Biotechnology Journal最新文献_第9页

Development of a biotechnology process for the production of a novel hyperglycosylated long-acting recombinant bovine follicle-stimulating hormone 开发生产新型高糖基化长效重组牛卵泡刺激素的生物技术工艺。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-06-20 DOI: 10.1002/biot.202400260

Javier Villarraza, Sebastián Antuña, María Belén Tardivo, María Celeste Rodríguez, Pablo Uriel Díaz, Ulises Sebastián Notaro, Hugo Héctor Ortega, Claudio Prieto, Natalia Ceaglio

{"title":"Development of a biotechnology process for the production of a novel hyperglycosylated long-acting recombinant bovine follicle-stimulating hormone","authors":"Javier Villarraza, Sebastián Antuña, María Belén Tardivo, María Celeste Rodríguez, Pablo Uriel Díaz, Ulises Sebastián Notaro, Hugo Héctor Ortega, Claudio Prieto, Natalia Ceaglio","doi":"10.1002/biot.202400260","DOIUrl":"10.1002/biot.202400260","url":null,"abstract":"Follicle-stimulating hormone (FSH) is an important protein used for bovine ovarian hyperstimulation in multiple ovulation and embryo transfer technology (MOET). Several attempts to produce bovine FSH (bFSH) in recombinant systems have been reported, nonetheless, up to date, the most commonly used products are partially purified preparations derived from porcine or ovine (pFSH or oFSH) pituitaries. Here we describe the development of a biotechnology process to produce a novel, hyperglycosylated, long-acting recombinant bFSH (LA-rbFSH) by fusing copies of a highly O-glycosylated peptide. LA-rbFSH and a nonmodified version (rbFSH) were produced in suspension CHO cell cultures and purified by IMAC with high purity levels (>99%). LA-rbFSH presented a higher glycosylation degree and sialic acid content than rbFSH. It also demonstrated a notable improvement in pharmacokinetic properties after administration to rats, including a higher concentration in plasma and a significant (seven-fold) reduction in apparent clearance (CLapp). In addition, the in vivo specific bioactivity of LA-rbFSH in rats was 2.4-fold higher compared to rbFSH. These results postulate this new molecule as an attractive substitute for commercially available porcine pituitary-derived products.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unlocking the formate utilization of wild-type Yarrowia lipolytica through adaptive laboratory evolution 通过适应性实验室进化揭示野生型脂溶性亚罗桿菌对甲酸盐的利用。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-06-20 DOI: 10.1002/biot.202400290

Qian Chen, Yunhong Chen, Zeming Hou, Yuyue Ma, Jianfeng Huang, Zhidan Zhang, Yefu Chen, Xue Yang, Yanfei Zhang, Guoping Zhao

{"title":"Unlocking the formate utilization of wild-type Yarrowia lipolytica through adaptive laboratory evolution","authors":"Qian Chen, Yunhong Chen, Zeming Hou, Yuyue Ma, Jianfeng Huang, Zhidan Zhang, Yefu Chen, Xue Yang, Yanfei Zhang, Guoping Zhao","doi":"10.1002/biot.202400290","DOIUrl":"10.1002/biot.202400290","url":null,"abstract":"Synthetic biology is contributing to the advancement of the global net-negative carbon economy, with emphasis on formate as a member of the one-carbon substrate garnering substantial attention. In this study, we employed base editing tools to facilitate adaptive evolution, achieving a formate tolerance of Yarrowia lipolytica to 1 M within 2 months. This effort resulted in two mutant strains, designated as M25-70 and M25-14, both exhibiting significantly enhanced formate utilization capabilities. Transcriptomic analysis revealed the upregulation of nine endogenous genes encoding formate dehydrogenases when cultivated utilizing formate as the sole carbon source. Furthermore, we uncovered the pivotal role of the glyoxylate and threonine-based serine pathway in enhancing glycine supply to promote formate assimilation. The full potential of Y. lipolytica to tolerate and utilize formate establishing the foundation for pyruvate carboxylase-based carbon sequestration pathways. Importantly, this study highlights the existence of a natural formate metabolic pathway in Y. lipolytica.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Functional role of geminivirus encoded proteins in the host: Past and present geminivirus 编码蛋白在宿主体内的功能作用：过去与现在。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-06-20 DOI: 10.1002/biot.202300736

Hira Kamal, Muhammad Mubashar Zafar, Abdul Razzaq, Aqsa Parvaiz, Sezai Ercisli, Fei Qiao, Xuefei Jiang

{"title":"Functional role of geminivirus encoded proteins in the host: Past and present","authors":"Hira Kamal, Muhammad Mubashar Zafar, Abdul Razzaq, Aqsa Parvaiz, Sezai Ercisli, Fei Qiao, Xuefei Jiang","doi":"10.1002/biot.202300736","DOIUrl":"10.1002/biot.202300736","url":null,"abstract":"During plant–pathogen interaction, plant exhibits a strong defense system utilizing diverse groups of proteins to suppress the infection and subsequent establishment of the pathogen. However, in response, pathogens trigger an anti-silencing mechanism to overcome the host defense machinery. Among plant viruses, geminiviruses are the second largest virus family with a worldwide distribution and continue to be production constraints to food, feed, and fiber crops. These viruses are spread by a diverse group of insects, predominantly by whiteflies, and are characterized by a single-stranded DNA (ssDNA) genome coding for four to eight proteins that facilitate viral infection. The most effective means to managing these viruses is through an integrated disease management strategy that includes virus-resistant cultivars, vector management, and cultural practices. Dynamic changes in this virus family enable the species to manipulate their genome organization to respond to external changes in the environment. Therefore, the evolutionary nature of geminiviruses leads to new and novel approaches for developing virus-resistant cultivars and it is essential to study molecular ecology and evolution of geminiviruses. This review summarizes the multifunctionality of each geminivirus-encoded protein. These protein-based interactions trigger the abrupt changes in the host methyl cycle and signaling pathways that turn over protein normal production and impair the plant antiviral defense system. Studying these geminivirus interactions localized at cytoplasm-nucleus could reveal a more clear picture of host–pathogen relation. Data collected from this antagonistic relationship among geminivirus, vector, and its host, will provide extensive knowledge on their virulence mode and diversity with climate change.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular design of controllable recombinant adeno-associated virus (AAV) expression systems for enhanced vector production 可控重组腺相关病毒（AAV）表达系统的分子设计，以提高载体产量。

IF 3.2 3区生物学

Biotechnology Journal Pub Date : 2024-06-20 DOI: 10.1002/biot.202300685

Yusuf B. Johari, Thilo H. Pohle, Jared Whitehead, Joseph M. Scarrott, Ping Liu, Ayda Mayer, David C. James

{"title":"Molecular design of controllable recombinant adeno-associated virus (AAV) expression systems for enhanced vector production","authors":"Yusuf B. Johari, Thilo H. Pohle, Jared Whitehead, Joseph M. Scarrott, Ping Liu, Ayda Mayer, David C. James","doi":"10.1002/biot.202300685","DOIUrl":"10.1002/biot.202300685","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) is the leading vector for the delivery of gene therapies. However, low viral genome (VG) titers are common and the proportion of “full” capsids containing the therapeutic gene payload can be highly variable. The coordinated molecular design of plasmids encoding viral components and Helper functions remains a major challenge for rAAV manufacturing. Here we present the design of improved Rep/Cap and Helper plasmids for rAAV2/8 production, (i) a Rep/Cap expression vector harboring independently controllable rep and cap genes and (ii) an improved Helper plasmid harboring E4 gene deletion variants. First, an optimized Rep/Cap vector utilized a truncated p5 promoter, a p5 cis-regulatory element at the 3′ end in combination with a heterologous promoter to drive Cap expression and an additional copy of the rep52/40 gene to overexpress short Rep proteins. We demonstrate that Rep78 is essential for efficient rAAV2/8 production in HEK293 cells, and a higher ratio of short Rep to long Rep proteins enhances genome packaging. Second, we identified regulators and open reading frames within the Helper plasmid that contribute to increased rAAV2/8 production. While L4-33k/22k is integral to optimal production, the use of E4orf6–6/7 subset significantly enhanced VG titer. Together, an optimal combination of engineered Rep/Cap and Helper plasmid variants increased VG titer by 3.1-fold. This study demonstrates that configuring and controlling the expression of the different AAV genetic elements contributes toward high rAAV production and product quality (full/empty capsid ratio).","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":3.2,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.202300685","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Advanced three-dimensional in vitro liver models to study the activity of anticancer drugs 研究抗癌药物活性的先进三维体外肝脏模型。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202400159

Agnieszka Zuchowska, Sonia Frojdenfal, Maciej Trzaskowski, Elzbieta Jastrzebska

{"title":"Advanced three-dimensional in vitro liver models to study the activity of anticancer drugs","authors":"Agnieszka Zuchowska, Sonia Frojdenfal, Maciej Trzaskowski, Elzbieta Jastrzebska","doi":"10.1002/biot.202400159","DOIUrl":"10.1002/biot.202400159","url":null,"abstract":"The liver is one of the most important organs in the human body. It performs many important functions, including being responsible for the metabolism of most drugs, which is often associated with its drug-induced damage. Currently, there are no ideal pharmacological models that would allow the evaluation of the effect of newly tested drugs on the liver in preclinical studies. Moreover, the influence of hepatic metabolism on the effectiveness of the tested drugs is rarely evaluated. Therefore, in this work we present an advanced model of the liver, which reflects most of the morphologically and metabolically important features of the liver in vivo, namely: three-dimensionality, cellular composition, presence of extracellular matrix, distribution of individual cell types in the structure of the liver model, high urea and albumin synthesis efficiency, high cytochrome p450 activity. In addition, the work, based on the example of commonly used anticancer drugs, shows how important it is to take into account hepatic metabolism in the effective assessment of their impact on the target organ, in this case cancer. In our research, we have shown that the most similar to liver in vivo are 3D cellular aggregates composed of three important liver cells, namely hepatocytes (HepG2), hepatic stellate cells (HSCs), and hepatic sinusoidal endothelial cells (HSECs). Moreover, we showed that the cells in 3D aggregate structure need time (cell–cell interactions) to improve proper liver characteristic. The triculture model additionally showed the greatest ability to metabolize selected anticancer drugs.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

ER stress-induced transcriptional response reveals tolerance genes in yeast ER应激诱导的转录反应揭示了酵母中的耐受基因。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202400082

Jingrong Xie, Chufan Xiao, Yuyang Pan, Songlyu Xue, Mingtao Huang

{"title":"ER stress-induced transcriptional response reveals tolerance genes in yeast","authors":"Jingrong Xie, Chufan Xiao, Yuyang Pan, Songlyu Xue, Mingtao Huang","doi":"10.1002/biot.202400082","DOIUrl":"10.1002/biot.202400082","url":null,"abstract":"Saccharomyces cerevisiae is important for protein secretion studies, yet the complexities of protein synthesis and secretion under endoplasmic reticulum (ER) stress conditions remain not fully understood. ER stress, triggered by alterations in the ER protein folding environment, poses substantial challenges to cells, especially during heterologous protein production. In this study, we used RNA-seq to analyze the transcriptional responses of yeast strains to ER stress induced by reagents such as tunicamycin (Tm) or dithiothreitol (DTT). Our gene expression analysis revealed several crucial genes, such as HMO1 and BIO5, that are involved in ER-stress tolerance. Through metabolic engineering, the best engineered strain R23 with HMO1 overexpression and BIO5 deletion, showed enhanced ER stress tolerance and improved protein folding efficiency, leading to a 2.14-fold increase in α-amylase production under Tm treatment and a 2.04-fold increase in cell density under DTT treatment. Our findings contribute to the understanding of cellular responses to ER stress and provide a basis for further investigations into the mechanisms of ER stress at the cellular level.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of hydrolyzed cheese whey medium for enhanced bacterial cellulose production by Komagataeibacter rhaeticus MSCL 1463 评估水解干酪乳清培养基对提高 Komagataeibacter rhaeticus MSCL 1463 细菌纤维素产量的作用。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202300529

Sergejs Kolesovs, Kristaps Neiberts, Pavels Semjonovs, Sergejs Beluns, Oskars Platnieks, Sergejs Gaidukovs

{"title":"Evaluation of hydrolyzed cheese whey medium for enhanced bacterial cellulose production by Komagataeibacter rhaeticus MSCL 1463","authors":"Sergejs Kolesovs, Kristaps Neiberts, Pavels Semjonovs, Sergejs Beluns, Oskars Platnieks, Sergejs Gaidukovs","doi":"10.1002/biot.202300529","DOIUrl":"10.1002/biot.202300529","url":null,"abstract":"Industrial production of bacterial cellulose (BC) remains challenging due to significant production costs, including the choice of appropriate growth media. This research focuses on optimization of cheese whey (CW) based media for enhanced production of BC. Two modifications were made for CW medium for BC production with Komagataeibacter rhaeticus MSCL 1463. BC production in a medium of enzymatically hydrolyzed CW (final concentration of monosaccharides: glucose 0.13 g L−1, galactose 1.24 g L−1) was significantly enhanced, achieving a yield of 4.95 ± 0.25 g L−1, which markedly surpasses the yields obtained with the standard Hestrin-Schramm (HS) medium containing 20 g L−1 glucose and acid-hydrolyzed CW (final concentration of monosaccharides: glucose 1.15 g L−1, galactose 2.01 g L−1), which yielded 3.29 ± 0.12 g L−1 and 1.01 ± 0.14 g L−1, respectively. We explored the synergistic effects of combining CW with various agricultural by-products (corn steep liquor (CSL), apple juice, and sugar beet molasses). Notably, the supplementation with 15% corn steep liquor significantly enhanced BC productivity, achieving 6.97 ± 0.17 g L−1. A comprehensive analysis of the BC's physical and mechanical properties indicated significant alterations in fiber diameter (62–167 nm), crystallinity index (71.1–85.9%), and specific strength (35–82 MPa × cm3 g−1), as well as changes in the density (1.1–1.4 g cm−3). Hydrolyzed CW medium supplemented by CSL could be used for effective production of BC.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

All-in-one platform: Versatile, Easy, and User-friendly System (VEUS) based on automated and expert-independent antibody immobilization and immunoassay by utilizing customized movement of magnetic particles 一体化平台：多功能、简单易用的系统（VEUS），利用磁性微粒的定制运动，自动固定抗体并进行免疫测定，无需专家操作。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202400074

Jinsik Yoon, Jiyeong Kim, Sujeong Lim, Heelak Choi, Junghyun Bae, Kibeom Kim, Suk-Heung Song, Yoo-Bok Cho, Wook Park, Yong-Gyun Jung

{"title":"All-in-one platform: Versatile, Easy, and User-friendly System (VEUS) based on automated and expert-independent antibody immobilization and immunoassay by utilizing customized movement of magnetic particles","authors":"Jinsik Yoon, Jiyeong Kim, Sujeong Lim, Heelak Choi, Junghyun Bae, Kibeom Kim, Suk-Heung Song, Yoo-Bok Cho, Wook Park, Yong-Gyun Jung","doi":"10.1002/biot.202400074","DOIUrl":"10.1002/biot.202400074","url":null,"abstract":"The ELISA is the most worldwide method for immunoassay. However, the ELISA is losing ground due to low reproducibility of manual experimental processes in both R&D and IVD areas. An automated platform is a good solution, but there are still limitations owning to extremely high cost and requiring large space to set up especially for a small size laboratory. Here, we present a novel all-in-one platform called “VEUS” settable on the laboratory table that offers comprehensive automation of the entire multiplex immunoassay process by exploiting antibody conjugated magnetic particles, quality control and then immunoanalytical reaction, thereby enhancing detection sensitivity and high reproducibility. As a proof of concept, the system exhibits a sensitive LOD of 0.6 and 3.1 pg mL−1 within 1 h run, comparable precision that of molecular diagnostic systems based on PCR method, enabling rapid multiplex diagnosis of Influenza A, Influenza B, and COVID-19 viruses with similar symptoms. Through automation by the all-in-one system, it can be used by novice users, something innovative for immunoassays, relying heavily on user experience. Furthermore, it can contribute to streamline entire immunoassay processes of diverse biomarkers with high reproducibility and convenience in laboratories.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.202400074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 machine learning-based approach for improving plasmid DNA production in Escherichia coli fed-batch fermentations 一种基于机器学习的方法，用于提高大肠杆菌间歇发酵过程中质粒 DNA 的产量。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202400140

Zhixian Xu, Xiaofeng Zhu, Ali Mohsin, Jianfei Guo, Yingping Zhuang, Ju Chu, Meijin Guo, Guan Wang

{"title":"A machine learning-based approach for improving plasmid DNA production in Escherichia coli fed-batch fermentations","authors":"Zhixian Xu, Xiaofeng Zhu, Ali Mohsin, Jianfei Guo, Yingping Zhuang, Ju Chu, Meijin Guo, Guan Wang","doi":"10.1002/biot.202400140","DOIUrl":"10.1002/biot.202400140","url":null,"abstract":"Artificial Intelligence (AI) technology is spearheading a new industrial revolution, which provides ample opportunities for the transformational development of traditional fermentation processes. During plasmid fermentation, traditional subjective process control leads to highly unstable plasmid yields. In this study, a multi-parameter correlation analysis was first performed to discover a dynamic metabolic balance among the oxygen uptake rate, temperature, and plasmid yield, whilst revealing the heating rate and timing as the most important optimization factor for balanced cell growth and plasmid production. Then, based on the acquired on-line parameters as well as outputs of kinetic models constructed for describing process dynamics of biomass concentration, plasmid yield, and substrate concentration, a machine learning (ML) model with Random Forest (RF) as the best machine learning algorithm was established to predict the optimal heating strategy. Finally, the highest plasmid yield and specific productivity of 1167.74 mg L−1 and 8.87 mg L−1/OD600 were achieved with the optimal heating strategy predicted by the RF model in the 50 L bioreactor, respectively, which was 71% and 21% higher than those obtained in the control cultures where a traditional one-step temperature upshift strategy was applied. In addition, this study transformed empirical fermentation process optimization into a more efficient and rational self-optimization method. The methodology employed in this study is equally applicable to predict the regulation of process dynamics for other products, thereby facilitating the potential for furthering the intelligent automation of fermentation processes.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Engineering of the start condensation domain with improved N-decanoyl catalytic activity for daptomycin biosynthesis 工程设计起始缩合结构域，提高 N-癸酰催化活性，促进达托霉素的生物合成。

IF 4.7 3区生物学

Biotechnology Journal Pub Date : 2024-06-19 DOI: 10.1002/biot.202400202

Wenjie Fan, Lyubin Hu, Yu Yang, Panpan Liu, Yan Feng, Ruo-Xu Gu, Qian Liu

{"title":"Engineering of the start condensation domain with improved N-decanoyl catalytic activity for daptomycin biosynthesis","authors":"Wenjie Fan, Lyubin Hu, Yu Yang, Panpan Liu, Yan Feng, Ruo-Xu Gu, Qian Liu","doi":"10.1002/biot.202400202","DOIUrl":"10.1002/biot.202400202","url":null,"abstract":"Daptomycin, a lipopeptide comprising an N-decanoyl fatty acyl chain and a peptide core, is used clinically as an antimicrobial agent. The start condensation domain (dptC1) is an enzyme that catalyzes the lipoinitiation step of the daptomycin synthesis. In this study, we integrated enzymology, protein engineering, and computer simulation to study the substrate selectivity of the start condensation domain (dptC1) and to screen mutants with improved activity for decanoyl loading. Through molecular docking and computer simulation, the fatty acyl substrate channel and the protein–protein interaction interface of dptC1 are analyzed. Key residues at the protein–protein interface between dptC1 and the acyl carrier were mutated, and a single-point mutant showed more than three-folds improved catalytic efficiency of the target n-decanoyl substrate in comparing with the wild type. Moreover, molecular dynamics simulations suggested that mutants with increased catalytic activity may correlated with a more “open” and contracted substrate binding channel. Our work provides a new perspective for the elucidation of lipopeptide natural products biosynthesis, and also provides new resources to enrich its diversity and optimize the production of important components.","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"19 6","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0