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Improvement of β-Xylosidase and Endoxylanase Activities in Talaromyces amestolkiae by Genetic Manipulation of the Transcriptional Activator XlnR 转录激活子XlnR的遗传修饰对阿梅塔芳酵母β-木糖苷酶和内木聚糖酶活性的影响
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-26 DOI: 10.1111/1751-7915.70166
Ana Pozo-Rodríguez, Miguel Ángel Peñalva, Jorge Barriuso, Eduardo A. Espeso, María Jesús Martínez
{"title":"Improvement of β-Xylosidase and Endoxylanase Activities in Talaromyces amestolkiae by Genetic Manipulation of the Transcriptional Activator XlnR","authors":"Ana Pozo-Rodríguez,&nbsp;Miguel Ángel Peñalva,&nbsp;Jorge Barriuso,&nbsp;Eduardo A. Espeso,&nbsp;María Jesús Martínez","doi":"10.1111/1751-7915.70166","DOIUrl":"https://doi.org/10.1111/1751-7915.70166","url":null,"abstract":"<p>The ascomycete <i>Talaromyces amestolkiae</i> is a promising source of glycosyl hydrolases for hemicellulose degradation, as it contains a considerably higher number of genes encoding these enzymes than other fungi exploited for plant biomass valorisation. The development of genetic engineering tools could further improve its biotechnological potential. We report here a transformation system for <i>T. amestolkiae</i> based on pyrimidine auxotrophy complementation, which was used to successfully introduce both integrative and autonomously replicating plasmids. Then, we applied this tool to force the expression of the transcriptional activator XlnR, generating an engineered strain with enhanced β-xylosidase (1.4-fold) and endoxylanase (2.0-fold) activities compared to the wild-type cultured on xylan. Markedly larger improvements were obtained after introducing Ala788Val or Val785Phe substitutions in XlnR, achieving 3.3-fold and 3.9-fold increases in β-xylosidase and endoxylanase activities, respectively, in the case of XlnR<sup>V785F</sup>. This recombinant strain also displays a partial deregulation of the hemicellulolytic system when cultivated on glucose and glycerol (a low-cost and renewable substrate), yielding notably higher production of β-xylosidases (16.9-fold and 13.8-fold) and endoxylanases (31.9-fold and 22.7-fold) than the wild-type. Increased efficiencies of XlnR<sup>V785F</sup> enzymatic crudes in xylan saccharifications showed the potential of XlnR engineering to develop robust <i>T. amestolkiae</i> strains for the valorisation of hemicellulosic residues.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70166","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144135694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Biofertilizer Industry and Research Developments in China: A Mini-Review 中国生物肥料产业与研究进展综述
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-24 DOI: 10.1111/1751-7915.70163
Xinli Sun, Zhihui Xu, Nan Zhang, Youzhi Miao, Chao Zhang, Xiaoli Ma, Qirong Shen, Ruifu Zhang
{"title":"Biofertilizer Industry and Research Developments in China: A Mini-Review","authors":"Xinli Sun,&nbsp;Zhihui Xu,&nbsp;Nan Zhang,&nbsp;Youzhi Miao,&nbsp;Chao Zhang,&nbsp;Xiaoli Ma,&nbsp;Qirong Shen,&nbsp;Ruifu Zhang","doi":"10.1111/1751-7915.70163","DOIUrl":"https://doi.org/10.1111/1751-7915.70163","url":null,"abstract":"<p>Reliance on chemical fertilizers has significantly boosted food production in China, but it has also led to soil degradation, environmental pollution, and greenhouse gas emissions. To address these pressing issues, the Chinese government has launched various initiatives to reduce chemical fertilizer consumption and promote biofertilizers as effective alternatives to enhance soil fertility and mitigate environmental pollution. Biofertilizers promote crop growth by providing or activating essential nutrients, suppressing plant pathogens, improving soil health, and increasing resilience to abiotic stresses. The growing adoption of biofertilizers in China is reflected in the registration of more than 10,000 products, an annual production exceeding 35 million tons, and a market value of over US$5.5 billion, indicating a significant shift towards sustainable agricultural practices. Despite this progress, challenges such as the dominance of nitrogen fertilizers, inconsistent product performance, and the need for cultivar-specific microbial inoculants remain. Foundational research on the microbial genera utilised in biofertilizers, including nitrogen-fixing genera <i>Rhizobium</i>, <i>Paenibacillus</i>, and <i>Pseudomonas</i>, the widely used genus, <i>Bacillus</i> and <i>Trichoderma</i>, as well as multipurpose synthetic communities, is essential for overcoming these obstacles and enhancing the efficacy of biofertilizers. This review delves into the historical development of the biofertilizer industry and recent advancements in fundamental research on biofertilizers in China, highlighting the essential role of biofertilizers in promoting green agricultural development.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70163","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144125857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protein Biosynthesis and Carbon Catabolite Repression Are Transcriptionally Upregulated in Saccharomyces cerevisiae by Extracellular Fractions From Several Wine Yeast Species 几种酒酵母菌胞外组分转录上调酿酒酵母菌的蛋白质生物合成和碳分解代谢抑制
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-23 DOI: 10.1111/1751-7915.70168
Miguel Mejias-Ortiz, Pilar Morales, Guillermo Juárez, Ramon Gonzalez
{"title":"Protein Biosynthesis and Carbon Catabolite Repression Are Transcriptionally Upregulated in Saccharomyces cerevisiae by Extracellular Fractions From Several Wine Yeast Species","authors":"Miguel Mejias-Ortiz,&nbsp;Pilar Morales,&nbsp;Guillermo Juárez,&nbsp;Ramon Gonzalez","doi":"10.1111/1751-7915.70168","DOIUrl":"https://doi.org/10.1111/1751-7915.70168","url":null,"abstract":"<p>Non-<i>Saccharomyces</i> yeast species are increasingly used in winemaking in combination with <i>Saccharomyces cerevisiae</i> to modulate sensory attributes or as processing aids. Consequently, there is academic and practical interest in understanding how different yeast species interact with each other in grape must. Although interactions will depend on the metabolic capabilities of the strains involved, there are other possible interaction mechanisms between wine yeasts. In this work we used extracellular vesicle (EV)-enriched fractions from different non-<i>Saccharomyces</i> species to challenge <i>S. cerevisiae</i> inoculated in synthetic grape must. The results show that the previously described response to EVs of <i>Metschnikowia pulcherrima</i> was not an isolated phenomenon, but that <i>S. cerevisiae</i> responds in a general way to EVs of other yeast species. Meta-analysis of the results points to protein biosynthesis and carbon catabolite repression as general targets; both being stimulated by the interaction, beyond the acclimatisation to the synthetic juice experienced by the control cells. The intensity of the response showed differences between the four species; while the transcriptional response to <i>M. pulcherrima</i> EVs clearly diverges from that to EVs of the other yeast species, which show greater similarity to each other.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70168","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bacterial Species in Engineered Living Materials: Strategies and Future Directions 工程生物材料中的细菌种类:策略和未来方向
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-23 DOI: 10.1111/1751-7915.70164
Hu Wang, Chunzhong Li, Yanmin Wang, Huanming Zhang
{"title":"Bacterial Species in Engineered Living Materials: Strategies and Future Directions","authors":"Hu Wang,&nbsp;Chunzhong Li,&nbsp;Yanmin Wang,&nbsp;Huanming Zhang","doi":"10.1111/1751-7915.70164","DOIUrl":"https://doi.org/10.1111/1751-7915.70164","url":null,"abstract":"<p>In recent years, there has been a notable increase interest in engineered living materials (ELMs) owing to their considerable potential. One key area of research within this field is the utilisation of various species of bacteria to create innovative living materials. In order to accelerate the advancement of bacterial-based living materials, a systematic summary of bacterial species and their design strategies is essential. Yet, up to this point, no applicable reviews have been documented. This review offers a concise introduction to living materials derived from bacteria, delves into the strategies and applications of each bacterial species in this realm, and provides perspectives and future outlooks in this field.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70164","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery 腐臭假单胞菌KT2440利用代谢工程和原位产物回收从阿魏酸中生产香兰素
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-23 DOI: 10.1111/1751-7915.70152
Ilona A. Ruhl, Sean P. Woodworth, Stefan J. Haugen, Hannah M. Alt, Gregg T. Beckham, Christopher W. Johnson
{"title":"Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery","authors":"Ilona A. Ruhl,&nbsp;Sean P. Woodworth,&nbsp;Stefan J. Haugen,&nbsp;Hannah M. Alt,&nbsp;Gregg T. Beckham,&nbsp;Christopher W. Johnson","doi":"10.1111/1751-7915.70152","DOIUrl":"https://doi.org/10.1111/1751-7915.70152","url":null,"abstract":"<p>Vanillin is the most in-demand flavouring compound in the world and because vanillin extracted from vanilla pods cannot meet the global demand, most vanillin on the market today is chemically synthesised. Increasing demands by consumers for natural ingredients have inspired efforts to develop vanillin derived from microbial sources. These efforts have been challenged by low titers, likely caused by the toxicity of vanillin to most microbial biocatalysts. In this study, we engineered a <i>Pseudomonas putida</i> KT2440-derived strain that accumulated vanillin from ferulic acid to 0.64 g/L. To increase the overall titre, we applied a hydrophobic polystyrene-based resin to vanillin-accumulating cultures, which enabled an increase in total vanillin recovery to an apparent titre of 3.35 g/L. This study demonstrates that <i>P. putida</i> can accumulate vanillin from ferulic acid to higher titers when vanillin is removed from the cultivation medium, mitigating its toxicity.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70152","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144125892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Toxoplasma gondii C2 Domain Protein Deletion Mutant as a Promising Vaccine Against Toxoplasmosis in Mice 刚地弓形虫C2结构域蛋白缺失突变体作为抗小鼠弓形虫病疫苗的前景
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-23 DOI: 10.1111/1751-7915.70143
Yifan Luo, Mingfeng He, Shengqiang Yang, Jiahui Qian, Zhengming He, Jiayin Xu, Liyu Guo, Siyu Xiao, Rui Fang
{"title":"Toxoplasma gondii C2 Domain Protein Deletion Mutant as a Promising Vaccine Against Toxoplasmosis in Mice","authors":"Yifan Luo,&nbsp;Mingfeng He,&nbsp;Shengqiang Yang,&nbsp;Jiahui Qian,&nbsp;Zhengming He,&nbsp;Jiayin Xu,&nbsp;Liyu Guo,&nbsp;Siyu Xiao,&nbsp;Rui Fang","doi":"10.1111/1751-7915.70143","DOIUrl":"https://doi.org/10.1111/1751-7915.70143","url":null,"abstract":"<p><i>Toxoplasma gondii</i> (<i>T. gondii</i>), a parasitic protozoan capable of infecting nearly all warm-blooded animals, causes significant economic losses in livestock and poses a significant threat to both animal and public health. Despite its impact, no ideal vaccine is currently available to prevent toxoplasmosis. Vesicular transport plays a crucial role in the life cycle of <i>T. gondii</i>, and proteins involved in this process – such as those containing C2 domains – may serve as novel targets for the development of live attenuated vaccines. In this study, we evaluated the feasibility of a C2 domain-containing protein (TGME49_203240) as a live attenuated vaccine candidate. Our findings suggest that TGME49_203240 may be involved in vesicular transport and that it is essential for <i>T. gondii</i> growth. Deletion of TGME49_203240 reduced parasite virulence and impaired tissue cyst formation in mice. Moreover, mice vaccinated with ME49<i>Δ203240</i> were protected against the lethal challenge of the tachyzoites of <i>T. gondii</i> I, II, III strains and cysts of II strain. In addition, the ME49<i>Δ203240</i> strain elicited robust immune responses, including the production of high levels of specific IgG antibodies and key cytokines (IFN-γ, TNF-α and IL-12). These findings highlight TGME49_203240 as a promising target for the development of a live attenuated vaccine against <i>T. gondii</i>.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70143","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Reconstitution of Methionine Cycle With ATP Regeneration for Whole-Cell Catalysis of Creatine Production in Engineered Escherichia coli ATP再生重组蛋氨酸循环在工程大肠杆菌全细胞催化肌酸生产中的应用
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-21 DOI: 10.1111/1751-7915.70145
Yuhua Sheng, Yaokang Wu, Linpei Zhang, Xueqin Lv, Jianghua Li, Long Liu, Guocheng Du, Jian Chen, Yanfeng Liu
{"title":"Reconstitution of Methionine Cycle With ATP Regeneration for Whole-Cell Catalysis of Creatine Production in Engineered Escherichia coli","authors":"Yuhua Sheng,&nbsp;Yaokang Wu,&nbsp;Linpei Zhang,&nbsp;Xueqin Lv,&nbsp;Jianghua Li,&nbsp;Long Liu,&nbsp;Guocheng Du,&nbsp;Jian Chen,&nbsp;Yanfeng Liu","doi":"10.1111/1751-7915.70145","DOIUrl":"https://doi.org/10.1111/1751-7915.70145","url":null,"abstract":"<p>Creatine (CR) is a naturally occurring amino acid derivative that plays a key role in cellular energy homeostasis, which has wide-ranging applications in food and medicine. Currently, the lack of green and sustainable CR biomanufacturing methods has led to reliance on chemical methods for industrial CR synthesis. This study presents a biological approach to synthesising CR using whole-cell catalysis by engineered <i>Escherichia coli</i>. First, through screening of critical enzymes from different sources and dual-enzyme co-expression strategies, arginine: glycine amidinotransferase (AGAT) from <i>Amycolatopsis kentuckyensis</i> and guanidinoacetate N-methyltransferase (GAMT) from <i>Mus caroli</i> were introduced to construct the CR biosynthesis pathway, yielding 0.83 g/L CR production. Then, the expression level of GAMT, the critical rate-limiting enzyme, was optimised by screening the ribosome binding site and N-terminal coding sequences, resulting in a 92% enhancement of CR production, reaching 1.59 g/L. Next, the endogenous ornithine and methionine cycles were further engineered to boost the synthesis of the precursor guanidinoacetate (GAA) and methyl donor S-adenosylmethionine (SAM), leading to a 68% increase in CR production, reaching 2.67 g/L. Finally, considering adenosine triphosphate (ATP) is required as a cofactor for SAM biosynthesis, we integrated the reconstitution methionine cycle with a polyphosphate kinase-based ATP regeneration system, achieving a CR titre of 5.27 g/L with a productivity of 0.22 g/L/h, and the molar conversion of substrate arginine was 71 mol% over 24 h following the engineering process. This study is the first report achieving whole-cell catalysis of CR production in engineered <i>E. coli</i> with a dual enzyme cascade using arginine as substrate, providing a new platform for CR production and insights into the biosynthesis of high-value metabolites that rely on ATP consumption.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70145","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144100874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Pseudomonas aeruginosa Performs Chemotaxis to the Neurotransmitters Serotonin, Dopamine, Epinephrine and Norepinephrine 铜绿假单胞菌对神经递质5 -羟色胺、多巴胺、肾上腺素和去甲肾上腺素具有趋化作用
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-19 DOI: 10.1111/1751-7915.70161
Elizabet Monteagudo-Cascales, Andrea Lozano-Montoya, Tino Krell
{"title":"Pseudomonas aeruginosa Performs Chemotaxis to the Neurotransmitters Serotonin, Dopamine, Epinephrine and Norepinephrine","authors":"Elizabet Monteagudo-Cascales,&nbsp;Andrea Lozano-Montoya,&nbsp;Tino Krell","doi":"10.1111/1751-7915.70161","DOIUrl":"https://doi.org/10.1111/1751-7915.70161","url":null,"abstract":"<p>Bacteria use chemotaxis to move to favourable ecological niches. For many pathogenic bacteria, chemotaxis is required for full virulence, particularly for the initiation of host colonisation. There do not appear to be limits to the type of compounds that attract bacteria, and we are just beginning to understand how chemotaxis adapts them to their lifestyles. Quantitative capillary assays for chemotaxis show that <i>P. aeruginosa</i> is strongly attracted to serotonin, dopamine, epinephrine and norepinephrine. Chemotaxis to these compounds is greatly decreased in a mutant lacking the TlpQ chemoreceptor, and complementation of this mutant with a plasmid harbouring the <i>tlpQ</i> gene restores wild-type-like chemotaxis. Microcalorimetric titrations of the TlpQ sensor domain with these four compounds indicate that they bind directly to TlpQ. All four compounds are hormones and neurotransmitters that control a variety of processes and are also important signal molecules involved in the virulence of <i>P. aeruginosa</i>. They modulate motility, biofilm formation, the production of virulence factors and serve as siderophores that chelate iron. Additionally, this is the first report of bacterial chemotaxis to serotonin. This study provides an incentive for research to define the contribution of chemotaxis to these host signalling molecules to the virulence of <i>P. aeruginosa</i>.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70161","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144085142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A Novel Chitin-Based Purification System Using GAL1 Fusion Tags: Enhancing Recombinant Protein Production While Retaining Biological Activity 利用GAL1融合标签的新型几丁质纯化系统:在保持生物活性的同时提高重组蛋白的产量
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-15 DOI: 10.1111/1751-7915.70157
Yao-Kuang Tseng, Yun-Heng Lu, Yun Liu, Zhi-Wei Weng, Yu-Tzu Lin, Chih-Hsuan Tsai, Yueh-Lung Wu, Rong-Nan Huang
{"title":"A Novel Chitin-Based Purification System Using GAL1 Fusion Tags: Enhancing Recombinant Protein Production While Retaining Biological Activity","authors":"Yao-Kuang Tseng,&nbsp;Yun-Heng Lu,&nbsp;Yun Liu,&nbsp;Zhi-Wei Weng,&nbsp;Yu-Tzu Lin,&nbsp;Chih-Hsuan Tsai,&nbsp;Yueh-Lung Wu,&nbsp;Rong-Nan Huang","doi":"10.1111/1751-7915.70157","DOIUrl":"https://doi.org/10.1111/1751-7915.70157","url":null,"abstract":"<p>Efficient and economical purification methods are crucial for the commercial production of recombinant proteins with biomedical applications. In this study, we developed an affinity chromatography system that leverages the polysaccharide-binding properties of galectin-1 (GAL1) as a protein tag. The known GAL1-binding material, chitin, was used as the purification matrix. Melittin (MELT), a bee venom peptide known for its antimicrobial and anti-inflammatory properties with commercial potential, was chosen to validate this system. The GAL1–MELT fusion protein was expressed in <i>Escherichia coli</i> (<i>E. coli</i>) and successfully purified using a chitin-based matrix with sodium dodecyl sulfate (SDS) as a removable eluant. This method demonstrated higher purification efficiency compared to the His-tag/Ni-NTA approach, indicating that the GAL1/chitin system could serve as a superior alternative. The GAL1–MELT fusion protein retained strong antibacterial and anti-inflammatory activities, as well as collagen content modulation effects, confirming that MELT maintained its bioactivity. Apart from that, the GAL1–DsRed fusion protein was used as an additional protein target to evaluate the efficiency of the chitin-based column. Notably, all experiments were conducted without tag cleavage, showing that enzyme treatments for MELT isolation were unnecessary. This study highlights the potential of GAL1–polysaccharide interactions as a cost-effective and highly efficient alternative method for recombinant protein purification.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70157","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143950215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Disarming the Pathogenicity of Methicillin-Resistant Staphylococcus aureus via Osmundacetone-Mediated Inhibition of Sortase A 通过奥蒙丙酮介导的对分类酶A的抑制解除耐甲氧西林金黄色葡萄球菌的致病性
IF 5.7 2区 生物学
Microbial Biotechnology Pub Date : 2025-05-13 DOI: 10.1111/1751-7915.70119
Rong Wang, Chunhui Zhao, Dongbin Guo, Yueying Wang, Luanbiao Sun, Xinyao Liu, Yun Sun, Da Liu, Jiyu Guan, Li Wang, Bingmei Wang
{"title":"Disarming the Pathogenicity of Methicillin-Resistant Staphylococcus aureus via Osmundacetone-Mediated Inhibition of Sortase A","authors":"Rong Wang,&nbsp;Chunhui Zhao,&nbsp;Dongbin Guo,&nbsp;Yueying Wang,&nbsp;Luanbiao Sun,&nbsp;Xinyao Liu,&nbsp;Yun Sun,&nbsp;Da Liu,&nbsp;Jiyu Guan,&nbsp;Li Wang,&nbsp;Bingmei Wang","doi":"10.1111/1751-7915.70119","DOIUrl":"https://doi.org/10.1111/1751-7915.70119","url":null,"abstract":"<p>Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) is a major global health threat due to its resistance to multiple antibiotics, making conventional treatments ineffective. The rise in antibiotic resistance highlights the urgent need for new therapies. Sortase A (SrtA), a key virulence factor in <i>Staphylococcus aureus</i> (<i>S. aureus</i>), facilitates bacterial adhesion and infection by anchoring surface proteins to host cells, making it a promising drug target. In this study, we investigated the potential of osmundacetone (OSC), a natural compound from Osmundae Rhizoma, as an SrtA inhibitor. Using fluorescence resonance energy transfer (FRET), OSC was found to inhibit SrtA with an IC<sub>50</sub> of 1.29 μg/mL (7.24 μM). Further in vitro assays confirmed the effectiveness of OSC in inhibiting SrtA-mediated bacterial adhesion, invasion and biofilm formation. Fluorescence quenching and molecular docking pinpointed the binding site of OSC on SrtA. In vivo, OSC improved survival rates in MRSA-infected mice and <i>Galleria mellonella</i> (<i>G. mellonella)</i> while reducing bacterial loads in infected tissues. These results suggest OSC as a promising candidate for anti-MRSA therapies.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 5","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70119","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143938955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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