Enzyme and Microbial Technology最新文献_第6页

Revolutionizing recombinant protein production in prokaryotic platforms – Methodologies and advances 原核平台重组蛋白生产的革命性变革-方法和进展

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-02-01 Epub Date: 2025-11-05 DOI: 10.1016/j.enzmictec.2025.110778

Shrinidhi Bhat, Senthamizh R, Mayur Mahindra Kedare, Sanjukta Patra

{"title":"Revolutionizing recombinant protein production in prokaryotic platforms – Methodologies and advances","authors":"Shrinidhi Bhat, Senthamizh R, Mayur Mahindra Kedare, Sanjukta Patra","doi":"10.1016/j.enzmictec.2025.110778","DOIUrl":"10.1016/j.enzmictec.2025.110778","url":null,"abstract":"<div><div>Recombinant protein production in prokaryotic systems remains a major topic in biotechnology because of their rapid growth, cost-effectiveness, and ease of genetic manipulation. However, the production of functionally active proteins still faces significant challenges due to folding failures, insolubility, and the lack of the capability of most prokaryotes for complex post-translational processing. This review dwells into both traditional and emerging strategies for optimizing recombinant protein expression in various prokaryotic systems. It also highlights recent advances in genetic engineering and synthetic biology for expanding the toolkit available for protein production, which include refined expression vectors, engineered hosts with improved folding capabilities, and high-throughput screening platforms. Additionally, it provides a thorough discussion of how to optimize heterologous expression using fusion tag approaches, codon bias elimination, promoter and ribosome binding site (RBS) engineering, and chaperone-assisted folding. This review explores diverse prokaryotic expression systems that offer unique advantages for heterologous expression that extend far beyond the limitations of traditional hosts. Additionally, this review also emphasizes the need for the selection of the right expression system and optimizing conditions to fulfill the increasing demands for recombinant protein production in various fields.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"193 ","pages":"Article 110778"},"PeriodicalIF":3.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463925","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}

引用次数: 0

Non-native genetic configuration of Gluconobacter oxydans dehydrogenases drives 2-keto-L-gulonic acid production in recombinant Escherichia coli 脱氧葡萄糖杆菌脱氢酶的非天然遗传结构驱动重组大肠杆菌生产2-酮- l -谷醛酸。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-02-01 Epub Date: 2025-11-07 DOI: 10.1016/j.enzmictec.2025.110776

Saloni Samant , Prashant S. Kharkar , Shreerang V. Joshi , Shamlan M.S. Reshamwala

{"title":"Non-native genetic configuration of Gluconobacter oxydans dehydrogenases drives 2-keto-L-gulonic acid production in recombinant Escherichia coli","authors":"Saloni Samant , Prashant S. Kharkar , Shreerang V. Joshi , Shamlan M.S. Reshamwala","doi":"10.1016/j.enzmictec.2025.110776","DOIUrl":"10.1016/j.enzmictec.2025.110776","url":null,"abstract":"<div><div>Commercial production of 2-keto-L-gulonic acid (2-KGA), a crucial precursor in the synthesis of vitamin C, is carried out using the two-step fermentation process. This process uses a dual-strain system in the second step (conversion of L-sorbose to 2-KGA), necessitating complex process control. In this study, we report development of a recombinant <em>Escherichia coli</em> strain capable of producing 2-KGA from L-sorbose. L-Sorbose dehydrogenase (<em>sdh</em>) and L-sorbosone dehydrogenase (<em>sndh</em>) genes from <em>Gluconobacter oxydans</em> were expressed in <em>E. coli</em> as a synthetic operon in three different configurations under the control of the strong inducible T7 promoter. High-density whole-cell biotransformation was carried out to produce 2-KGA from L-sorbose, resulting in a yield of 0.69 g/g when the two genes were arranged in the non-native <em>sdh-sndh</em> configuration. Our design eliminates the need for multi-organism co-culture, intricate redox balancing, or nutrient-enriched media, representing a robust and scalable alternative to conventional production. This work demonstrates the feasibility of employing <em>E. coli</em> as a chassis strain for vitamin C precursor biosynthesis and offers a modular operon framework adaptable to other dehydrogenase-driven bioconversions. This approach enables efficient, scalable production of the vitamin C precursor in a genetically tractable host.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"193 ","pages":"Article 110776"},"PeriodicalIF":3.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145494911","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}

引用次数: 0

An H1N1 virus biosensor based on enzyme activity-gated PER-CRISPR/Cas12a cascade signal amplification 基于酶活性门控PER-CRISPR/Cas12a级联信号扩增的H1N1病毒生物传感器

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-10-09 DOI: 10.1016/j.enzmictec.2025.110759

Meiyan Xin , Jitong Liu , Hongyu Zhou , Shengjun Bu , Zhuo Hao , He Sun , Jie Lu , Xiangru Feng , Xue Jiang , Qingshuang Wang , Jiayu Wan

{"title":"An H1N1 virus biosensor based on enzyme activity-gated PER-CRISPR/Cas12a cascade signal amplification","authors":"Meiyan Xin , Jitong Liu , Hongyu Zhou , Shengjun Bu , Zhuo Hao , He Sun , Jie Lu , Xiangru Feng , Xue Jiang , Qingshuang Wang , Jiayu Wan","doi":"10.1016/j.enzmictec.2025.110759","DOIUrl":"10.1016/j.enzmictec.2025.110759","url":null,"abstract":"<div><div>The rapid and accurate detection of the H1N1 influenza virus is a key link in epidemic prevention and control. This study innovatively constructed a cascade signal amplification biosensor based on DNA polymerase activity regulation, aiming to achieve ultra-sensitive and highly specific detection of viral nucleic acids. This biosensor has the following significant advantages: (i) Molecular lock-key regulation mechanism: A functional DNA inhibitor is designed to form a complex with Taq DNA polymerase, and the target H1N1 RNA is specifically recognized to release enzyme activity inhibition, converting the target presence information into a PER reaction initiation signal. (ii) Cascade signal amplification system: The single-stranded DNA generated by PER activates Cas12a trans-cleavage activity, achieving a three-level signal amplification of enzyme activity activation → nucleic acid synthesis → CRISPR cleavage. The biosensor exhibits a linear detection range between 1 pM and 1 μM, with a detection limit of 25 fM. Moreover, the platform showed high versatility and could be readily adapted for the detection of other pathogens such as SARS-CoV-2 by simply modifying the nucleic acid sequences of the inhibitor and activator. This study not only provides a new tool for the screening of H1N1 influenza virus, but also offers a novel strategy for the development of next-generation molecular detection technologies suitable for point-of-care diagnostics, indicating considerable application potential.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110759"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145266795","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}

引用次数: 0

Enhancing isoprimeverose-producing enzyme in genetically engineered Aspergillus oryzae through impeller shape and pH control for the large-scale fermentation 通过叶轮形状和pH控制来提高转基因米曲霉大规模发酵产异戊糖酶的活性

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-10-05 DOI: 10.1016/j.enzmictec.2025.110758

Fahmi Baihaqqi , Satoshi Wakai , Filemon Jalu Nusantara Putra , Tomohiro Suzuki , Yoshiro Ikeya , Prihardi Kahar , Yutaro Mori , Akihiko Kondo , Chiaki Ogino

{"title":"Enhancing isoprimeverose-producing enzyme in genetically engineered Aspergillus oryzae through impeller shape and pH control for the large-scale fermentation","authors":"Fahmi Baihaqqi , Satoshi Wakai , Filemon Jalu Nusantara Putra , Tomohiro Suzuki , Yoshiro Ikeya , Prihardi Kahar , Yutaro Mori , Akihiko Kondo , Chiaki Ogino","doi":"10.1016/j.enzmictec.2025.110758","DOIUrl":"10.1016/j.enzmictec.2025.110758","url":null,"abstract":"<div><div><em>Aspergillus oryzae</em> is a filamentous fungus that serves as a source of polysaccharide-degrading enzymes. We constructed two self-cloning forms of <em>A. oryzae</em> that specifically express isoprimeverose oligoxyloglucanase (IpeA) and endoglucanase. These enzymes are essential for the enzymatic release of the rare sugar isoprimeverose (α-D-xylopyranosyl-(1→6)-D-glucopyranose) from tamarind xyloglucan. Of the two, IpeA is a key enzyme that is currently unavailable commercially. The primary objective of this study was to enhance IpeA production through fermentation engineering. Various fermentation parameters were optimized. These include a source of carbon from either glucose or maltose, pH control, and impeller shapes using either Disk Turbine (DT) or MAXBLEND® (MB) impellers that were tested using a 5-L stirred-tank bioreactor. Cultivation in maltose-supplemented medium resulted in a reduction in carbon catabolite repression compared with that of glucose. Batch fermentation at acidic pH (4.5) using the MB impeller yielded a significantly higher level of biomass and enzyme production with a dry mycelial weight (DMW) yield of ∼19 g/L and IpeA activity of ∼68 U/mL, which compares favorably with ∼14 g/L of DMW and 43 U/mL of IpeA activity under identical pH conditions when using a DT impeller. The findings of this study offer valuable insight for both large-scale and biological production of <em>A. oryzae</em>.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110758"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145266796","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}

引用次数: 0

Expression and biochemical characterization of wild-type and mutant β-amylases from Priestia flexa in Komagataella phaffii 野生型和突变型柔曲Priestia β-淀粉酶在法菲Komagataella phaffii中的表达及生化特性研究。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-10-11 DOI: 10.1016/j.enzmictec.2025.110761

Tatiana Leonidovna Gordeeva, Artur Aleksandrovich Tkachenko, Larisa Nikolaevna Borshchevskaya, Natalia Vladimirovna Bulushova, Egor Sergeevich Bobrov, Aleksandr Sergeevich Fedorov, Ekaterina Pavlovna Sakharova, Olga Evgenevna Melkina, Sergey Pavlovich Sineoky

{"title":"Expression and biochemical characterization of wild-type and mutant β-amylases from Priestia flexa in Komagataella phaffii","authors":"Tatiana Leonidovna Gordeeva, Artur Aleksandrovich Tkachenko, Larisa Nikolaevna Borshchevskaya, Natalia Vladimirovna Bulushova, Egor Sergeevich Bobrov, Aleksandr Sergeevich Fedorov, Ekaterina Pavlovna Sakharova, Olga Evgenevna Melkina, Sergey Pavlovich Sineoky","doi":"10.1016/j.enzmictec.2025.110761","DOIUrl":"10.1016/j.enzmictec.2025.110761","url":null,"abstract":"<div><div>The β-amylases AmyPf1 and AmyPf2 from two distinct <em>Priestia flexa</em> strains were successfully expressed for the first time in <em>Komagataella phaffii</em>. The main biochemical characteristics of the recombinant enzymes were determined. β-Amylases exhibited similar optimal temperature (55 °C) and pH (pH 8.0), as well as high stability at 55 °C and across a broad pH range (4.0–10.0). The thermal inactivation half-lives at 55 °C for both recombinant enzymes were approximately 4 h. The AmyPf2 demonstrated superior specific activity (3836 U•mg<sup>−1</sup>) and catalytic constant (6367 s<sup>−1</sup>) compared to AmyPf1 (3411 U•mg<sup>−1</sup> and 4224 s<sup>−1</sup>, respectively). Comparison of the amino acid sequences of the two β-amylases revealed four residue differences. Site-directed mutagenesis introducing AmyPf2-specific substitutions into the AmyPf1 sequence showed that the V180A mutation significantly enhanced specific activity (4009 U•mg<sup>−1</sup>) and catalytic constant (6656 s<sup>−1</sup>) of the resulting r-V180A variant. In fed-batch fermentation, the activity of r-V180A reached 5130 U•mL<sup>−1</sup>. Hydrolysis of corn starch using r-V180A in combination with pullulanase (a starch-debranching enzyme) yielded 87.56 % maltose, indicating that a hydrolysate with high maltose content was obtained. These results highlight the potential of the recombinant r-V180A enzyme for industrial maltose production and underscore the utility of the <em>K. phaffii</em> expression system for efficient production of <em>P. flexa</em> β-amylases.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110761"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145291600","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}

引用次数: 0

Anti-biofilm efficiency and substrate specificity of recombinantly produced human paraoxonases 重组人对氧酶的抗生物膜效率和底物特异性。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-10-08 DOI: 10.1016/j.enzmictec.2025.110760

Manik Goel, Priyamedha Yadav, Fauzia Parween, Sudisha Mukherjee, Rinkoo Devi Gupta

{"title":"Anti-biofilm efficiency and substrate specificity of recombinantly produced human paraoxonases","authors":"Manik Goel, Priyamedha Yadav, Fauzia Parween, Sudisha Mukherjee, Rinkoo Devi Gupta","doi":"10.1016/j.enzmictec.2025.110760","DOIUrl":"10.1016/j.enzmictec.2025.110760","url":null,"abstract":"<div><div>The human paraoxonase (PON) gene family, comprising PON1, PON2, and PON3, has evolved through parallel evolutionary trajectories leading to distinct substrate selectivity, thereby enabling these enzymes to hydrolyze a range of substrates such as lactones, arylesters, and organophosphates. This study explores the evolutionary divergence of PON family members and their varied anti-biofilm activities through <em>in silico</em> molecular docking and <em>in vitro</em> analysis. The huPON1, huPON2, and huPON3 genes were cloned and expressed in the <em>E. coli</em> expression system. These overexpressed proteins formed inclusion bodies, which were further subjected to <em>in vitro</em> refolding and biochemical characterization. The enzymatic studies revealed that huPON1 has unique paraoxonase activity and high arylesterase activity, while huPON2 demonstrated the highest lactonase activity, followed by huPON3 and huPON1. A comparative assessment of the anti-biofilm potential of recombinant huPONs was conducted against <em>Mycobacterium smegmatis</em>, which revealed that all three paraoxonases inhibited biofilm formation; however, huPON2 displayed the highest anti-biofilm activity, followed by huPON3 and huPON1. Additionally, all the huPONs showed a synergistic effect with the conventional TB drug, Rifampicin, in enhancing biofilm disruption. These findings contribute valuable insights into the application of huPONs as a therapeutic tool against drug-resistant bacterial infections.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110760"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145279236","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}

引用次数: 0

Mechanism of tyrosine 69 in the flavin domain modulating electron transfer efficiency in BVU5 azoreductase 黄素区域酪氨酸69调节BVU5偶氮还原酶电子转移效率的机制

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-09-25 DOI: 10.1016/j.enzmictec.2025.110757

Xuehui Xie , Dongyang Li , Hangmi Zheng , Ziyi Wu , Jiajie Liu , Wanting Li , Songwei Chen , Yiwen Zhang , Daijie Tang , Na Liu , Qingyun Zhang , Ye Chen

{"title":"Mechanism of tyrosine 69 in the flavin domain modulating electron transfer efficiency in BVU5 azoreductase","authors":"Xuehui Xie , Dongyang Li , Hangmi Zheng , Ziyi Wu , Jiajie Liu , Wanting Li , Songwei Chen , Yiwen Zhang , Daijie Tang , Na Liu , Qingyun Zhang , Ye Chen","doi":"10.1016/j.enzmictec.2025.110757","DOIUrl":"10.1016/j.enzmictec.2025.110757","url":null,"abstract":"<div><div>The azoreductase BVU5 is a flavoprotein dependent on NAD(P)H/FMN-mediated electron transfer; However, the functional mechanism of key residues within its flavin domain remains unclear. This study identified the conserved residue Tyr<sup>69</sup> through bioinformatics analysis and constructed Y69F and Y69C mutants. Enzymatic assays demonstrated that mutants exhibited significantly lower decolorization rates than the BVU5 enzyme across 12 dye molecules, including azo, anthraquinone, and triphenylmethane dyes. For example, when decolorizing Reactive Black 5 (RB5) for 2 h: Y69F mutant achieved 60 %-65 % decolorization, Y69C mutant reached 55 %-60 % decolorization, both markedly lower than the 80 %-85 % efficiency of BVU5.The decolorization hierarchy remained azo dyes > triphenylmethane > anthraquinone dyes. Molecular docking revealed that mutations reconfigured FMN-binding patterns. Although Y69F enlarged the substrate-binding pocket, it failed to enhance the degradation efficiency of the bulky dye Chlorazol Black E. This critical contradiction indicates that substrate binding is not the limiting factor. Combined with evidence such as the lighter color of the mutant enzyme solutions, increased A280/A450 ratios, and enzyme dosage experiments, this study confirms that Tyr<sup>69</sup> is a key residue that sustains electron transfer efficiency by maintaining the FMN-binding conformation, thereby determining the decolorization performance. Consequently, electron transfer efficiency, rather than substrate binding, is the primary mechanism influencing the catalytic function of BVU5.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110757"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155168","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}

引用次数: 0

Homologous expression and characterization of Coniochaeta ligniaria glycoside hydrolase family 115 α-glucuronidase 木质素螺毛菌糖苷水解酶家族115 α-葡萄糖醛酸酶的同源表达与鉴定

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-09-09 DOI: 10.1016/j.enzmictec.2025.110750

Michael J. Bowman, Nancy N. Nichols

{"title":"Homologous expression and characterization of Coniochaeta ligniaria glycoside hydrolase family 115 α-glucuronidase","authors":"Michael J. Bowman, 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><sub><em>m</em></sub> 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":"2026-01-01","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}

引用次数: 0

De Novo enzyme design of a minimal and soluble carbonic anhydrase from Mesorhizobium loti assisted by molecular dynamics simulation 在分子动力学模拟的辅助下，从loti中分离出一种最小的可溶碳酸酐酶

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-09-12 DOI: 10.1016/j.enzmictec.2025.110751

Jiun-Jang Juo, I.-Son Ng

{"title":"De Novo enzyme design of a minimal and soluble carbonic anhydrase from Mesorhizobium loti assisted by molecular dynamics simulation","authors":"Jiun-Jang Juo, I.-Son Ng","doi":"10.1016/j.enzmictec.2025.110751","DOIUrl":"10.1016/j.enzmictec.2025.110751","url":null,"abstract":"<div><div>Carbonic anhydrases (CAs) are critical biocatalysts in the carbon capture and utilization due to their remarkable efficiency in converting carbon dioxide into bicarbonate. Among all, the CA from <em>Mesorhizobium loti</em> (MlCA) exhibits the highest catalytic activity. However, the poor expression and low solubility in <em>Escherichia coli</em> significantly restricts its application. To overcome the challenges, we employed a protein minimization strategy to improve both enzymatic expression and solubility. Enhanced expressions were observed in genetic constructs with short N-terminal tags which MlCA expression is strongly affected by mRNA secondary structure near the start codon. For solubility issue, a <em>de novo</em> protein design workflow guided by molecular dynamics simulations was developed. The process consists of four stages: (1) size-constrained <em>de novo</em> design considered using AlphaFold and RFdiffusion, (2) sequence recovery replying on solubleMPNN and ESMfold, (3) <em>In silico</em> screening by FoldX, SASA and molecular dynamics evaluation, and (4) experimental validation. This enabled the generation of compact, stable, and catalytically active MlCA variants with 28 % reduction in protein size among 500 candidates. Finally, the most promising design, dM22, of minimal CA showed improved solubility from 16.1 % to 61.2 % in B7G, a GroELS integrated BL21(DE3) strain.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110751"},"PeriodicalIF":3.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145057156","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}

引用次数: 0

Innovative nanosensing methods for lysozyme identification 溶菌酶鉴定的创新纳米传感方法

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2026-01-01 Epub Date: 2025-09-02 DOI: 10.1016/j.enzmictec.2025.110745

Mohammad Darvishi , Seyyed Mohammad Mousavinia , Reza Soleimany , Mohammad Mahdi Heidari , Mohsen Mohammadi , Samad Rastmanesh , Ahmad Mobed

{"title":"Innovative nanosensing methods for lysozyme identification","authors":"Mohammad Darvishi , Seyyed Mohammad Mousavinia , Reza Soleimany , Mohammad Mahdi Heidari , Mohsen Mohammadi , Samad Rastmanesh , 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":"2026-01-01","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}

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