Enzyme and Microbial Technology最新文献

{"title":"Establishing a novel pathway for the biosynthesis of nicotinamide mononucleotide.","authors":"Rongchen Feng, Ziting Yan, Guoguang Wei, Chaoqiang Wu, Feifei Chen, Alei Zhang, Sheng Xu, Xin Wang, Kequan Chen","doi":"10.1016/j.enzmictec.2025.110633","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2025.110633","url":null,"abstract":"<p><p>Nicotinamide mononucleotide (NMN) is a pivotal molecule within the realm of metabolic health, serving as a precursor to nicotinamide adenine dinucleotide (NAD⁺), a critical coenzyme in cellular energy metabolism. In recent years, the biological production of NMN has garnered significant interest. In this study, we developed the novel NRK-dependent synthesis routes for NMN production. Two strategies were designed to supply D-ribose-1-phosphate (R-1-P): (1) phosphorylation of exogenous D-ribose to ribose-5-phosphate (R-5-P) using engineered ribokinase (RK), followed by isomerization to R-1-P; (2) R-5-P synthesis from glucose through the pentose phosphate pathway. An optimized in vitro multi-enzyme cascade (XapA/PNP/NRK, PPM, NRK) identified NRK as the most efficient catalyst for NMN biosynthesis from D-ribose and niacinamide. In Escherichia coli, overexpression of this cascade, knockout of competing pathways, and secretion enhancement via a pelB signal peptide-fused PnuC transporter achieved an NMN titer of 62.0 mg L^-¹ .This work provides a viable alternative for the biosynthesis of NMN.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":" ","pages":"110633"},"PeriodicalIF":3.4,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143709236","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":"Characterization of a novel short-chain dehydrogenase from Sugiyamaella lignohabitans and its application in the degradation of patulin","authors":"Wei Xu ,&nbsp;Jiayi Yao ,&nbsp;Chenyu Song ,&nbsp;Ting Xue ,&nbsp;Wanmeng Mu","doi":"10.1016/j.enzmictec.2025.110630","DOIUrl":"10.1016/j.enzmictec.2025.110630","url":null,"abstract":"<div><div>Patulin (PAT) is a widespread and hazardous fungal toxin that is commonly found in fruits and grain crops, threatening global food safety. Increasing attention has been focused on biological approaches for PAT decontamination in recent years. This study successfully identified a short-chain dehydrogenase with PAT-degrading activity derived from <em>Sugiyamaella lignohabitans</em> and made a preliminary characterization of the recombinant enzyme <em>Suli</em>-SDR. The <em>Suli</em>-SDR displayed the optimal activity at pH 7.0 and 80 °C, indicating a good activity when applied in the pasteurization procedure of the apple juice. <em>Suli</em>-SDR with a final concentration of 1 μg/mL could degrade 88 % of PAT (50 μg/mL) within 12 h, revealing its superior degradation performance. After measuring the parameters related to the quality of apple juice, including vitamin C, titratable acids, total phenols, etc., it was found that there was no significant effect on the parameters of apple juice after enzymatic treatment by <em>Suli</em>-SDR. In a short summary, this study evaluated the catalytic ability of <em>Suli</em>-SDR and explored the feasibility of enzyme on PAT control in practical production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110630"},"PeriodicalIF":3.4,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143680587","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":"In-vitro optimization and active-site mutagenesis of CYP105D18 peroxygenase enhance the production of indigo","authors":"Bashu Dev Pardhe ,&nbsp;HyunA Park ,&nbsp;Prakash Paudel ,&nbsp;Jaeho Jeong ,&nbsp;Tae-Jin Oh ,&nbsp;Kwon-Young Choi ,&nbsp;Jungoh Ahn","doi":"10.1016/j.enzmictec.2025.110634","DOIUrl":"10.1016/j.enzmictec.2025.110634","url":null,"abstract":"<div><div>Practical implementation of efficient biocatalysts for large-scale production of indigo remains challenging. Microbial cytochrome P450s may be useful for indigo production, but this has been rarely reported. We discovered that CYP105D18 catalysed H₂O₂-mediated C-3 hydroxylation of indole to synthesize indigo. A cell-free lysate from <em>Escherichia coli</em> containing CYP105D18 peroxygenase obtained after cell disruption was optimized for <em>in vitro</em> reaction. Next, 250 µM hydroxylamine was added to the cell-free lysate to inhibit other H₂O₂-utilizing enzymes that interfere with the CYP105D18 function. Furthermore, the active-site residues of CYP105D18, namely L87, A235, A282, and I386, involved in indole binding were mutated. L87F resulted in an approximately 12-fold increase in CYP105D18 activity. The catalytic efficiencies of the wild-type and L87F mutant were 0.01 and 0.12 mM⁻¹min⁻¹, respectively. Fed-batch fermentation using enriched autoinduction medium was used for higher production of <em>E. coli</em> cells containing CYP105D18 peroxygenase. The Cell-free lysate of disrupted cells yielded 710 mg/L of indigo in 20 min. This represents a simple enzymatic approach for indigo biosynthesis using cell-free lysate of <em>E. coli</em> overexpressing CYP105D18, H₂O₂, and catalase inhibitor without the need for multi enzyme systems and expensive cofactors. This single-enzyme system, used in a rapid process for indigo formation, could serve as an efficient approach for commercial bio-indigo production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110634"},"PeriodicalIF":3.4,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143680646","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 novel GH12 xyloglucanase from the white rot fungus Abortiporus biennis, synergistically enhances lignocellulose saccharification by commercial cellulases","authors":"Despoina Panoraia Bakouli ,&nbsp;Elisavet Pedi ,&nbsp;Nikolaos Labrou ,&nbsp;Evangelos Topakas ,&nbsp;Anastasia Zerva","doi":"10.1016/j.enzmictec.2025.110628","DOIUrl":"10.1016/j.enzmictec.2025.110628","url":null,"abstract":"<div><div>Xyloglucan is a complex, highly substituted plant biomass polysaccharide, which is largely overlooked in the design of enzyme cocktails for lignocellulose saccharification, due to its presence in specific plant tissues only, and its low content. Thus, the microbial mechanisms for its degradation have not been thoroughly studied. However, in the frame of the biorefinery concept, xyloglucan monomers also have to be utilized for the design of efficient bioprocesses. Moreover, in plant tissues, xyloglucan often covers cellulose fibrils, impeding the access of cellulases. In order to shed light on the enzymatic degradation of xyloglucan, a novel GH12 family xyloglucanase was studied, from the basidiomycete <em>Abortiporus biennis</em>. The enzyme was heterologously produced in <em>Pichia pastoris</em>, purified and characterized. <em>Abi</em>Xeg12a is a 28 kDa glycoprotein, with relatively strict substrate specificity, since it is only active in xyloglucan and β-glucan. The main hydrolysis products are the oligomers XXXG, XLXG/XXLG, XLLG and the optimum activity conditions are pH 4.5 and 55 °C. The enzyme contributes to the saccharification of corn bran and apple pulp by a commercial cellulase preparation, increasing the release of reducing sugars by up to 39 % and 18 %, respectively, while the addition of <em>Abi</em>Xeg12a can minimize the enzyme load of the reaction, at least for apple pulp, without loss in reducing sugar yield. Overall, the importance of xyloglucanases on the saccharification of xyloglucan-containing substrates was demonstrated in this study. The results could contribute to the design of more efficient, tailor-made enzyme cocktails for the saccharification and subsequent valorization of lignocellulose.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110628"},"PeriodicalIF":3.4,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143642329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A metabolic-engineering framework approach via fed-batch fermentation for enhancing glucaric acid production in Komagataella phaffii","authors":"Jayachandran Krishna ,&nbsp;Kabilan Subash Chandra Bose ,&nbsp;Sindhu Varadharaj ,&nbsp;Meenakshisundaram Sankaranarayanan","doi":"10.1016/j.enzmictec.2025.110627","DOIUrl":"10.1016/j.enzmictec.2025.110627","url":null,"abstract":"<div><div>Glucaric acid (D-saccharic acid) is an organic compound belonging to glucuronic acid derivatives, whose commercial synthesis involves the use of hazardous solvents. Biosynthetic production in <em>Saccharomyces cerevisiae</em> has limitations, such as ethanolic fermentation, redox strategy limitations, and low pH toxicity. <em>Komagataella phaffii</em> (<em>K. phaffii</em>) formly known <em>Pichia pastoris</em>, an alternative and robust engineerable organism, is a promising biotransformation agent for glucaric acid production. However, <em>K. phaffii</em> lacks native biosynthetic pathways for glucaric acid synthesis at the industrial scale. There is no proof-of-concept glucaric acid production system. Therefore, gene expression profiling-based metabolic engineering of glucaric acid producing gene cassette was performed using in-fusion cloning. Product production was enhanced using fed-batch fermentation of the key metabolite, myo-inositol; this improved the yield of glucaric acid. The expression was optimized through cofactor recycling and codon optimization for the UDH gene. Fed-batch fermentation with mixed supplementation (Myo-inositol + Monosodium glutamate) as substrate in engineered <em>K. phaffii</em> (X33-GA) enhanced glucaric acid synthesis to 17.6 g/L. In addition, we present simple HPLC and LC-MS techniques for quantifying glucaric acid and its precursors in the fermentation samples. The proof-of-concept results from both shake flask and bioreactor studies provide a unique perspective on sustainable, cost-effective, and green technological alternatives for glucaric acid synthesis.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110627"},"PeriodicalIF":3.4,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143642330","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":"Synergy of GH67 and GH115 α-1,2-glucuronidases with Penicillium subrubescens endoxylanases to stimulate xylooligosaccharide production","authors":"Xinxin Li ,&nbsp;Lanyu Li ,&nbsp;Alessia Manassero ,&nbsp;Astrid Müller ,&nbsp;Sumitha K. Reddy ,&nbsp;Mirjam A. Kabel ,&nbsp;Ronald P. de Vries ,&nbsp;Peicheng Sun","doi":"10.1016/j.enzmictec.2025.110629","DOIUrl":"10.1016/j.enzmictec.2025.110629","url":null,"abstract":"<div><div>A primary substitution of the plant cell wall hemicellulosic polysaccharide xylan is (4-<em>O</em>-methyl-)<span>d</span>-glucuronic acid, which hinders the endoxylanases (XLNs) degradation of xylan for the production of valuable xylooligosaccharides (XOS). In this context, α-1,2-glucuronidase (AGU) plays a critical role in hydrolyzing the α-(1→2)-glycosidic linkages between 4-<em>O</em>-methyl-<span>d</span>-glucuronic acid and xylosyl residues in xylan, thereby enhancing XOS production by XLNs. However, AGUs have been relatively poorly studied, and insufficient and incomplete data on their biochemical properties, substrate specificity, and product profiling has limited their application. Here, we cloned, heterologously produced, purified and functionally characterized an AGU from <em>Aspergillus niger</em> (<em>An</em>AguA) and another AGU from <em>Penicillium subrubescens</em> (<em>Ps</em>AguB), belonging to Glycoside Hydrolase family 67 (GH67) and 115 (GH115), respectively, in the Carbohydrate-Active enZyme database. Results showed that neither AGU released 4-<em>O</em>-methyl-<span>d</span>-glucuronic acid from polymeric beech wood glucuronoxylan (BeWX). However, we found that from BeWX pre-digested with GH10 or GH11 XLNs from <em>P. subrubescens</em> (<em>Ps</em>XlnA and <em>Ps</em>XlnF, respectively), <em>An</em>AguA released 4-<em>O</em>-methyl-<span>d</span>-glucuronic acid only from the non-reducing end of glucuronoxylan oligosaccharide, whereas <em>Ps</em>AguB released 4-<em>O</em>-methyl-<span>d</span>-glucuronic acid from glucuronoxylan oligosaccharides regardless of the xylosyl substitution position. Furthermore, we demonstrated that enhancement of XOS release by adding AGUs to various combinations of GH10 (<em>Ps</em>XlnA–C) and GH11 (<em>Ps</em>XlnD–F, <em>Ps</em>XlnH–I) XLNs from <em>P. subrubescens</em> varied based on the AGU-XLN combination. The combination of <em>An</em>AguA with <em>Ps</em>XlnA was the most effective, achieving at least a 3-fold increase in the release of XOS with a degree of polymerization of 5–7 compared to using <em>Ps</em>XlnA alone.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110629"},"PeriodicalIF":3.4,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143637135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acetic acid production from corn straw via enzymatic degradation using putative acetyl esterase from the metagenome assembled genome","authors":"Hao Chen ,&nbsp;Zhiwei Zeng ,&nbsp;Rup Lal ,&nbsp;Jie Wu ,&nbsp;Jia Chen ,&nbsp;Mei Li ,&nbsp;Lulu Cao ,&nbsp;Xiqiang Liu ,&nbsp;Ruzhe Zhang ,&nbsp;Chunjie Gong","doi":"10.1016/j.enzmictec.2025.110619","DOIUrl":"10.1016/j.enzmictec.2025.110619","url":null,"abstract":"<div><div>Acetic acid production from corn straw by enzyme catalysis shows its application value in food industry. In this study, a gene encoding for a putative acetyl esterase derived from <em>Sphingobacterium soilsilvae</em> Em02 was discovered in metagenome assembled genome. The gene was expressed in <em>Escherichia coli</em> BL21 to obtain enzyme with a molecular mass of 38.8 kDa. P-Nitrophenyl acetate was used as a substrate to determine the enzyme activity. The enzyme demonstrated optimal activity under conditions of 40 °C and a neutral pH of 7.0. Under optimal conditions, 17.58 mg of acetic acid was obtained using the enzyme from 50 mg corn straw pretreated with amylase. The acetyl esterase derived from <em>Sphingobacterium soilsilvae</em> Em02, demonstrates significant potential for biotechnological applications, particularly in biomass degradation.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"187 ","pages":"Article 110619"},"PeriodicalIF":3.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143576748","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":"Recombinant alginate lyases and mannitol dehydrogenase enhance hydrolysis of macroalgal carbohydrates","authors":"Dominique F. Rocher,&nbsp;Marthinus W. Myburgh,&nbsp;Marinda Viljoen-Bloom,&nbsp;Rosemary A. Cripwell","doi":"10.1016/j.enzmictec.2025.110618","DOIUrl":"10.1016/j.enzmictec.2025.110618","url":null,"abstract":"<div><div>Brown macroalgae are a promising source for bioethanol production, primarily due to their high carbohydrate, low lignin and high moisture content. Bioconversion of macroalgae to ethanol requires a yeast, such as <em>Saccharomyces cerevisiae</em>, that can hydrolyse the macroalgal carbohydrates, namely laminarin, mannitol and alginate. In this study, the mannitol dehydrogenase (MDH) genes from <em>Aspergillus fumigatus</em> (<em>AfMDH</em>) and <em>Talaromyces islandicus</em> (<em>TiMDH</em>), and the alginate lyase (AL) genes from <em>Sphingomonas</em> sp. (<em>SpxAL</em> and <em>SpeAL</em>) and <em>Talaromyces emersonii</em> (<em>TeeAL</em>) were expressed in the laboratory strain, <em>S. cerevisiae</em> Y294. Co-cultures of a laminarinase-producing yeast, Y294[Relam1/Tvlam1] and yeasts expressing mannitol dehydrogenases and alginate lyases were evaluated for the consolidated bioprocessing of the major carbohydrates in brown macroalgae. Laminarin and mannitol were targeted for ethanol production, while alginate was depolymerised to expose mannitol. A co-culture of <em>S. cerevisiae</em> Y294[Relam1/Tvlam1], [AfMDH] and [TeeAL/SpxAL] strains produced 10.30 g/L ethanol from <em>Ecklonia maxima</em>, representing a 98 % carbon conversion (based on the laminarin and mannitol content). A strain expressing both endo- and exo-alginate lyase improved the ethanol yield by 42.28 % compared to strains expressing only laminarinase- and mannitol dehydrogenase. Scanning electron microscopy further revealed that co-cultures containing laminarinase, MDH, and AL enzymes promoted significant physical degradation and increased porosity in macroalgal substrates, suggesting enhanced alginate hydrolysis and improved enzyme accessibility. This is the first report on the simultaneous hydrolysis of mannitol, alginate and laminarin with recombinant enzymes during macroalgal fermentation. The results demonstrate significant progress towards exploiting brown macroalgae for bioconversion to ethanol and high-value products.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"186 ","pages":"Article 110618"},"PeriodicalIF":3.4,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143551515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synergy between processive cellulases in Ruminoccocus albus","authors":"Alem Storani ,&nbsp;Alberto A. Iglesias,&nbsp;Sergio A. Guerrero","doi":"10.1016/j.enzmictec.2025.110610","DOIUrl":"10.1016/j.enzmictec.2025.110610","url":null,"abstract":"<div><div>Endoglucanases (EGs), cellobiohydrolases (CBHs), and β-glucosidases are essential components in enzymatic degradation of cellulose. We analyzed the glycosyl hydrolases from families GH5 and GH48 from <em>Ruminococcus albus</em> 8 (<em>Ral</em>Cel5G and <em>Ral</em>Cel48A). Both enzymes feature a catalytic motif and a carbohydrate binding domain from family 37 (CBM37). <em>Ral</em>Cel5G also exhibited a second CBM37 with lower similarity. As a result, <em>Ral</em>Cel5G showed higher binding affinity toward insoluble substrates and broader recognition capacity. Kinetic characterization using different cellulosic substrates and reaction product analysis confirmed <em>Ral</em>Cel5G as a processive EG while <em>Ral</em>Cel48A is a CBH. Interestingly, we found a synergistic effect on their activity at a low EG to CBH ratio, despite the processive activity of <em>Ral</em>Cel5G. Furthermore, the lignocellulose degradation capacity was improved by supplementing the cellulases with hemicellulase <em>Ral</em>Xyn10A. These results provide valuable information about the interaction between processive EG and conventional CBH, necessary for the rational design of enzyme cocktails for optimized biomass processing.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"186 ","pages":"Article 110610"},"PeriodicalIF":3.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143455005","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":"Characterization of keratinase from Chryseobacterium camelliae Dolsongi-HT1 and efficacy on skin exfoliation","authors":"Eun-Mi Kim,&nbsp;Soojung Oh,&nbsp;Hyeongwon Choi,&nbsp;Won-Seok Park","doi":"10.1016/j.enzmictec.2025.110605","DOIUrl":"10.1016/j.enzmictec.2025.110605","url":null,"abstract":"<div><div>Keratin is the outermost layer that protects our skin and has an appropriate turnover cycle. With age, the keratin turnover cycle begins to dysfunction. To overcome this issue, we artificially remove dead skin cells. In this study, we attempted to screen enzymes that could be useful in the cosmetics industry to develop enzymes suitable for the enzyme-based method, a mild exfoliation method that does not damage the skin. <em>Chryseobacterium camelliae</em> Dolsongi-HT1 with keratinolytic activity was isolated from green tea leaves (sourced from the Dolsongi tea garden, Jeju Island). The keratinolytic activity of <em>C. camelliae</em> Dolsongi-HT1 was detected in the culture media, indicating that the target keratinolytic enzyme is a secreted protein. Keratinolytic activity was demonstrated using forearm skin keratin and reconstituted human skin models. The enzyme from <em>C. camelliae</em> Dolsng-HT1 (HT1) could efficiently decompose human skin keratin. Moreover, experiments using the reconstituted human skin model demonstrated that HT1 is efficient in exfoliating the outermost stratum corneum. Compared with the popularly used chemical exfoliation method, enzymatic exfoliation using HT1 was less abrasive and did not damage the epidermal layer. Keratinolytic enzyme was identified using protein purification and mass spectrometry. The identified enzyme (iHT1) was expressed in the <em>Bacillus subtilis</em> RIK 1285 secretory protein expression system. The iHT1 enzyme showed high activity over a wide temperature range (30–60 °C), with the highest activity at 30 °C. The optimum pH for the activity of iHT was pH8.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"186 ","pages":"Article 110605"},"PeriodicalIF":3.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143429904","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}