activities and binding capacities were compared. EG1 and EG1-CM exhibited very similar specific activities towards the soluble substrates carboxymethyl cellulose, lichenan and mannan, and insoluble H(3)PO(4) acid-swollen cellulose, whereas the specific activities of EG1-CM towards the insoluble substrates alpha-cellulose, Avicel and filter paper were approximately 58, 43 and 38%, respectively compared to EG1. No increase in reducing sugar release was detected in the reaction mixture supernatants after 50h exposure of filter paper, Avicel or alpha-cellulose to EG1-CM, whereas increases in the total reducing sugar equivalents (i.e. reducing sugar released into solution together with new reducing ends generated in the cellulosic substrates) in reaction mixtures were observed after 1h. In reaction mixtures containing EG1, soluble reducing sugar equivalents were detected in supernatants after 3h incubation with the insoluble cellulosic substrates. EG1-CM did not adsorb to Avicel, and the binding capacities of EG1-CM towards filter paper and H(3)PO(4) acid-swollen cellulose were 27.9-33.3% and 29.6-60.6%, respectively of values obtained with EG1 within the range of total added protein. In enzymatic deinking experiments, the ink removal rate in EG1-CM-treated samples was only slightly higher (approximately 8%), than that of untreated controls, whereas that of the EG1-treated samples was 100% higher. Bio-stoning of denim with EG1-CM resulted in increases of 48% and 40% in weight loss and indigo dye removal, respectively compared with untreated controls. These increases were considerably lower than the corresponding values of 219% and 133% obtained when samples were treated with EG1.

Flax fibres (Linum usitatissimum L.) were subjected to chemical and enzymatic analysis in order to determine the compositional changes brought about by the retting process and also to determine the accessibility of the fibre polymers to enzymatic treatment. Chemical analysis involved subjecting both retted and non retted fibres to a series of sequential chemical extractions with 1% ammonium oxalate, 0.05 M KOH, 1 M KOH and 4 M KOH. Retting was shown to cause minimal weight loss from the fibres but caused significant changes to the pectic polymers present. Retted fibres were shown to have significantly lower amounts of rhamnogalacturonan as well as arabinan and xylan. In addition the average molecular mass of the pectic extracts was considerably lowered. Enzyme treatment of the 1 M KOH extracts with two different enzymes demonstrated that the non retted extract contained a relatively high molecular weight xylan not found in the retted extract. Treatment of the 1 M KOH extracts and the fibres with Endoglucanase V from Trichoderma viride demonstrated that while this enzyme solubilised cellulose as well as xylan and xyloglucan oligomers from the extract, it had limited access to these polymers on the fibre. MALDI-TOF MS analysis of the material solubilised from the extract suggested that the xylan was randomly substituted with 4-O-methyl glucuronic acid moieties. The xyloglucan was shown to be of the XXXG type and was substituted with galactose and fucose units. The enzyme treatments of the fibres demonstrated that the xylan and xyloglucan polymers in the fibres were not accessible to the enzyme but that material which was entrapped by the cellulose could be released by the hydrolysis of this cellulose.

Cellobiose dehydrogenase (CDH) is an extracellular enzyme produced by various wood-degrading fungi. It oxidizes soluble cellodextrins, mannodextrins and lactose efficiently to their corresponding lactones by a ping-pong mechanism using a wide spectrum of electron acceptors including quinones, phenoxyradicals, Fe(3+), Cu(2+) and triiodide ion. Monosaccharides, maltose and molecular oxygen are poor substrates. CDH that adsorbs strongly and specifically to cellulose carries two prosthetic groups; namely, an FAD and a heme in two different domains that can be separated after limited proteolysis. The FAD-containing fragment carries all known catalytic and cellulose binding properties. One-electron acceptors, like ferricyanide, cytochrome c and phenoxy radicals, are, however, reduced more slowly by the FAD-fragment than by the intact enzyme, suggesting that the function of the heme group is to facilitate one-electron transfer. Non-heme forms of CDH have been found in the culture filtrate of some fungi (probably due to the action of fungal proteases) and were for a long time believed to represent a separate enzyme (cellobiose:quinone oxidoreductase, CBQ). The amino acid sequence of CDH has been determined and no significant homology with other proteins was detected for the heme domain. The FAD-domain sequence belongs to the GMC oxidoreductase family that includes, among others, Aspergillus niger glucose oxidase. The homology is most distinct in regions that correspond to the FAD-binding domain in glucose oxidase. A cellulose-binding domain of the fungal type is present in CDH from Myceliophtore thermophila (Sporotrichum thermophile), but in others an internal sequence rich in aromatic amino acid residues has been suggested to be responsible for the cellulose binding. The biological function of CDH is not fully understood, but recent results support a hydroxyl radical-generating mechanism whereby the radical can degrade and modify cellulose, hemicellulose and lignin. CDH has found technical use in highly selective amperometric biosensors and several other applications have been suggested.