Biotechnology Letters最新文献

{"title":"A new strategy for the fermentation of Massa Medicata Fermentata by combining multiple strains of fermentation and their fermentation mechanisms.","authors":"Wanqiu Liu, Ying Liu, Jie Zhang, Huangyao Zhu, Siqi Fan, Jingwen Ha, Yuxuan Lu, Yizhu Wang, Wenyuan Liu, Ma Mi, Feng Feng, Jian Xu","doi":"10.1007/s10529-025-03595-3","DOIUrl":"10.1007/s10529-025-03595-3","url":null,"abstract":"<p><p>To address the quality instability of the traditional fermentation process of Massa Medicata Fermentata (LSQ), we designed an innovation strategy for dual-strain co-fermentation LSQ. Rhizopus arrhizus, Bacillus velezensis, Bacillus subtills, and Bacillus cereus were selected as the fermentation strains for the LSQ. After dual-strain co-fermentation, its pro-digestive enzymes and anti-inflammatory activities were significantly enhanced. Particularly, R. arrhizus/B. subtills fermentation group showed the prominent promotion of the enzymatic activities of amyloglucosidase, cellulase and trypsin, with AC200 values < 1.00 and Max fold increase values of 27.39 ± 0.22, 25.39 ± 0.87 and 48.07 ± 1.84, respectively, and anti-inflammatory activity with an IC₅₀ value of 2.35 ± 0.18 mg/mL. Based on the correlation analysis of differential metabolic profiles and activities, the key pharmacodynamic metabolites were analyzed and validated, such as levomycetin succinatea, β-citrylglutamate, D-glucosaminic acid, nikkomycin and fucose 1-phosphate. Among them, D-glucosaminic acid was positively correlated with the promoting activity of amyloglucosidase, cellulose, enzyme trypsin, pepsin, and the inhibitory activity of NO production, and fucose 1-phosphate and nikkomycin had the prominently positive correlation with the promoting activity of pepsin (p < 0.01). In addition, the docking scores between them and digestive enzyme proteins were all < - 5. A new strategy involving the dual-strain fermentation of LSQ was investigated, and clarified the LSQ fermentation strain-constituent-pharmacological activity correlations, which provides a valuable reference for delving into the LSQ fermentation mechanism.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"52"},"PeriodicalIF":2.0,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction of Corynebacterium glutamicum chassis for preferential utilization of xylose by adaptive laboratory evolution.","authors":"Mingxin Gao, Xi Sun, Yiyang Liu, Tao Chen, Zhiwen Wang","doi":"10.1007/s10529-025-03588-2","DOIUrl":"10.1007/s10529-025-03588-2","url":null,"abstract":"<p><strong>Objectives: </strong>To fully utilize lignocellulosic hydrolysate abundant in xylose content, it is necessary to engineer a Corynebacterium glutamicum strain that can preferentially and efficiently utilize xylose in the presence of glucose/xylose mixtures.</p><p><strong>Results: </strong>C. glutamicum strain CGS15X5-E2 was obtained through metabolic engineering and adaptive laboratory evolution (ALE), which preferentially utilizes xylose and completely consumes it before switching to glucose utilization. A genetically defined chassis strain, CGS15X57, was constructed to switch to glucose consumption only after xylose was depleted based on genome analysis and mutation reconstruction, in which xylose utilization capability was also enhanced. The average xylose consumption rate of CGS15X57 reached 0.833 ± 0.048 g/l/h, which was 28.0% higher than that of the control. Three new beneficial mutations (Cgl1992^{267 insert}, Cgl2948^{G208 T} and Cgl2948^556△C) endow C. glutamicum with rapid growth and efficient xylose utilization phenotypes.</p><p><strong>Conclusions: </strong>A chassis strain of C. glutamicum that preferentially and efficiently utilizes xylose has been obtained, facilitating the full utilization of lignocellulosic hydrolysates and the construction of co-culture systems under glucose/xylose mixed sugar conditions.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"51"},"PeriodicalIF":2.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143975987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advances in microbial and bioelectrochemical strategies for degradation of per- and polyfluoroalkyl substances: mechanisms, limitations, and research opportunities.","authors":"Haoran Yang, Jia Liu","doi":"10.1007/s10529-025-03593-5","DOIUrl":"10.1007/s10529-025-03593-5","url":null,"abstract":"<p><p>Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants characterized by strong carbon-fluorine bonds, making them resistant to conventional degradation methods. Their widespread detection in soil, water, and living organisms, coupled with significant potential health risks, has necessitated the development of effective remediation strategies. This review provides a detailed overview of recent advances in biotechnological approaches for PFAS degradation, with a focus on microbial and bioelectrochemical systems (BESs). Microbial species such as Pseudomonas and Acidimicrobium strains have demonstrated the ability to degrade PFAS under both aerobic and anaerobic conditions. Key enzymes, including dehalogenases and oxygenases, play a critical role in catalyzing the breakdown of PFAS. BESs technologies, including microbial fuel cells (MFCs) and microbial electrolysis cells (MECs), offer innovative solutions by combining microbial activity with electrochemical processes to enhance PFAS removal efficiency. Advanced BESs configurations, such as constructed wetland-MFCs, have further demonstrated the potential for enhanced PFAS removal through electrode adsorption and plant uptake. Despite significant progress, challenges remain, including PFAS toxicity, the complexity of environmental matrices, incomplete mineralization, scalability, and public safety concerns. Addressing these issues will require advancements in genetic engineering to develop robust microbial strains, optimization of BESs configurations, and integration with other advanced treatment technologies like advanced oxidation processes. Additionally, refining environmental factors such as pH, temperature, and the presence of humic substances is crucial for maximizing degradation efficiency. Future research should focus on scaling laboratory successes to field-scale applications, developing real-time monitoring tools for degradation processes, and addressing regulatory concerns. Through continuous innovation, biotechnological solutions offer a promising pathway to sustainable and effective PFAS remediation, addressing both environmental and public health concerns.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"48"},"PeriodicalIF":2.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143961432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cost-effective production of squalene using Yarrowia lipolytica via metabolic engineering and fermentation engineering.","authors":"Wen-Bo Lin, Hong Chen, Ze-Qi Song, Yu-Qing Pan, Peng-Cheng Hu, Xiao-Na Yang, Xiang-Yang Lu, Yun Tian, Hu-Hu Liu","doi":"10.1007/s10529-025-03591-7","DOIUrl":"10.1007/s10529-025-03591-7","url":null,"abstract":"<p><p>Squalene is a triterpene with various biological applications. However, the conventioneer squalene industry is limited by complex extraction processes and environmental pollution, necessitating an environmentally sustainable solution to the increasing demand for squalene. Microbial synthesis is a potentially green and efficient method of producing squalene. Acetyl-CoA is a key precursor of squalene. First, we investigated the effects of enhanced acetyl-CoA supply on squalene production, lipid content, and total fatty acid content in Yarrowia lipolytica. Then, strain YLACLH2 with a squalene production of 232.29 mg/L was obtained by co-overexpressing YlACL2 and YlHMG1. Subsequently, the squalene production of YLACLH2 was increased to 514.33 mg/L by fermentation engineering, optimizing fermentation conditions including temperature, media volume, C/N ratio, shaker flask type and medium type. Finally, we investigated the synthesis efficiency of squalene in Y. lipolytica by acid-hydrolyzed sugarcane molasses (AHM) and waste cooking oil (WCO) as carbon sources with optimized fermentation conditions. This study showed that Y. lipolytica has the potential to produce squalene industrially using low-cost substrates. Our study findings provide reference for engineering Y. lipolytica to produce squalene using low-cost substrates and in an environmentally sustainable manner.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"49"},"PeriodicalIF":2.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143960702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic response of Bacillus spp. to heavy metal stress: pathway alterations and metabolite profiles.","authors":"Xiaowen Yang, Xiaotong Lin, Zhenglin Zhou, Bokun Lin, Xin Liu","doi":"10.1007/s10529-025-03589-1","DOIUrl":"10.1007/s10529-025-03589-1","url":null,"abstract":"<p><p>Heavy metal pollution is a global issue that poses significant risks to ecosystems and human health. Microorganisms offer a promising bioremediation approach due to their ability to mitigate metal-induced metabolic damage in an eco-friendly, efficient, and cost-effective manner. Among them, Gram-positive Bacillus species exhibit a high heavy metal adsorption capacity and secrete metabolites with diverse functional properties. Under heavy metal stress, these metabolites play a crucial role in alleviating metal-induced damage. However, the application of Bacillus metabolites in heavy metal remediation faces challenges, including prolonged treatment durations, the necessity for stable environmental conditions, and specific nutrient requirements.This review summarizes recent research on the effects of heavy metal exposure on the metabolic pathways and metabolites of Bacillus spp., elucidates their role in influencing metal bioavailability and chemical transformations, and explores innovative strategies to enhance the stability of Bacillus-mediated heavy metal remediation. The review aims to provide valuable insights for optimizing bioremediation strategies, facilitating the selection of efficient degrading strains, and advancing the sustainable management of heavy metal contamination.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"50"},"PeriodicalIF":2.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143965644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of lactose based autoinduction for heterologous production of an active [NiFe] hydrogenase in E. coli.","authors":"Francisco de la Fuente-Kratzborn, Qin Fan, Peter Neubauer, Matthias Gimpel","doi":"10.1007/s10529-025-03594-4","DOIUrl":"10.1007/s10529-025-03594-4","url":null,"abstract":"<p><strong>Objectives: </strong>This study aims to assess whether a fed-batch-based auto-induction method can enhance active hydrogenase production, encompassing cofactor formation and space-time yield.</p><p><strong>Results: </strong>The recombinant Escherichia coli strain BQF8RH8, possessing plasmids for assembly and proper maturation of Cupriavidus necator regulatory hydrogenase (RH), was cultivated in the fed-batch like EnPresso B medium with an autoinduction lactose-based system. In contrast to classical IPTG induction previously performed, we obtained active RH demonstrating the feasibility of the process for active hydrogenase production.</p><p><strong>Conclusion: </strong>Our results affirm the viability of the previously developed auto-induction strategy also for a functional hydrogenase with the complex maturation process. This significantly accelerates the process and enhances hydrogenase productivity.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"47"},"PeriodicalIF":2.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12053349/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of CRISPR-Cas9 in microbial cell factories.","authors":"Jinhui Yang, Junyan Song, Zeyu Feng, Yunqi Ma","doi":"10.1007/s10529-025-03592-6","DOIUrl":"10.1007/s10529-025-03592-6","url":null,"abstract":"<p><p>Metabolically engineered bacterial strains are rapidly emerging as a pivotal focus in the biosynthesis of an array of bio-based ingredients. Presently, CRISPR (clustered regularly interspaced short palindromic repeats) and its associated RNA-guided endonuclease (Cas9) are regarded as the most promising tool, having ushered in a transformative advancement in genome editing. Because of CRISPR-Cas9's accuracy and adaptability, metabolic engineers are now able to create novel regulatory systems, optimize pathways more effectively, and make extensive genome-scale alterations. Nevertheless, there are still obstacles to overcome in the application of CRISPR-Cas9 in novel microorganisms, particularly those industrial microorganism hosts that are resistant to traditional genetic manipulation techniques. How to further extend CRISPR-Cas9 to these microorganisms is an urgent problem to be solved. This article first introduces the mechanism and application of CRISPR-Cas9, and then discusses how to optimize CRISPR-Cas9 as a genome editing tool, including how to reduce off-target effects and how to improve targeting efficiency by optimizing design. Through offering a comprehensive perspective on the revolutionary effects of CRISPR-Cas9 in microbial cell factories, we hope to stimulate additional research and development in this exciting area.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"46"},"PeriodicalIF":2.0,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144062072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Creation of a highly stable direct electron transfer-type enzyme sensor by combining a hyperthermophilic dehydrogenase and natural electron mediator.","authors":"Miku Maeno, Haruhiko Sakuraba, Toshihisa Ohshima, Shin-Ichiro Suye, Takenori Satomura","doi":"10.1007/s10529-025-03587-3","DOIUrl":"10.1007/s10529-025-03587-3","url":null,"abstract":"<p><p>This study aimed to address the stability limitations of third-generation biosensors using enzymes from mesophilic organisms, by engineering a stable direct electron transfer (DET)-type dehydrogenase capable of transferring electrons extracted from the substrate to the electrode. A fusion protein combining the mediated electron transfer (MET)-type aldose sugar dehydrogenase from the hyperthermophile Pyrobaculum aerophilum (PaeASD), which cannot transfer electrons generated by enzymatic reactions to the electrode without a mediator, and the natural electron transfer protein cytochrome b₅₆₂ (cyt b₅₆₂) was developed to investigate its potential for the DET reaction. A recombinant protein expression system was established in Escherichia coli to produce the PaeASD-cyt b₅₆₂ fusion protein, which was purified from the soluble fraction of the host cells. Intramolecular electron transfer from pyrroloquinoline quinone (PQQ) to the heme group within the PaeASD-cyt b₅₆₂ fusion protein was investigated using UV-Vis absorption spectroscopy. Upon the addition of glucose, an increase in absorption corresponding to reduced heme molecules was observed, indicating electron transfer from glucose to the heme group in the cyt b₅₆₂ component via PQQ in the PaeASD component. The DET capability of the fusion protein was evaluated using cyclic voltammetry with screen-printed carbon electrodes. A glucose concentration-dependent increase in current response confirmed DET activity. Notably, the fusion protein retained over 80% of its initial current response even after 2 months of storage at 4 °C. The novel robust PaeASD-cyt b₅₆₂ fusion protein demonstrated efficient DET capability, highlighting its high potential for application in the development of third-generation biosensors.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"45"},"PeriodicalIF":2.0,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12011955/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144062079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design and application of expression constructs for FMDV serotype O structural proteins.","authors":"Mostafa R Zaher, Dalia M El-Husseini, Mohamed H El-Husseiny, Azza M El Amir, Naglaa M Hagag, Reham H Tammam","doi":"10.1007/s10529-025-03583-7","DOIUrl":"10.1007/s10529-025-03583-7","url":null,"abstract":"<p><p>Design and validate flexible constructs for recombinant expression of FMDV serotype O structural proteins of the circulating topotypes using newly designed degenerate primers. The designed degenerate primers targeting diverse topotypes enabled the successful amplification of VP0, VP1, and VP3 genes. Integration of the essential transcriptional and translational regulatory elements including T7 promoter, leader g10 sequence, and T7 terminator, as well as ribosome binding site (RBS), start and stop codons, respectively via overlap extension PCR empowered efficient expression of these proteins in E. coli. Cloned constructs expressed the target proteins of expected molecular weights: VP0 (34 kDa), VP1 (24 kDa), and VP3 (22 kDa). SDS-PAGE and Western blotting confirmed high protein yield and purity. This platform demonstrated adaptability for diagnostic and vaccine development applications. The workflow offers a robust tool for producing FMDV structural proteins concerning the circulating strains attempting to improve control measures including diagnosis and vaccinations.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"44"},"PeriodicalIF":2.0,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143964137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction and immunogenicity analysis of a recombinant baculovirus targeting the N protein of SARS-CoV-2.","authors":"Bohan Yu, Xiaoli Mo, Li Sui, Yujia Fang, Xudong Tang, Ping Qian","doi":"10.1007/s10529-025-03576-6","DOIUrl":"10.1007/s10529-025-03576-6","url":null,"abstract":"<p><strong>Objectives: </strong>This study aimed to construct recombinant baculoviruses capable of expressing the nucleocapsid N protein of SARS-CoV-2 and to assess the effects of co-administration with sodium butyrate on its expression and immunogenicity.</p><p><strong>Results: </strong>The recombinant BmNPV expressed green fluorescent protein in BmN cells and N protein in mammalian cells. The addition of sodium butyrate significantly enhanced the expression of N protein in HEK293T cells. Following intramuscular injection of recombinant BmNPV into mice, specific antibodies against the N protein were detectedon day 7, 21, and 35. Co-administration with sodium butyrate (1000 mg/kg body weight) significantly enhanced the immunogenicity of the recombinant virus.</p><p><strong>Conclusions: </strong>Recombinant BmNPV expressing the SARS-CoV2 N protein successfully inducedthe anti-N immunogenicity in mice, providing a solid foundationfor the development of novel subunit vaccines against SARS-CoV-2.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"42"},"PeriodicalIF":2.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143961428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}