Enzyme and Microbial Technology最新文献

{"title":"Homologous expression and characterization of Coniochaeta ligniaria glycoside hydrolase family 115 α-glucuronidase","authors":"Michael J. Bowman,&nbsp;Nancy N. Nichols","doi":"10.1016/j.enzmictec.2025.110750","DOIUrl":"10.1016/j.enzmictec.2025.110750","url":null,"abstract":"<div><div>Generation of fermentable sugars from biomass is necessary for microbial production of bioproducts. Due to its inherent complexity, xylan hydrolysis to monosaccharides requires several different enzymatic activities. To completely depolymerize glucuronoarabinoxylan from biomass, α-glucuronidase activity is necessary. The ascomycete fungus <em>Coniochaeta ligniaria</em> has the ability to degrade xylan and is an unutilized source of biomass degrading enzymes. Therefore, a gene encoding a putative <em>Coniochaeta ligniaria</em> GH115 α-glucuronidase was cloned and expressed homologously in <em>Coniochaeta ligniaria</em> as a native secreted protein<em>.</em> Culture supernatants were concentrated and purified by a combination of ultrafiltration, anion-exchange, and size-exclusion chromatography. The purified protein behaved as dominantly dimeric complexes as determined by size-exclusion chromatography. The expressed protein liberated: 4-<em>O</em>-methyl glucuronic acid from beech xylan, birch xylan, and beech-derived glucuronoxylooligosaccharides as the sole product of hydrolysis; and 4-<em>O</em>-methyl glucuronic acid and glucuronic acid from oat spelt xylan. The expressed α-1,2-glucuronidase had greater activity on glucuronoxylooligosaccharides than on full length beech xylan. The expressed α-1,2-glucuronidase, herein designated <em>Cl</em>Agu115, had <em>K</em>_<em>m</em> values of: 1.3 mM; 1.2 mM; and 1.0 mM for beech xylan, GH10-hydrolyzed glucuronooligosaccharides, and GH11-hydrolyzed glucuronooligosaccharides, respectively. The measured kinetic constants show that the enzyme prefers an oligosaccharide substrate with the 4-<em>O</em>-methyl glucuronic acid on the non-reducing end. The enzyme had activity between pH 3.0–6.0 and temperatures 10°C-60°C, with optima at pH 4.3 and 40°C. The expression and characterization of <em>Cl</em>Agu115 expands the repertoire of fungal GH115 enzymes for use in biomass conversion.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110750"},"PeriodicalIF":3.7,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046785","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":"2025-09-08","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":"Innovative nanosensing methods for lysozyme identification","authors":"Mohammad Darvishi ,&nbsp;Seyyed Mohammad Mousavinia ,&nbsp;Reza Soleimany ,&nbsp;Mohammad Mahdi Heidari ,&nbsp;Mohsen Mohammadi ,&nbsp;Samad Rastmanesh ,&nbsp;Ahmad Mobed","doi":"10.1016/j.enzmictec.2025.110745","DOIUrl":"10.1016/j.enzmictec.2025.110745","url":null,"abstract":"<div><div>Lysozyme (LYZ) is a critical enzyme recognized for its significant antimicrobial properties, playing an integral role in the immune response and being implicated in various diseases, including infections and inflammatory conditions. Traditional detection methods for LYZ, such as enzyme-linked immunosorbent assays (ELISA) and spectrophotometric techniques, often encounter limitations regarding sensitivity, specificity, and time efficiency. In light of these challenges, there has been substantial advancement in the development of novel biosensor technologies over the past two decades, particularly those that incorporate nanomaterials. These innovative biosensors demonstrate enhanced performance, facilitating rapid and accurate detection of LYZ at low concentrations. This paper aims to provide a comprehensive overview of recently developed biosensors specifically for LYZ, highlighting their design, functionality, and applications in clinical diagnostics and research. Unlike previous reviews, we place a distinct emphasis on the clinical importance of LYZ and its role in various diseases, thereby elucidating its significance in health and disease contexts. Additionally, we will explore the implications of advanced detection methods, particularly those utilizing nanomaterials, for enhancing our understanding of LYZ-related pathologies. By focusing on nanoscale detection techniques, which have not been thoroughly addressed in existing literature, we underscore the transformative potential of these biosensors in LYZ detection. This approach not only contributes to improved disease management but also informs therapeutic strategies, setting our review apart from prior works.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110745"},"PeriodicalIF":3.7,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144989476","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, cloning, and characterization of a novel GH5 cellulase from yak rumen metagenome for enhanced lignocellulose hydrolysis in biofuel production and ruminant feed utilization","authors":"Ibrahim Bature ,&nbsp;Zeyi Liang ,&nbsp;Wu Xiaohu ,&nbsp;Feng Yang ,&nbsp;Yayuan Yang ,&nbsp;Pengcheng Dong ,&nbsp;Xuezhi Ding","doi":"10.1016/j.enzmictec.2025.110737","DOIUrl":"10.1016/j.enzmictec.2025.110737","url":null,"abstract":"<div><div>Lignocellulosic biomass is a promising feedstock for biofuel production, but its complex structure, including cellulose and hemicellulose, challenges efficient enzymatic hydrolysis. <em>CelyA</em>, a novel cellulase from the yak rumen, has potential for improving biomass degradation and biofuel production. The <em>CelyA</em> gene was cloned, expressed, and purified. Biochemical characterization included assessments of pH, temperature, and salt tolerance. The enzyme's substrate specificity was tested on crystalline cellulose, CMC, and other polysaccharides. <em>In vitro</em> rumen fermentation was performed to evaluate its effect on fiber digestibility and microbial community composition. Biofuel production was tested by hydrolyzing maize, rice, and wheat straw. <em>CelyA</em> demonstrated optimal activity at pH 6.0 and 40.0 °C and maintained moderate stability across a wide pH range (3.0–12.0), retaining measurable activity even under strongly acidic and alkaline conditions. The enzyme demonstrated excellent salt tolerance, retaining 91.0 % activity in 1.0 M NaCl. <em>CelyA</em> efficiently degraded maize straw in hydrolysis assays, producing 7.2 µmol/L of reducing sugars. <em>In vitro</em> rumen fermentation with <em>CelyA</em> increased fiber digestibility by 8.3 % for maize straw, 14.5 % for rice straw, and 2.7 % for wheat straw. Gas production also increased significantly, with maize straw showing a 91.3 % increase. 16S rRNA sequencing revealed selective enrichment of <em>Ruminococcus</em> and <em>Prevotella</em>, key cellulolytic microbes. <em>CelyA</em> demonstrates strong potential for biofuel production, efficiently hydrolyzing lignocellulosic biomass and enhancing ruminal fiber digestibility. Its stability, salt tolerance, and substrate specificity make it a valuable enzyme for biofuel production and livestock feed optimization.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110737"},"PeriodicalIF":3.7,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144907276","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":"Developing a urea biosensor and safe blood cleaning method utilizing a novel high throughput La-doped CeO2 nanosized artificial urease with high biocompatibility and enzyme-like activity","authors":"Naushad Ahmad ,&nbsp;Ashok Kumar Bishoyi ,&nbsp;Suhas Ballal ,&nbsp;Aman Shankhyan ,&nbsp;Shaker Al-Hasnaawei ,&nbsp;Karthikeyan Jayabalan ,&nbsp;Laxmidhar Maharana ,&nbsp;Madan Lal","doi":"10.1016/j.enzmictec.2025.110736","DOIUrl":"10.1016/j.enzmictec.2025.110736","url":null,"abstract":"<div><div>Herein, a urea biosensor and safe blood cleaning method utilizing a novel high throughput La-doped CeO₂ nanosized artificial urease with high biocompatibility and enzyme-like activity were developed. The urease-like activity, stability composition, morphological characteristics, size, biocompatibility, and crystalline characteristics of the artificial urease were assessed. Considering its high urease-like activity, it is applied for both urea biosensing and safe blood cleaning. The effective factors on the sensing conditions were optimized, providing a wide linear range over 1–10 µM and a low detection limit of 0.5 µM. The repeatability and reproducibility assessments revealed a %RSD of 1.7 % and 2.7 %, in order. The selectivity studies and real blood analysis revealed highly selective, accurate, and reliable determination of urea in biosamples using the developed method, providing a %recovery of 97.8–99.5 % with a low %RSD range of 1.4–3.3 %. Besides, the safe blood cleaning experiments were also performed by urea mineralization from blood. The results exhibited a high blood cleaning yield of 99.5 % at a short time of 30 min, revealing 1.8-fold higher cleaning efficiency for the artificial urease compared to native urease. The reusability studies showed that the blood cleaning yield was saved for about 10 cycles and then it decreased by increasing the operational cycles, reaching 78.6 % after 17 cycles. The shelf-life of the urease was also evaluated within 30 days, revealing a high shelf-stability for the as-prepared nanosized artificial urease. Consequentially, the nanosized artificial urease can be practically applied for both urea detection and blood cleaning in the real world.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110736"},"PeriodicalIF":3.7,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864722","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":"Breaking the CREB–CBP alliance: Progress, challenges, and therapeutic promise of small-molecule and peptide disruptors of the pKID–KIX interaction","authors":"Hassan A. Rudayni","doi":"10.1016/j.enzmictec.2025.110734","DOIUrl":"10.1016/j.enzmictec.2025.110734","url":null,"abstract":"<div><div>The kinase-inducible domain (pKID) of the transcription factor CREB engages the KIX domain of the co-activator CBP/p300 to drive signal-dependent gene expression that underpins cell proliferation, metabolism and survival. Aberrant CREB–CBP signaling is now implicated in numerous cancers and metabolic disorders, yet until recently the shallow, dynamic pKID–KIX interface was viewed as chemically intractable. This review integrates two decades of progress that overturns that paradigm. We first dissect the structural and allosteric features of KIX that enable coupled folding-and-binding of pKID and reveal hidden ligandable pockets. We then survey the expanding inhibitor repertoire—from early micromolar naphthols (KG-501) through nanomolar naphthamides (666−15) and orally tractable pro-drugs, to high-affinity stapled and D-peptide mimetics—highlighting the assays, structure–activity relationships and pharmacokinetic optimization that have driven each advance. Biophysical and computational insights, including ¹⁹F NMR ligandability maps and millisecond-scale molecular-dynamics trajectories, are shown to guide next-generation design and machine-learning pipelines. Pre-clinical data demonstrate that disrupting CREB–CBP selectively suppresses tumor growth with favorable tolerability, and we outline opportunities for combination therapies, degrader strategies and indication expansion into metabolic and neurocognitive disease. Collectively, these findings position the CREB–CBP interaction as a tractable, multi-modal drug target poised for first-in-human exploration.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110734"},"PeriodicalIF":3.7,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144841939","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":"2025-08-13","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":"Ethyl lactate synthesis in organic media using a magnetic supported CALB","authors":"Paula Nicolás ,&nbsp;Verónica L. Lassalle ,&nbsp;María Luján Ferreira","doi":"10.1016/j.enzmictec.2025.110732","DOIUrl":"10.1016/j.enzmictec.2025.110732","url":null,"abstract":"<div><div>The growing demand for sustainable chemical processes has spurred interest in enzymatic synthesis, particularly for valuable compounds like ethyl lactate. Traditional chemical methods often suffer from drawbacks, highlighting the potential of enzymatic catalysis using immobilized lipases. This study evaluated the performance of magnetic biocatalyst, prepared by immobilizing <em>Candida antarctica</em> lipase B (CALB) on magnetic nanoparticles, for the batch synthesis of ethyl lactate in hexane. Initial experiments using free CALB and commercial Novozym435 proved problematic due to enzyme denaturation and support instability, respectively. While titration-based methods for monitoring the reaction were found to be unreliable due to lactic acid's complex behavior in the reaction medium, titratable acidity reduction suggested an optimal lactic acid to ethanol molar ratio of 1/10. Subsequent HPLC analysis revealed that the magnetic biocatalyst maintained a consistent conversion (%) at higher lactic acid concentrations (up to 17 mg/mL at 45°C, with conversion above 60 % in 5 h), demonstrating its potential for processing larger amounts of substrate. The initial reaction rate was estimated to be 3.8 mM/h. The study also identified experimental challenges in accurate lactic acid quantification and potential catalyst degradation. In conclusion, the magnetic CALB biocatalyst shows promising activity and stability for ethyl lactate synthesis, especially at higher substrate loads, paving the way for further optimization and application in sustainable production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110732"},"PeriodicalIF":3.7,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144828379","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":"Elucidation of biochemical attributes and enzymatic activity of agarase from Saccharophagus degradans 2–40","authors":"Anoth Maharjan ,&nbsp;Beom Soo Kim","doi":"10.1016/j.enzmictec.2025.110733","DOIUrl":"10.1016/j.enzmictec.2025.110733","url":null,"abstract":"<div><div><em>Saccharophagus degradans</em> 2–40 exhibits agarolytic activity, effectively degrading agar into galactose. Both endo- and exo-agarase, as well as neoagarobiose hydrolase (NABH), play important roles in agar saccharification for the production of monosugars. This study characterizes a novel agarase enzyme from <em>S. degradans</em> 2–40, a marine bacterium renowned for its exceptional polysaccharide-degrading capabilities. We hypothesized that this strain would harbor an efficient and robust agarase with desirable properties for saccharification processes. Following isolation and purification, the agarase underwent biochemical analysis, revealing optimal activity at moderate temperatures and a broad pH range. Furthermore, the fusion of Aga50D with NABH enhanced the catalytic efficiency from 1.873 ± 0.22 (mg/mL)⁻¹s⁻¹ to 4.826 ± 0.19 (mg/mL)⁻¹s⁻¹. In contrast to chemical hydrolysis, enzymatic treatment using agarase offers a more selective, eco-friendly, and high-yield alternative, minimizing by-product formation and preserving functional sugar moieties. The enzyme's ability to produce neoagarobiose (NA2) as its primary product, without any intermediates, makes it an attractive biocatalyst for the production of bioactive oligosaccharides. This study highlights the potential of <em>S. degradans</em> 2–40 as a valuable source of enzymes for industrial biotechnology applications, particularly in the sustainable conversion of marine biomass into high-value products.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110733"},"PeriodicalIF":3.7,"publicationDate":"2025-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144826545","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":"NADPH regeneration for efficient biosynthesis of indigo by flavin-containing monooxygenase and formate dehydrogenase","authors":"Yingying Zhu ,&nbsp;Dawei Ni ,&nbsp;Zeyu Li ,&nbsp;Zhebin Hao ,&nbsp;Liang Wang ,&nbsp;Wanmeng Mu","doi":"10.1016/j.enzmictec.2025.110731","DOIUrl":"10.1016/j.enzmictec.2025.110731","url":null,"abstract":"<div><div>Indigo is an important blue pigment widely used in textile, food, and medicine industries. Biological production of indigo attracts increasing attention recently. Cell factory production of indigo encounters the problem of the toxicity of the precursor indole. Enzymatic production is the alternative biological approach, however, NADPH regeneration should be solved. In this study, flavin-containing monooxygenase from <em>Methylophaga aminisulfidivorans</em> was used for enzymatic production of indigo from indole and formate dehydrogenase from <em>Pseudomonas</em> sp. 101 was co-expressed for NADPH regeneration. Indigo production was enhanced by combination of molecular modification, promoter engineering, and translation initiation region engineering. Finally, 0.183 g/L of indigo was produced from 0.5 g/L of indole and 0.5 mM of sodium formate, with the conversion ratio of 32.5 %. This study demonstrates a feasible and effective strategy for enzymatic production of indigo.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"Article 110731"},"PeriodicalIF":3.7,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144780037","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}