Biotechnology Letters最新文献

{"title":"Carrier immobilization and auto-exposition favoring reuse of silyletherase SilE-R from Brassica sp. with high activity and enantiospecificity.","authors":"Lisa Pick, Anna L Schumacher, Elif Öztel, Thorsten Mascher, Marion B Ansorge-Schumacher","doi":"10.1007/s10529-025-03600-9","DOIUrl":"https://doi.org/10.1007/s10529-025-03600-9","url":null,"abstract":"<p><strong>Objectives: </strong>Investigation of immobilization methods promoting the use of silyletherases from Brassica sp. for the efficient and enantiospecific hydrolysis of silyl-protected hydroxyl functions.</p><p><strong>Results: </strong>Different supports for adsorptive and covalent binding of the silyletherase SilE-R as well as exposure of the enzyme on the surface of Bacillus subtilis endospores, so-called SporoBeads, were evaluated. While the highest protein loading of 26 mg enzyme per gram was obtained by adsorptive binding, the best combination of specific activity and enantiospecificity was obtained when SilE-R was exposed on SporoBeads. Protein loading was estimated at 2.6 mg per gram of spore, which was in the same range as after covalent binding to a carrier. In six repeated reaction cycles, SporoBeads exposing SilE-R lost less than 10% of their catalytic activity. The enantiomeric excess could not be increased even with short reaction times, but remained constant over all repeated cycles.</p><p><strong>Conclusion: </strong>The exposure of silyletherases on SporoBeads has been identified as a promising approach for the synthetic application of this novel type of enzyme, although some properties relevant for catalytic applications need to be further improved.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"59"},"PeriodicalIF":2.0,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172454","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":"Heterologous expression of flavohemoglobin in Rhodotorula toruloides led to improved lipid production.","authors":"Shouping Lai, Liting Lyu, Aabid Manzoor Shah, Qiongqiong Chen, Shuang Wang, Hongying Wang, Zongbao Kent Zhao","doi":"10.1007/s10529-025-03597-1","DOIUrl":"https://doi.org/10.1007/s10529-025-03597-1","url":null,"abstract":"<p><p>The red yeast Rhodotorula toruloides emerged as a prominent host for microbial lipid production. It has been known that oxygen supply is a limiting factor for de novo lipid biosynthesis especially under high cell-density culture conditions. Bacterial hemoglobin is an oxygen-binding protein that can actively transport oxygen from the environment to metabolic processes. The aim of the present study was to address the problem of oxygen limitation during R. toruloides culture thereby enhancing cell growth and lipid synthesis. Thus, genes encoding flavohemoglobin (SHb) from Sinorhizobium meliloti and hemoglobin (VHb) from Vitreoscilla sp. were separately integrated into the genome of R. toruloides CGMCC 2.1389 by Agrobacterium-mediated transformation method. It was found that the engineered strain 4S-8 with SHB gene integration produced 49% more lipids under shake-flask culture condition than the parent strain. Under the two-stage culture process, the engineered strain 4S-8 produced 23% and 10% more cell mass than the parent strain and the strain 4V-10 with VHB gene integration, respectively, while the lipid titer of strain 4S-8 was 42% higher than that of the parent strain. These results suggested that heterologous expression of SHb in oleaginous hosts may be applicable for improved production of lipids and cell mass, which should facilitate more efficient conversion of sugars into lipids and fatty-acid derived biofuels.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"58"},"PeriodicalIF":2.0,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141201","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":"Enhancing terpenoid production in Saccharomyces cerevisiae via cell morphology engineering.","authors":"Ke Bi, Siqi Zuo, Jiazhang Lian, Zhinan Xu","doi":"10.1007/s10529-025-03584-6","DOIUrl":"https://doi.org/10.1007/s10529-025-03584-6","url":null,"abstract":"<p><p>There is a growing interest in establishing yeast as a cell factory for terpenoid production. Given the hydrophobicity of many terpenoids, expanding the intracellular storage pool is an effective strategy to improve titers. In this study, the storage capacity was enhanced by increasing yeast cell size through the knockout of EST1 and SSN8. The influence on lycopene and 7-dehydrocholesterol production were then evaluated. The results demonstrated that EST1 and SSN8 deletion resulted in 4.27- and 3.45-fold improvement on lycopene production, and 1.99- and 1.2-fold improvement on 7-dehydrocholesterol production, respectively. These findings suggest cell enlargement as a promising strategy for enhancing terpenoid production in yeast.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"55"},"PeriodicalIF":2.0,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144085742","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":"Improvement of transglutaminase production by controlling the morphology of Streptomyces mobaraensis HVCP-Sm1 with microparticle-enhanced cultivation.","authors":"Ting-Ting Chang, Lu-Yao Bian, Chong Zhang","doi":"10.1007/s10529-025-03598-0","DOIUrl":"https://doi.org/10.1007/s10529-025-03598-0","url":null,"abstract":"<p><strong>Objective: </strong>Transglutaminase (TGase) is widely utilized in food, textile, and medical applications due to its superior capacity to facilitate protein cross-linking. The mechanisms driving high TGase production in Streptomyces mobaraensis require further exploration. \"Microparticle-Enhanced Cultivation\" (MPEC), has been shown to significantly influence the morphology of filamentous microorganisms, thereby impacting secondary metabolism.</p><p><strong>Result: </strong>In the study, CaCO₃ and talc were found to increase the TGase activity of HVCP-Sm1 from 15 U/mL to 29.05 U/mL and 31 U/mL respectively. Moreover, CaCO₃ and talc were observed to reduce the diameters of hyphal particles and elongated hyphae while promoting pro-TGase activation. This research expands the understanding of the role of MPEC in the morphology of filamentous microorganisms and its effects on secondary metabolism.</p><p><strong>Conclusion: </strong>The synergistic effects of group sensing, energy metabolism, and the optimization of mycelial morphology enhanced the secondary metabolism of HVCP-Sm1 and increased TGase enzymatic activity.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"57"},"PeriodicalIF":2.0,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144085744","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":"Plant molecular farming: a promising frontier for orphan drug production.","authors":"Balamurugan Shanmugaraj, Kavin Ravi, Kishore Baladevan","doi":"10.1007/s10529-025-03596-2","DOIUrl":"https://doi.org/10.1007/s10529-025-03596-2","url":null,"abstract":"<p><p>Orphan diseases comprise a range of disorders that individually affect a small percentage of people, but collectively impact millions of people worldwide. Patients with this disorder often face significant challenges in diagnosis, treatment, and access to care due to their rare nature and limited understanding and treatment options. In recent years, significant advancements have been made in the global healthcare in addressing the accessibility of essential treatments and medicines, but still challenges persist particularly related to orphan drugs (to treat rare diseases) in the developing world. The accessibility of orphan drugs remains a major challenge, where patients face barriers such as high costs, limited availability, and inadequate healthcare infrastructure. The high cost associated with orphan drugs presents a barrier to affordability for both patients and healthcare systems, causing disparities in access to life-saving treatments. The molecular farming approach utilizing plant-based production systems for recombinant protein production offers a hope for overcoming barriers to orphan drug access in resource-constrained settings. Molecular farming has the potential to produce a wide range of therapeutic proteins and biologics for the treatment of various rare diseases. The FDA approval of plant-derived proteins for the treatment of Gaucher disease (Elelyso) and Fabry disease (Elfabrio) highlights the potential of plant-based expression systems for the development of suitable drugs targeting niche and orphan diseases. This review examines the potential of the plant system in producing orphan drugs and also highlights the opportunities and challenges related to orphan drug manufacturing.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"56"},"PeriodicalIF":2.0,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144085746","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":"Establishment of a high-throughput scale-down clone screening platform for intensified fed-batch culture of CHO cells.","authors":"Haiyan Luo, Shuai Wang, Collin Chong, Lile Wang, Xiaojun Sun, Qian Guo, Sam Zhang, Xiaoyue Chen, Hang Zhou, Weichang Zhou","doi":"10.1007/s10529-025-03573-9","DOIUrl":"https://doi.org/10.1007/s10529-025-03573-9","url":null,"abstract":"<p><strong>Purpose: </strong>To develop a scale-down clone screening platform for the intensified fed-batch (IFB) process to allow efficient identification of high expressing clones fitting the IFB culture strategy in bioreactor.</p><p><strong>Results: </strong>Three monoclonal antibodies (mAbs) were used in the development and validation of the IFB specific clone screening platform for CHO cells. The IFB platform significantly improved titer levels, achieving an average titer of 8 g/L and the highest titer of 9.6 g/L. With similar cell viability, lactate profile and titer levels, both the spin tube model and the AMBR250^@ bioreactor system were effective in screening clones suitable for IFB process. The addition of aurintricarboxylic acid (ATA) and uridine in the process optimization led to a further increase in expression levels in both systems, achieving the highest titer of 12.2 g/L.</p><p><strong>Conclusion: </strong>This IFB-process specific clone screening serves as an alternative platform for industry application that can increase the effectiveness and efficiency of screening high-expressing CHO cell lines for IFB production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"54"},"PeriodicalIF":2.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143960957","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":"Advancing Yarrowia lipolytica sub-organelle engineering with endogenous mitochondrial targeting sequence.","authors":"Krutika Bhoir, K J Hemavathi, Gunjan Prakash","doi":"10.1007/s10529-025-03590-8","DOIUrl":"https://doi.org/10.1007/s10529-025-03590-8","url":null,"abstract":"<p><strong>Objective: </strong>The aim of the study was identification and validation of an endogenous mitochondrial targeting signal (MTS) sequence of Yarrowia lipolytica, for efficient compartmentalization of a target protein to mitochondria.</p><p><strong>Results: </strong>MTS from citrate synthase of Y. lipolytica (YlCISY-MTS) was identified, isolated and fused with green fluorescent protein (GFP) to direct it to the mitochondrial matrix. The efficiency of localization of GFP to mitochondrial matrix with YlCISY-MTS was compared with currently used MTS from Saccharomyces cerevisiae's cytochrome oxidase subunit IV. Confocal microscopy confirmed the targeted and greater GFP localization, underlining the potential of endogenous YlCISY-MTS for mitochondrial engineering in Y. lipolytica. The availability of endogenous MTS will evade the need of codon optimization of S. cerevisiae MTS for mitochondrial engineering in Y. lipolytica. This is the first report of an endogenous MTS of Y. lipolytica.</p><p><strong>Conclusion: </strong>An endogenous MTS of Y. lipolytica has been identified to facilitate the targeted delivery of a protein in the mitochondria enabling future advancements through leveraging the unique subcellular environment for metabolic engineering applications.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 3","pages":"53"},"PeriodicalIF":2.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143965046","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":"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":"https://doi.org/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":"https://doi.org/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":"https://doi.org/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}