Enzyme and Microbial Technology最新文献

{"title":"Corrigendum to “Influence of a rumen ciliate-derived xylanase on the gut microbiota composition: A potential enzyme for prebiotic applications” [Enzyme Microb. Technol. 190 (2025) 110683]","authors":"Weide Su , Huiying Luo , Gaoxiang Ai , Qipeng Wei , Zhiheng Zou , Xiaolian Chen , Chuanhui Xu , Jiang Chen , Pingwen Xiong , Wenjing Song , Qiongli Song","doi":"10.1016/j.enzmictec.2025.110749","DOIUrl":"10.1016/j.enzmictec.2025.110749","url":null,"abstract":"","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110749"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145039357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of Bacillus subtilis for enhanced p-Coumaric acid production and antimicrobial applications","authors":"Junjie Zhang ,&nbsp;Guodong Zhang ,&nbsp;Wenhu Zhu ,&nbsp;Youzhi Li ,&nbsp;Yutuo Wei ,&nbsp;Xianwei Fan","doi":"10.1016/j.enzmictec.2025.110762","DOIUrl":"10.1016/j.enzmictec.2025.110762","url":null,"abstract":"<div><div><em>p</em>-Coumaric acid (<em>p</em>-CA) is widely utilized in the food, pharmaceutical and other industries, and has traditionally been derived from plant extraction or chemical synthesis. However, <em>p</em>-CA synthesized by the safe <em>B. subtilis</em> remains poorly explored. In this study, we first engineered a recombinant <em>B. subtilis</em> strain (PBK) capable of synthesizing <em>p</em>-Coumaric acid, achieving an initial yield of 3.81 mg L⁻¹. A high-yielding strain PBnprE was then developed through promoter substitution, with a yield reaching 60.92 mg L⁻¹, and the yield of PBnprE was further increased to 304.04 mg L⁻¹ by optimizing fermentation conditions and substrates, showing an 80-fold increase over PBK. The optimized fermentation extract of PBnprE displayed increased antibacterial activity against <em>Staphylococcus aureus</em>, <em>Pseudomonas aeruginosa</em>, and <em>Escherichia coli</em>, alongside enhanced DPPH and ABTS scavenging capabilities. Compared to PBK, the optimized extracts showed 4.81-fold higher in DPPH and 3.47-fold in ABTS scavenging, consistent with improved antioxidant properties driven by the increased presence of <em>p</em>-CA. This study first successfully constructed a high-yield <em>p</em>-CA producing engineered strain in <em>B. subtilis</em>, providing a valuable platform for synthesizing other secondary metabolites.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110762"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145312635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rosmarinic acid production using advanced metabolic engineering strategies","authors":"Beining Wang ,&nbsp;Jintao Lu ,&nbsp;Ruzhe Zhang ,&nbsp;Jung-Kul Lee ,&nbsp;Vipin Chandra Kalia ,&nbsp;Chunjie Gong","doi":"10.1016/j.enzmictec.2025.110748","DOIUrl":"10.1016/j.enzmictec.2025.110748","url":null,"abstract":"<div><div>With improved socio-economic conditions and heightened health awareness, modern consumers now prioritize the nutritional and functional attributes of food over basic satiety. Recently, a high value component, rosmarinic acid synthesized in plants have gained attention as functional food ingredients. Traditional strategy of rosmarinic acid production, including chemical synthesis and plant extraction, are limited by environmental concerns, low yields, and high costs. With the development of biotechnology, metabolic engineering is an alternative strategy for the efficient and cost-effective production of rosmarinic acid. This review focuses on metabolic engineering advances featuring three core strategies: dynamic pathway regulation, cofactor recycling, and microbial co-culture systems. These potential innovations hold great promise for significantly enhancing rosmarinic acid yields. In addition, the review evaluates the economic and technical feasibility of large-scale production, emphasizing the addressing of challenges from traditional production methods.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110748"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145027364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of a thermostable GH6 family cellulase from chaetomium thermophilum exhibiting high cellobiose and ionic liquid tolerance","authors":"Pooja ,&nbsp;Sushant K. Sinha ,&nbsp;Supratim Datta","doi":"10.1016/j.enzmictec.2025.110755","DOIUrl":"10.1016/j.enzmictec.2025.110755","url":null,"abstract":"<div><div>Efficient conversion of lignocellulosic biomass into fermentable sugars requires cellulases that are both thermostable and tolerant to inhibitors such as ionic liquids (ILs). Thermostable enzymes are particularly valuable for industrial applications, as they maintain activity at elevated temperatures for extended periods, improve product yield, and reduce process costs. In this study, we cloned and characterized <em>Ct</em>Cel6C, a GH6 family cellulase, from the thermophilic fungus <em>Thermochaetoides thermophila</em> (<em>Chaetomium thermophilum</em>). <em>Ct</em>Cel6C exhibited a specific activity of 28 U/mg on carboxymethyl cellulose (CMC-Na) at 55 °C and pH 6.0, retaining 90 % of its specific activity after 20 h at optimum pH and temperature (55 °C and pH 6.0). <em>Ct</em>Cel6C demonstrated broad substrate specificity, effectively hydrolyzing oligosaccharides, soluble substrates (CMC and barley β-glucan), and insoluble substrates such as Avicel, consistently producing cellobiose as the sole product and outperforming other GH6 cellobiohydrolases from <em>C. thermophilum</em>. Despite having an open active-site cleft due to a lack of a fifteen-residue stretch in the C-terminal loop, <em>Ct</em>Cel6C retains the biochemical characteristics of a processive cellobiohydrolase rather than those of an endoglucanase from the GH6 family. The enzyme also exhibited high tolerance to cellobiose, glucose, and to 20 % (v/v) of the ionic liquid 1-ethyl-3-methylimidazolium acetate ([C₂mim][OAc]), in both McIlvaine buffer (pH 6.0) and seawater. The remarkable IL tolerance of <em>Ct</em>Cel6C makes it a promising candidate for integration into industrial enzyme cocktails for biomass saccharification.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110755"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145217523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced synthesis of alanyl-glutamine dipeptide via molecular modification and process optimization of α-amino acid ester acyltransferase EAET","authors":"Jinao Wei ,&nbsp;Xiaoxi Liu ,&nbsp;Yifei Gao,&nbsp;Jing-wen Yang,&nbsp;Hong-bin Zhang,&nbsp;Xue-qin Hu","doi":"10.1016/j.enzmictec.2025.110754","DOIUrl":"10.1016/j.enzmictec.2025.110754","url":null,"abstract":"<div><div>This study presents a significant advancement in the enzymatic synthesis of alanyl-glutamine (Ala-Gln) through the development of a high-performance α-ester acyltransferase (EAET) using molecular engineering. We successfully cloned EAET from <em>Bacillus pumilus</em>, which exhibits low sequence identity (&lt;30 %) compared to other reported acyltransferases (AETs). Using AlphaFold3, we constructed a highly reliable structural model of EAET, validated by metrics such as pLDDT, ipTM, and PAE. Through semi-rational design, we introduced the F330Y mutation, which enhanced enzymatic activity by 1.5-fold and extended the half-life at 30°C by 1.3-fold. Molecular dynamics simulations revealed increased flexibility around residue 330 and additional hydrogen bonds in the F330Y mutant, both of which contribute to its improved catalytic efficiency and thermostability. Response surface methodology was employed to optimize enzyme production conditions, resulting in a maximum enzyme activity of 218.20 U/mL[91.3 U/(mL·OD600)]. Systematic optimization of catalytic parameters, including pH, temperature, substrate ratio, concentration, enzyme dosage, and reaction time, enabled an 82.59 % conversion yield at 300 mM substrate concentration. These results not only surpass previous benchmarks but also demonstrate the industrial potential of engineered EAET variants for Ala-Gln synthesis. This study highlights the successful integration of computational tools with molecular engineering to enhance enzyme performance, offering a greener and more sustainable approach for Ala-Gln production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110754"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145205964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A thermostable cellobiose phosphorylase from Thermoclostridium caenicola and its application to in vitro biotransformation","authors":"Haotian Cheng ,&nbsp;Yuan Li ,&nbsp;Yi Zhou ,&nbsp;Shuke Wu ,&nbsp;Yi-Heng P. Job Zhang","doi":"10.1016/j.enzmictec.2025.110753","DOIUrl":"10.1016/j.enzmictec.2025.110753","url":null,"abstract":"<div><div>Cellobiose phosphorylase (CBP) catalyzes the reversible phosphorolysis of cellobiose to glucose and α-D-glucose-1-phosphate. CBPs play an important role in the <em>in vivo</em> enzymatic utilization of cellulosic materials. Here we discovered a novel CBP from <em>Thermoclostridium caenicola</em> (TcCBP), displaying 50–75 % sequence homology with previously reported CBPs. Recombinant TcCBP was well expressed in <em>E. coli</em> BL21(DE3), with a 1.6-fold increase in soluble expression levels over the widely-used <em>Clostridium thermocellum</em> CBP (CtCBP). This enzyme exhibited broad pH adaptability, maintaining substantial activities across pH 4.0–7.5 in the synthetic direction and pH 5.0–7.5 in the phosphorolytic direction. Compared to CtCBP, TcCBP displayed the superior thermostability and a nearly 100-fold improvement in \"Product-to-Enzyme Ratio\" (PE value). The biosynthesis of <em>myo</em>-inositol from cellobiose was conducted by using this CBP along with other four thermophilic enzymes (i.e., phosphoglucomutase, inositol 1-phosphate synthase, inositol monophosphatase, and polyphosphate glucokinase) in one pot without step-by-step addition of enzymes. Approximately 96 mM of <em>myo</em>-inositol was produced from 50 mM of cellobiose. These results indicated that this enzyme could be a potential thermophilic enzyme used for the production of value-added biochemicals by <em>in vitro</em> BioTransformation.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110753"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145109831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation of magnetic xylanase cross-linked aggregates (m-CLXAs) for the hydrolysis of arabinoxylan by mechano-enzymology","authors":"Justin B. Safari ,&nbsp;Lebogang Ramatsui ,&nbsp;Liam Nisbet ,&nbsp;Vincent J. Smith ,&nbsp;Rui W.M. Krause ,&nbsp;Brett I. Pletschke","doi":"10.1016/j.enzmictec.2025.110756","DOIUrl":"10.1016/j.enzmictec.2025.110756","url":null,"abstract":"<div><div>The development of a biocatalyst consisting of immobilised xylanases as cross-linked enzyme aggregates (CLEAs) on magnetic nanoparticles (MNPs) as support was undertaken. MNPs were prepared by a coprecipitation reaction of Fe^3 + and Fe²⁺, followed by surface modification with (3-aminopropyl)-trimethoxysilane using a polycondensation reaction. Magnetic CLEAs were prepared via glutaraldehyde cross-linking after precipitation with acetone, and the physicochemical characterisation of the immobilised enzyme was performed at all stages using several techniques (FTIR, PXRD, TGA, and DLS). A one-factor-at-a-time approach (OFAT) was used to investigate the impact of temperature, time, enzyme: MNP ratio, precipitant, and cross-linking agent to determine their effect on the enzyme’s recovered activity. The results demonstrated that all parameters impacted the immobilisation differently, with the optimised conditions determined as 4°C, 12 h, 10 mg/g, 60 % (v/v) acetone, and 200 mM glutaraldehyde, respectively. The immobilisation did not affect the pH and temperature optima of the enzyme, which were 6.0 and 50–70°C, respectively, for both the free and immobilised enzymes. In contrast, the immobilised enzyme could be reused more than ten times to hydrolyse wheat arabinoxylan, without losing 50 % of its initial activity. Values for <em>V</em>_<em>max</em> and <em>K</em>_<em>m</em> only decreased slightly compared to those obtained for the free enzyme. The thermal inactivation parameters showed that the immobilisation procedure did not adversely affect the enzyme's catalytic properties after immobilisation. Finally, we assessed the immobilised enzyme for its ability to catalyse reactions under mechanochemical conditions (grinding and ageing) and found that the free and immobilised enzymes were active during solvent-free and liquid-assisted grinding (LAG).</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110756"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145198823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Strong alone, weak together: biofilm tensile strength in kangaroo rat burrows","authors":"Duygu Aydin ,&nbsp;Idil Deniz Akin ,&nbsp;Douglas R. Call ,&nbsp;Haluk Beyenal","doi":"10.1016/j.enzmictec.2025.110752","DOIUrl":"10.1016/j.enzmictec.2025.110752","url":null,"abstract":"<div><div>Desert kangaroo rats (<em>Dipodomys deserti</em>) construct burrows that create protected micro-niches favorable to increased microbial activity and biofilm formation. Biofilms within these burrows bind sand particles together, increase the tensile strength of the burrow sand and burrow stability. Our previous work 1) demonstrated that kangaroo rat burrow sand exhibits higher tensile strength than surrounding surface sand due to the cementation by biofilms, and 2) characterized the microbial biofilm communities within kangaroo rat burrow sand and isolated abundant microorganisms. In this study, isolated species are used to quantify tensile strength of pure and mixed biofilms using the microcantilever technique. Mono-species biofilms of <em>Aspergillus tamarii</em> (59.30 ± 4.36 kPa) exhibited the highest tensile strength, while <em>Neobacillus niacini</em> (9.45 ± 1.98 kPa) showed the lowest. Dual-species biofilms displayed synergistic or antagonistic effects, depending on species combinations. Mixing <em>N. niacini</em> with <em>Peribacillus frigoritolerans</em> increased tensile strength to 55.11 ± 2.51 kPa, whereas combining <em>A. tamarii</em> with <em>N. niacini</em> reduced the tensile strength to 18.98 ± 2.54 kPa. Mixing up to five species reduced tensile strength to a minimum value of 2.16 kPa. We conclude that biofilms formed by microbial isolates from burrow sand individually had higher tensile strength, but when all were mixed, the tensile strength decreased, making them weaker.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110752"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145106315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation, expression and characterization of a novel thermo-acid/alkali-stable GH10 xylanase BsXynA from Bacillus safensis L7 and its potential for xylooligosaccharide production and animal feed saccharification","authors":"Ting Zhang,&nbsp;Zhong Cheng,&nbsp;YuMei Fan,&nbsp;YuXin Lan,&nbsp;HuiLan Shu,&nbsp;JinHua Chen,&nbsp;FengCheng Jin,&nbsp;LiYuan Qin,&nbsp;DongPing Feng","doi":"10.1016/j.enzmictec.2025.110735","DOIUrl":"10.1016/j.enzmictec.2025.110735","url":null,"abstract":"<div><div>Xylanases have wide applications in agro-industrial processes. This study reports the discovery and characterization of a novel thermo-acid/alkali-stable GH10 xylanase (BsXynA) from <em>Bacillus safensis</em> L7. The xylanase gene (<em>BsxynA</em>) was cloned and expressed in <em>Escherichia coli</em> BL21 (DE3), yielding a protein of approximately 64 kDa. BsXynA exhibited optimal activity (17.33 U/mg) on beechwood xylan at pH 6.0 and 60°C. Moreover, BsXynA exhibited remarkable thermo-acid/alkali stability, retaining over 60 % activity at pH 5.0–8.0 after 60 min at 60°C and over 80 % activity after 14 days at 4°C within pH 6.0–9.0. Additionally, the enzyme tolerated 50°C and various chemicals, with a half-life of over 16 days. It was activated by K⁺, Na⁺, Ca²⁺, Ba²⁺, and Mg²⁺ ions but inhibited by Zn²⁺, Cu²⁺, and SDS. BsXynA hydrolyzed various xylans but not glucose-based polysaccharides. <em>K</em>_m, <em>V</em>_max, <em>k</em>_cat, and <em>k</em>_cat/<em>K</em>_m for beechwood xylan hydrolysis were found to be 6.61 mg/mL, 24.24 µmol·min⁻¹·mg⁻¹, 15.71 s⁻¹, and 2.38 mL·s⁻¹·mg⁻¹ respectively. Thin-layer chromatography (TLC) analysis showed that BsXynA is an endo-type xylanase, which hydrolyzes beechwood xylan to produce mainly xylobiose (X2) and xylotetraose (X4), with no xylose detected. Furthermore, BsXynA improved animal feed saccharification, making it a promising biocatalyst for biotechnological applications.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110735"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144841943","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and recombinant expression of a novel defluorinase from Rhodococcus jostii RHA1, for defluorination and biotransformation of the PFAS compound 6:2 fluorotelomer carboxylic acid","authors":"Eustace Y. Fernando","doi":"10.1016/j.enzmictec.2025.110724","DOIUrl":"10.1016/j.enzmictec.2025.110724","url":null,"abstract":"<div><div>Poly and per fluorinated substances (PFAS) are emerging contaminants of concern that are thought to be involved in causing numerous adverse health effects, such as immunosuppression, increased chance of cancer development, and altered levels of hepatic enzyme levels in humans. However, PFAS are considered highly persistent and resistant to biodegradation given the fact that the C-F bond can have a bond dissociation energy of up to 544 kJ/mol. Though many studies have reported PFAS biodefluorination by bacterial isolates and microbial communities, little is known regarding the molecular foundations for biodefluorination. In this study, a novel defluorinase was identified, that is responsible for the biodefluorination of 6:2 fluorotelomer carboxylic acid (6:2 FTCA) in <em>R</em>.<em>jostii</em> RHA1 using the combination of transposome-based insertional mutagenesis and heterologous expression. From a library of 417 <em>R</em>.<em>jostii</em> RHA1. mutants, 3 individual mutants lost their ability for defluorination when they were exposed to 6:2 FTCA (mutant # 15, 32 and 38 – Table S2). The disruption of the genetic locus in all 3 non-defluorinating mutants was identified coding for a putative MhPC superfamily protein. The MhPC superfamily of proteins is known to harbor other proteins such as fluoroacetate dehalogenase (UniProt - Q6NAM1) that are capable of –C-F bond cleavage. This identified gene was cloned into the heterologous expression host <em>M. smegmatis</em> MC²-155. After induction, the <em>M. smegmatis</em> MC²-155 transformant exhibited the ability to defluorinate 6:2 FTCA at a rate of 13 µmol/h (V_max = 80.9 µmol/min and K_m = 5.04 mM in Michaelis-Menten models). In contrast, defluorination was not observed in either abiotic or biotic controls. Further characterization of the novel defluorinase indicated that it could moderately defluorinate the unsaturated PFAS compound 6:2 FTCUA (4.9 µmol/h fluoride) and minimally defluorinate 5:2 sFTOH (1.3 µmol/h fluoride). The novel defluorinase indicated a maximal specific activity of 12.9 ± 1.9 µmol F/hr/g protein, against its primary PFAS substrate, 6:2 FTCA. However, it showed no activity with 5:3 FTCA or sulfonated PFAS compounds such as 6:2 FTS and 8:2 FTS. The wild-type <em>Rhodococcus</em> could defluorinate 6:2 FTCA at a rate of 2.2 µmol/h. The discovery of this MhPC class novel defluorinase in WT <em>R</em>.<em>jostii</em> RHA1. has substantial value since it is responsible for the critical step that initiates defluorination of PFAS compounds such as 6:2 FTCA.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110724"},"PeriodicalIF":3.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144702367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}