Biotechnology Letters最新文献_第5页

Bacillus alcalophilus RecJ (BaRecJ) can drive the adaptive evolution of S. cerevisiae. 嗜钙芽孢杆菌RecJ （BaRecJ）可以驱动酿酒酵母的适应性进化。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-22 DOI: 10.1007/s10529-024-03552-6

Jixiang Shang, Yanchao Zhang, Zongjun Xu, Shouqing Zhang, Zhongtao Sun, Minggang Zheng

引用次数: 0

Optimization and characterization of collagenase KU665299 and its application in effective in-vitro clot digestion. 胶原酶KU665299的优化、表征及其在体外凝块消化中的应用

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-22 DOI: 10.1007/s10529-025-03615-2

Shikha Chauhan, Kriti Kanwar, Deepika Sharma, Harjodh Singh, Deepak Sharma, Vishal Ahuja, Wamik Azmi

{"title":"Optimization and characterization of collagenase KU665299 and its application in effective in-vitro clot digestion.","authors":"Shikha Chauhan, Kriti Kanwar, Deepika Sharma, Harjodh Singh, Deepak Sharma, Vishal Ahuja, Wamik Azmi","doi":"10.1007/s10529-025-03615-2","DOIUrl":"10.1007/s10529-025-03615-2","url":null,"abstract":"Objective: The study employed response surface methodology (RSM) to optimize physicochemical variables for extracellular collagenase production by gram negative bacterial strain Chryseobacterium contaminans KU665299 under submerged fermentation. It is also revealing the ability of collagenase to degrade collagen, main structural protein in human blood.Result: The study successfully enhanced collagenase activity by 1.2 folds through Response Surface Methodology (RSM) and 5.33 folds through purification of enzyme using ammonium sulfate precipitation and DEAE-Sepharose chromatography (specific activity with 538.0 U/mg). SDS-PAGE analysis identified its molecular weight as 32 kDa. Optimal conditions for the enzyme's activity were pH 7.5 and 40 °C. Kinetic studies of collagenase KU665299 revealed specificity for collagen, with Km and Vmax values of 0.059 mg/l and 588.24 µmol/min/mg, respectively. Zinc and calcium ions enhanced activity, while EDTA and DTT strongly inhibited it. The purified collagenase demonstrated remarkable efficiency in digesting blood clots, fully dissolving 1 ml clots within 40 min at 37 °C, showcasing significant thrombolytic potential.Conclusion: The study successfully optimized and characterized a novel collagenase from C. contaminans KU665299, revealing its high specificity, stability, and efficiency in degrading collagen and its promising ability to rapidly digest blood clots for potential thrombolytic properties.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"79"},"PeriodicalIF":2.1,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144688793","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}

引用次数: 0

A single-vector CRISPR/Cas9 system for genome editing and heterologous enzyme secretion in Saccharomyces cerevisiae: a case study on pectate lyase for coffee mucilage removal. 酿酒酵母菌基因组编辑和异源酶分泌的单载体CRISPR/Cas9系统——以果胶裂解酶去除咖啡粘液为例

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-19 DOI: 10.1007/s10529-025-03621-4

La Ho Truc Lam, Nguyen Huynh Ha Nhi, Vo Thi Hoang Lan, Nguyen Van Hau, Nguyen Hieu Nghia

{"title":"A single-vector CRISPR/Cas9 system for genome editing and heterologous enzyme secretion in Saccharomyces cerevisiae: a case study on pectate lyase for coffee mucilage removal.","authors":"La Ho Truc Lam, Nguyen Huynh Ha Nhi, Vo Thi Hoang Lan, Nguyen Van Hau, Nguyen Hieu Nghia","doi":"10.1007/s10529-025-03621-4","DOIUrl":"10.1007/s10529-025-03621-4","url":null,"abstract":"The CRISPR/Cas9 system facilitates precise genome editing in various organisms. In this study, a single-vector CRISPR/Cas9 system was developed for Saccharomyces cerevisiae, employing a type II Cas9 enzyme from Streptococcus pyogenes and a single-guide RNA cassette targeting CAN1.Y locus on chromosome V. This system is broadly applicable across yeast strains, as it utilizes G418 selection, eliminating the need for auxotrophic markers. The efficiency of the CRISPR/Cas9 system was demonstrated, with editing efficiencies ranging from 70 to 100%. This system was utilized to integrate a cassette encoding secretory pectate lyase (PL) from Bacillus subtilis 168 into the yeast genome. The engineered S. cerevisiae strain secreted active PL, which exhibited pectin-degrading activity characterized by significant reductions in residual pectin and increased production of reducing sugars. Since pectin constitutes a major component of coffee mucilage, the secreted PL was applied to coffee beans for mucilage removal. The treated beans presented noticeably reduced residual mucilage, a purer green color, and decreased viscosity. These findings suggest the potential of the engineered S. cerevisiae strain for applications in coffee processing, particularly in efficient mucilage removal.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"78"},"PeriodicalIF":2.1,"publicationDate":"2025-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144666999","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}

引用次数: 0

A novel method for generating baculovirus bacmids using EGFP-mediated purification and linearization. 利用egfp介导的纯化和线性化生成杆状病毒毒株的新方法。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-17 DOI: 10.1007/s10529-025-03619-y

Wujie Su, Haoyi Gu, Xiaoxia Zhang, Wenbing Wang, Fanchi Li, Bing Li

{"title":"A novel method for generating baculovirus bacmids using EGFP-mediated purification and linearization.","authors":"Wujie Su, Haoyi Gu, Xiaoxia Zhang, Wenbing Wang, Fanchi Li, Bing Li","doi":"10.1007/s10529-025-03619-y","DOIUrl":"10.1007/s10529-025-03619-y","url":null,"abstract":"Baculovirus bacmids have been widely used in over-expression and gene deletion. Traditionally, baculovirus bacmids are developed by inserting an 8.6 kbp bacterial DNA cassette into baculovirus genomes either through homologous recombination in cultured cells or via in vitro cloning. In this study, by introducing Bsu36i-attached egfp to the 8.6 kbp bacterial DNA cassette, we develop a novel method for generating baculovirus bacmids. An 11.6 kbp bacterial DNA cassette containing the introduced egfp was used to generate an intermediate bacmid. With the EGFP reporter, purification was performed in cultured cells, increasing the proportions of recombinants. The intermediate bacmid containing the 11.6 kbp bacterial DNA cassette was obtained by transforming DH10B competent cells with viral DNA after 3 rounds of purification. The intermediate bacmid DNA was linearized by digestion with Bsu36i and then was co-transfected with the PCR-amplified 8.6 kbp bacterial cassette into BmN cells, where homologous recombination occurred between them. The final BmNPV bacmid was obtained by transforming DH10B competent cells with viral DNA. Capable of increasing the proportions of recombinants via purification and linearization, this method has great potential to be used for bacmid generation for baculoviruses, especially those that are not capable of producing high titers of viruses.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"77"},"PeriodicalIF":2.1,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144648424","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}

引用次数: 0

A novel P450 enzyme assay utilizing an NADP⁺-based biosensor. 利用NADP+为基础的生物传感器的新型P450酶测定。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-16 DOI: 10.1007/s10529-025-03599-z

Sifan Shangguan, Taichang Wang, Di Zhao, Guobin Zhang, Yisang Zhang, Ruiming Wang, Junqing Wang, Jing Su

{"title":"A novel P450 enzyme assay utilizing an NADP+-based biosensor.","authors":"Sifan Shangguan, Taichang Wang, Di Zhao, Guobin Zhang, Yisang Zhang, Ruiming Wang, Junqing Wang, Jing Su","doi":"10.1007/s10529-025-03599-z","DOIUrl":"10.1007/s10529-025-03599-z","url":null,"abstract":"Purpose: High-throughput screening methods for cytochrome P450 enzymes (P450s), such as colorimetric, mass spectrometric, and fluorescence-based assays, often face limitations in throughput, real-time monitoring, and versatility.Methods: To address these challenges, we developed a novel biosensor leveraging glucose-6-phosphate dehydrogenase and Bimolecular Fluorescence Complementation for real-time monitoring of intracellular NADP+ levels, enabling P450 activity detection. The sensor was applied to monitor P450 activity by tracking intracellular NADP+ dynamics, as P450s catalyze diverse substrate reactions and convert NADPH to NADP+ via their electron transport system. To enhance detection precision, intracellular NADP+ synthesis was reduced by knocking down NADPH-dependent aldehyde reductase (YqhD), minimizing background fluorescence interference.Results: The sensor exhibited a linear NADP+ detection range of 1 μM to 10 mM, suitable for P450 assays. The sensor's performance was validated by comparing P450 activities in engineered strains with traditional gas chromatography.Conclusion: The developed biosensor demonstrates its potential as a robust, real-time screening tool for P450 enzyme studies.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"76"},"PeriodicalIF":2.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641711","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}

引用次数: 0

Oral administration of Bacillus subtilis spores expressing Staphylococcus aureus IsdB induces mucosal immune responses in mice. 口服表达金黄色葡萄球菌IsdB的枯草芽孢杆菌孢子可诱导小鼠粘膜免疫反应。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-14 DOI: 10.1007/s10529-025-03620-5

Nhi N Y Nguyen, Lan Duong, Tam Th Nguyen, Uyen T T Nguyen, Vy Nguyen Thuy Pham, Hoang Duc Nguyen

{"title":"Oral administration of Bacillus subtilis spores expressing Staphylococcus aureus IsdB induces mucosal immune responses in mice.","authors":"Nhi N Y Nguyen, Lan Duong, Tam Th Nguyen, Uyen T T Nguyen, Vy Nguyen Thuy Pham, Hoang Duc Nguyen","doi":"10.1007/s10529-025-03620-5","DOIUrl":"10.1007/s10529-025-03620-5","url":null,"abstract":"Purpose: Bacillus subtilis spores, known for their durability and well-developed genetic manipulation tools, show promise for oral vaccine delivery. Staphylococcus aureus is a major global health concern due to multidrug resistance and lack of a vaccine. This study explored the potential of B. subtilis spores engineered to express IsdB, a key S. aureus iron acquisition protein, on their surface and evaluated the immunogenic effect of recombinant spores through oral administration in mice.Method: B. subtilis spores were engineered to display a key S. aureus protein, IsdB, fused with spore coat protein, CotB, and confirmed by PCR. Western blotting, sporeELISA, and immunofluorescence microscopy verified the surface expression of IsdB. The engineered BsHT2377 spores were orally administered to Swiss mice at two different doses, and antibody levels in both serum and feces were measured using ELISA.Result: PCR confirmed the targeted integration of the isdB gene into B. subtilis, generating the strain BsHT2377. Western blotting, sporeELISA, and immunofluorescence microscopy validated IsdB surface display on BsHT2377 spores. Oral administration triggered a gut-localised immune response in mice, with significantly elevated fecal IgA but no substantial increase in serum IgG.Conclusion: This study engineered a novel B. subtilis strain, BsHT2377, that displays the S. aureus IsdB protein on its spore surface. Oral administration in mice significantly increased fecal IgA, indicating a mucosal immune response. These findings highlight the potential of B. subtilis spores as oral vaccine carriers and offer insights to support the optimization of future vaccine designs.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"75"},"PeriodicalIF":2.1,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144636051","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}

引用次数: 0

High throughput screening and metabolic engineering of Saccharopolyspora spinosa for spinosad production. 棘糖多孢子菌的高通量筛选及代谢工程研究。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-09 DOI: 10.1007/s10529-025-03617-0

Jiaxin Du, Jiale Zhang, Chen Yang, Chuanbo Zhang, Wenyu Lu

{"title":"High throughput screening and metabolic engineering of Saccharopolyspora spinosa for spinosad production.","authors":"Jiaxin Du, Jiale Zhang, Chen Yang, Chuanbo Zhang, Wenyu Lu","doi":"10.1007/s10529-025-03617-0","DOIUrl":"10.1007/s10529-025-03617-0","url":null,"abstract":"The macrolide antibiotics spinosad, synthesized by Saccharopolyspora spinosa, is a highly effective, environmentally-friendly insecticide. However, due to poor fermentation performance and difficulties in engineering S. spinosa strains, the production cost of spinosad is still high, which restricts its industrial application. The industrial strains used for antibiotic production primarily originate from mutagenic screening, whereas traditional screening methods are time-consuming and laborious. Here, an in vitro detection method for spinosad was established to accelerate the breeding of mutated strains of S. spinosa. Firstly, a broad substrate promiscuity glycosyltransferase OleD from Streptomyces antibioticus was selected and employed for the detection of pseudoaglycone (PSA), which serves as the precursor compound for spinosad, through the utilization of colorimetric reactions coupled with glycosylation. Subsequently, the in vitro PSA detection system was optimized and applied for S. spinosa high throughput screening. The final selected mutant strain DUA15 was obtained and showed a 0.80-fold and 0.66-fold increase in spinosad and PSA production compared to the original strain, respectively. Furthermore, genetic engineering technology was combined to obtain the engineered strain D15-102, which showed a 2.9-fold increase in spinosad production compared to the original strain.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"74"},"PeriodicalIF":2.1,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590374","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}

引用次数: 0

Structural characterization of novel extracellular polymer substances from an Alternaria sp. 17463 and their impact on soil metabolism. Alternaria sp. 17463新型胞外聚合物的结构特征及其对土壤代谢的影响。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-08 DOI: 10.1007/s10529-025-03616-1

Yinli Bi, Hai Tan, Shishuang Zhang, Dongdong Wang, Jing Zhao

{"title":"Structural characterization of novel extracellular polymer substances from an Alternaria sp. 17463 and their impact on soil metabolism.","authors":"Yinli Bi, Hai Tan, Shishuang Zhang, Dongdong Wang, Jing Zhao","doi":"10.1007/s10529-025-03616-1","DOIUrl":"10.1007/s10529-025-03616-1","url":null,"abstract":"Dark septate endophytes (DSE) are widely used in ecological restoration. In this study, we first discovered that the metabolic products of the DSE strain Alternaria sp. 17463 contain extracellular polymeric substances (EPS). This study aimed to optimize the culture conditions, characterize the structural composition of EPS, and evaluate its effects on soil improvement. The ethanol precipitation method was used to extract EPS from the metabolites of Alternaria sp. 17463, increasing the yield 4.48-fold to 6.96 g L⁻1 through optimized cultivation conditions. The molecular mass of EPS was 533.754 kDa, with an exopolysaccharide content of up to 77.8% and a protein content of 8.4%. The EPS were mainly composed of glucan units T)-Glc p-(1 → and → 4)-Glc p-(1 → , with minor fractions of mannose and galactose. By acting as a cementing agent, EPS strengthened the linkages between soil particles. At an optimal concentration, it significantly enhanced the activity of extracellular enzymes involved in carbon, nitrogen, and phosphorus metabolism, thereby improving soil nutrient transformation and cycling. This also promoted changes in the metabolism of small molecules in the soil. This research reveals why this strain can accelerate ecological restoration and lays the foundation for further exploration of its metabolic products and potential applications in this field, particularly in open-pit dump reclamation and agricultural soil improvement.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"73"},"PeriodicalIF":2.1,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590375","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}

引用次数: 0

High-density heterotrophic cultivation of a cell-wall-deficient Chlamydomonas reinhardtii strain by fed-batch strategy. 一株细胞壁缺陷莱茵衣藻高密度异养培养的补料分批策略。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-07-03 DOI: 10.1007/s10529-025-03614-3

Kaidi Zhou, Tao Yu, Minxi Wan, Zhengxu Bao, Wenming Bai, Weiliang Wang, Yuanguang Li

{"title":"High-density heterotrophic cultivation of a cell-wall-deficient Chlamydomonas reinhardtii strain by fed-batch strategy.","authors":"Kaidi Zhou, Tao Yu, Minxi Wan, Zhengxu Bao, Wenming Bai, Weiliang Wang, Yuanguang Li","doi":"10.1007/s10529-025-03614-3","DOIUrl":"10.1007/s10529-025-03614-3","url":null,"abstract":"Chlamydomonas reinhardtii CW15, a cell-wall-deficient strain, is a widely used chassis in microalgal genetic engineering. However, its low cell density under autotrophic and mixotrophic cultivation restricts industrial scalability. This study aimed to establish a cost-effective, high-density heterotrophic cultivation process for C. reinhardtii CW15. Initially, an optimized AP medium (Tris-free TAP with K2HPO4 as the sole phosphorus source) significantly reduced medium costs while achieving a cell density of 0.446 g/L. Subsequently, a fed-batch process in a 5 L fermentor yielded a record cell density of 22.1 g/L (growth rate: 94.6 mg/L·h) at 233 h by optimizing nutrient supplementation. Scaling up to a 50 L fermentor revealed challenges in ammonia volatilization and salinity accumulation during sterilization. These were resolved by substituting the nitrogen source with ammonia water and supplementing ammonium acetate as a dual nitrogen and partial carbon source, ultimately achieving 30.2 g/L (140 mg/L·h) after 215 h-the highest reported density for cell-wall-deficient C. reinhardtii. This work provides an efficient and scalable cultivation strategy, facilitating its industrial and biotechnological applications.","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"47 4","pages":"72"},"PeriodicalIF":2.1,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558930","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}

引用次数: 0

Role of microbial pesticides in pest control for lepidopteran: research status and prospect. 微生物农药在鳞翅目害虫防治中的作用：研究现状与展望。

IF 2.1 4区生物学

Biotechnology Letters Pub Date : 2025-06-30 DOI: 10.1007/s10529-025-03613-4

LiLi Wei, Yiqing He, Juan Du, Yuejun Fu

引用次数: 0