Synthetic and Systems Biotechnology最新文献_第8页

Integrating chemistry knowledge in large language models via prompt engineering 通过提示工程在大型语言模型中整合化学知识

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-07-24 DOI: 10.1016/j.synbio.2024.07.004

Hongxuan Liu , Haoyu Yin , Zhiyao Luo , Xiaonan Wang

引用次数: 0

Liposome-based RNAi delivery in honeybee for inhibiting parasite Nosema ceranae 基于脂质体的 RNAi 在蜜蜂中的传递，用于抑制寄生虫野鹅膏蝇

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-07-18 DOI: 10.1016/j.synbio.2024.07.003

Yue Qi , Chen Wang , Haoyu Lang , Yueyi Wang , Xiaofei Wang , Hao Zheng , Yuan Lu

{"title":"Liposome-based RNAi delivery in honeybee for inhibiting parasite Nosema ceranae","authors":"Yue Qi , Chen Wang , Haoyu Lang , Yueyi Wang , Xiaofei Wang , Hao Zheng , Yuan Lu","doi":"10.1016/j.synbio.2024.07.003","DOIUrl":"10.1016/j.synbio.2024.07.003","url":null,"abstract":"<div>Nosema ceranae, a parasite that parasitizes and reproduces in the gut of honeybees, has become a serious threat to the global apiculture industry. RNA interference (RNAi) technology can be used to inhibit N. ceranae growth by targeting silencing the thioredoxin reductase (TrxR) in N. ceranae. However, suitable carriers are one of the reasons limiting the application of RNAi due to the easy degradation of dsRNA in honeybees. As a vesicle composed of a lipid bilayer, liposomes are a good carrier for nucleic acid delivery, but studies in honeybees are lacking. In this study, liposomes were used for double-stranded RNA (dsRNA) dsTrxR delivery triggering RNAi to inhibit the N. ceranae growth in honeybees. Compared to naked dsTrxR, liposome-dsTrxR reduced N. ceranae numbers in the midgut and partially restored midgut morphology without affecting bee survival and gut microbial composition. The results of this study confirmed that liposomes could effectively protect dsRNA from entering the honeybee gut and provide a reference for using RNAi technology to suppress honeybee pests and diseases.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 853-860"},"PeriodicalIF":4.4,"publicationDate":"2024-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24001017/pdfft?md5=4766286501f4e15858133f504ab98853&pid=1-s2.0-S2405805X24001017-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141949676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative omics directed gene discovery and rewiring for normal temperature-adaptive red pigment synthesis by polar psychrotrophic fungus Geomyces sp. WNF-15A 极性精神营养真菌 Geomyces sp. WNF-15A 在正常温度适应性红色素合成过程中的全息比较基因发现和重新布线

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-07-16 DOI: 10.1016/j.synbio.2024.07.002

Haoyu Long , Jiawei Zhou , Yanna Ren , Jian Lu , Nengfei Wang , Haifeng Liu , Xiangshan Zhou , Menghao Cai

{"title":"Comparative omics directed gene discovery and rewiring for normal temperature-adaptive red pigment synthesis by polar psychrotrophic fungus Geomyces sp. WNF-15A","authors":"Haoyu Long , Jiawei Zhou , Yanna Ren , Jian Lu , Nengfei Wang , Haifeng Liu , Xiangshan Zhou , Menghao Cai","doi":"10.1016/j.synbio.2024.07.002","DOIUrl":"10.1016/j.synbio.2024.07.002","url":null,"abstract":"<div>The Antarctic fungus Geomyces sp. WNF-15A can produce high-quality red pigments (AGRP) with good prospects for the use in food and cosmetic area. However, efficient AGRP synthesis relies on low-temperature and thus limits its industrial development. Here genome sequencing and comparative analysis were performed on the wild-type versus to four mutants derived from natural mutagenesis and transposon insertion mutation. Eleven mutated genes were identified from 2309 SNPs and 256 Indels. A CRISPR-Cas9 gene-editing system was established for functional analysis of these genes. Deficiency of scaffold1.t692 and scaffold2.t704 with unknown functions highly improved AGRP synthesis at all tested temperatures. Of note, the two mutants produced comparable levels of AGRP at 20 °C to the wild-type at 14 °C. They also broke the normal-temperature limitation and effectively synthesized AGRP at 25 °C. Comparative metabolomic analysis revealed that deficiency of scaffold1.t692 improved AGRP synthesis by regulation of global metabolic pathways especially downregulation of the competitive pathways. Knockout of key genes responsible for the differential metabolites confirmed the metabolomic results. This study shows new clues for cold-adaptive regulatory mechanism of polar fungi. It also provides references for exploitation and utilization of psychrotrophic fungal resources.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 842-852"},"PeriodicalIF":4.4,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24001005/pdfft?md5=c8452042e6caff66b2f219f2a7e2a9af&pid=1-s2.0-S2405805X24001005-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141732113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synergistic increase in coproporphyrin III biosynthesis by mitochondrial compartmentalization in engineered Saccharomyces cerevisiae 在工程酿酒酵母中通过线粒体分区协同增加共卟啉 III 的生物合成

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-07-14 DOI: 10.1016/j.synbio.2024.07.001

Qidi Guo , Jiaqi Xu , Jiacun Li , Shuyan Tang , Yuhui Cheng , Bei Gao , Liang-Bin Xiong , Jie Xiong , Feng-Qing Wang , Dong-Zhi Wei

{"title":"Synergistic increase in coproporphyrin III biosynthesis by mitochondrial compartmentalization in engineered Saccharomyces cerevisiae","authors":"Qidi Guo , Jiaqi Xu , Jiacun Li , Shuyan Tang , Yuhui Cheng , Bei Gao , Liang-Bin Xiong , Jie Xiong , Feng-Qing Wang , Dong-Zhi Wei","doi":"10.1016/j.synbio.2024.07.001","DOIUrl":"10.1016/j.synbio.2024.07.001","url":null,"abstract":"<div>Coproporphyrin III (CP III), a natural porphyrin derivative, has extensive applications in the biomedical and material industries. S. cerevisiae has previously been engineered to highly accumulate the CP III precursor 5-aminolevulinic acid (ALA) through the C4 pathway. In this study, a combination of cytoplasmic metabolic engineering and mitochondrial compartmentalization was used to enhance CP III production in S. cerevisiae. By integrating pathway genes into the chromosome, the CP III titer gradually increased to 32.5 ± 0.5 mg/L in shake flask cultivation. Nevertheless, increasing the copy number of pathway genes did not consistently enhance CP III synthesis. Hence, the partial synthesis pathway was compartmentalized in mitochondria to evaluate its effectiveness in increasing CP III production. Subsequently, by superimposing the mitochondrial compartmentalization strategy on cytoplasmic metabolic engineered strains, the CP III titer was increased to 64.3 ± 1.9 mg/L. Furthermore, augmenting antioxidant pathway genes to reduce reactive oxygen species (ROS) levels effectively improved the growth of engineered strains, resulting in a further increase in the CP III titer to 82.9 ± 1.4 mg/L. Fed-batch fermentations in a 5 L bioreactor achieved a titer of 402.8 ± 9.3 mg/L for CP III. This study provides a new perspective on engineered yeast for the microbial production of porphyrins.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 834-841"},"PeriodicalIF":4.4,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000991/pdfft?md5=5fa2900d55ebdd1243e8ea2c347b1c0f&pid=1-s2.0-S2405805X24000991-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141623980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cyanamide-inducible expression of homing nuclease I−SceI for selectable marker removal and promoter characterisation in Saccharomyces cerevisiae 氰胺诱导表达归巢核酸酶 I-SceI，用于在酿酒酵母中去除可选择标记和确定启动子特征

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-28 DOI: 10.1016/j.synbio.2024.06.009

Liam McDonnell , Samuel Evans , Zeyu Lu , Mitch Suchoronczak , Jonah Leighton , Eugene Ordeniza , Blake Ritchie , Nik Valado , Niamh Walsh , James Antoney , Chengqiang Wang , Carlos Horacio Luna-Flores , Colin Scott , Robert Speight , Claudia E. Vickers , Bingyin Peng

{"title":"Cyanamide-inducible expression of homing nuclease I−SceI for selectable marker removal and promoter characterisation in Saccharomyces cerevisiae","authors":"Liam McDonnell , Samuel Evans , Zeyu Lu , Mitch Suchoronczak , Jonah Leighton , Eugene Ordeniza , Blake Ritchie , Nik Valado , Niamh Walsh , James Antoney , Chengqiang Wang , Carlos Horacio Luna-Flores , Colin Scott , Robert Speight , Claudia E. Vickers , Bingyin Peng","doi":"10.1016/j.synbio.2024.06.009","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.009","url":null,"abstract":"<div>In synthetic biology, microbial chassis including yeast Saccharomyces cerevisiae are iteratively engineered with increasing complexity and scale. Wet-lab genetic engineering tools are developed and optimised to facilitate strain construction but are often incompatible with each other due to shared regulatory elements, such as the galactose-inducible (GAL) promoter in S. cerevisiae. Here, we prototyped the cyanamide-induced I−SceI expression, which triggered double-strand DNA breaks (DSBs) for selectable marker removal. We further combined cyanamide-induced I−SceI-mediated DSB and maltose-induced MazF-mediated negative selection for plasmid-free in situ promoter substitution, which simplified the molecular cloning procedure for promoter characterisation. We then characterised three tetracycline-inducible promoters showing differential strength, a non-leaky β-estradiol-inducible promoter, cyanamide-inducible DDI2 promoter, bidirectional MAL32/MAL31 promoters, and five pairs of bidirectional GAL1/GAL10 promoters. Overall, alternative regulatory controls for genome engineering tools can be developed to facilitate genomic engineering for synthetic biology and metabolic engineering applications.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 820-827"},"PeriodicalIF":4.4,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X2400098X/pdfft?md5=a13d7248a6edd9e6adb4e09b95a48e69&pid=1-s2.0-S2405805X2400098X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141542947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advances in stress-tolerance elements for microbial cell factories 微生物细胞工厂耐压元件的研究进展

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-28 DOI: 10.1016/j.synbio.2024.06.008

Zheyi Kuang , Xiaofang Yan , Yanfei Yuan , Ruiqi Wang , Haifan Zhu , Youyang Wang , Jianfeng Li , Jianwen Ye , Haitao Yue , Xiaofeng Yang

{"title":"Advances in stress-tolerance elements for microbial cell factories","authors":"Zheyi Kuang , Xiaofang Yan , Yanfei Yuan , Ruiqi Wang , Haifan Zhu , Youyang Wang , Jianfeng Li , Jianwen Ye , Haitao Yue , Xiaofeng Yang","doi":"10.1016/j.synbio.2024.06.008","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.008","url":null,"abstract":"<div>Microorganisms, particularly extremophiles, have evolved multiple adaptation mechanisms to address diverse stress conditions during survival in unique environments. Their responses to environmental coercion decide not only survival in severe conditions but are also an essential factor determining bioproduction performance. The design of robust cell factories should take the balance of their growing and bioproduction into account. Thus, mining and redesigning stress-tolerance elements to optimize the performance of cell factories under various extreme conditions is necessary. Here, we reviewed several stress-tolerance elements, including acid-tolerant elements, saline-alkali-resistant elements, thermotolerant elements, antioxidant elements, and so on, providing potential materials for the construction of cell factories and the development of synthetic biology. Strategies for mining and redesigning stress-tolerance elements were also discussed. Moreover, several applications of stress-tolerance elements were provided, and perspectives and discussions for potential strategies for screening stress-tolerance elements were made.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 793-808"},"PeriodicalIF":4.4,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000978/pdfft?md5=9e21dff9414b79f288247d1c2878f9d9&pid=1-s2.0-S2405805X24000978-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141542945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhanced triacylglycerol metabolism contributes to the efficient biosynthesis of spinosad in Saccharopolyspora spinosa 三酰甘油新陈代谢的增强促进了刺葡萄孢中刺槐皂苷的高效生物合成

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-25 DOI: 10.1016/j.synbio.2024.06.007

Li Cao, Yangchun Liu, Lin Sun, Zirong Zhu, Danlu Yang, Ziyuan Xia, Duo Jin, Zirui Dai, Jie Rang, Liqiu Xia

{"title":"Enhanced triacylglycerol metabolism contributes to the efficient biosynthesis of spinosad in Saccharopolyspora spinosa","authors":"Li Cao, Yangchun Liu, Lin Sun, Zirong Zhu, Danlu Yang, Ziyuan Xia, Duo Jin, Zirui Dai, Jie Rang, Liqiu Xia","doi":"10.1016/j.synbio.2024.06.007","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.007","url":null,"abstract":"<div>Triacylglycerol (TAG) is crucial for antibiotic biosynthesis derived from Streptomyces, as it serves as an important carbon source. In this study, the supplementation of exogenous TAG led to a 3.92-fold augmentation in spinosad production. The impact of exogenous TAG on the metabolic network of Saccharopolyspora spinosa were deeply analyzed through comparative proteomics. To optimize TAG metabolism and enhance spinosad biosynthesis, the lipase-encoding genes lip886 and lip385 were overexpressed or co-expressed. The results shown that the yield of spinosad was increased by 0.8-fold and 0.4-fold when lip886 and lip385 genes were overexpressed, respectively. Synergistic co-expression of these genes resulted in a 2.29-fold increase in the yield of spinosad. Remarkably, the combined overexpression of lip886 and lip385 in the presence of exogenous TAG elevated spinosad yields by 5.5-fold, led to a drastic increase in spinosad production from 0.036 g/L to 0.234 g/L. This study underscores the modification of intracellular concentrations of free fatty acids (FFAs), short-chain acyl-CoAs, ATP, and NADPH as mechanisms by which exogenous TAG modulates spinosad biosynthesis. Overall, the findings validate the enhancement of TAG catabolism as a beneficial strategy for optimizing spinosad production and provide foundational insights for engineering secondary metabolite biosynthesis pathways in another Streptomyces.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 809-819"},"PeriodicalIF":4.4,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000966/pdfft?md5=c7e671fa9dfff8518fbfe866dcecef77&pid=1-s2.0-S2405805X24000966-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141542946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A systematic discussion and comparison of the construction methods of synthetic microbial community 系统讨论和比较合成微生物群落的构建方法

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-20 DOI: 10.1016/j.synbio.2024.06.006

Chenglong Li, Yanfeng Han, Xiao Zou, Xueqian Zhang, Qingsong Ran, Chunbo Dong

{"title":"A systematic discussion and comparison of the construction methods of synthetic microbial community","authors":"Chenglong Li, Yanfeng Han, Xiao Zou, Xueqian Zhang, Qingsong Ran, Chunbo Dong","doi":"10.1016/j.synbio.2024.06.006","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.006","url":null,"abstract":"<div>Synthetic microbial community has widely concerned in the fields of agriculture, food and environment over the past few years. However, there is little consensus on the method to synthetic microbial community from construction to functional verification. Here, we review the concept, characteristics, history and applications of synthetic microbial community, summarizing several methods for synthetic microbial community construction, such as isolation culture, core microbiome mining, automated design, and gene editing. In addition, we also systematically summarized the design concepts, technological thresholds, and applicable scenarios of various construction methods, and highlighted their advantages and limitations. Ultimately, this review provides four efficient, detailed, easy-to-understand and -follow steps for synthetic microbial community construction, with major implications for agricultural practices, food production, and environmental governance.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 775-783"},"PeriodicalIF":4.4,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000954/pdfft?md5=1ce7685783df1d9189ef567863d52d18&pid=1-s2.0-S2405805X24000954-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141438590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Screening and characterization of integration sites based on CRISPR-Cpf1 in Pichia pastoris 在 Pichia pastoris 中筛选和鉴定基于 CRISPR-Cpf1 的整合位点

IF 4.8 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-18 DOI: 10.1016/j.synbio.2024.06.002

Shupeng Ruan , Yuxin Yang , Xinying Zhang , Guanjuan Luo , Ying Lin , Shuli Liang

{"title":"Screening and characterization of integration sites based on CRISPR-Cpf1 in Pichia pastoris","authors":"Shupeng Ruan , Yuxin Yang , Xinying Zhang , Guanjuan Luo , Ying Lin , Shuli Liang","doi":"10.1016/j.synbio.2024.06.002","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.002","url":null,"abstract":"<div>Pichia pastoris, a methylotrophic yeast, can utilize methanol as a carbon source and energy source to synthesize high-value chemicals, and is an ideal host for biomanufacturing. Constructing the P. pastoris cell factory is somewhat impeded due to the absence of genetic tools for manipulating multi-gene biosynthetic pathways. To broaden its application in the field of metabolic engineering, this study identified and screened 15 novel integration sites in P. pastoris using CRISPR-Cpf1 genome editing technology, with EGFP serving the reporter protein. These integration sites have integration efficiencies of 10–100 % and varying expression strengths, which allow for selection based on the expression levels of genes as needed. Additionally, these integrated sites are applied in the heterologous biosynthesis of P. pastoris, such as the astaxanthin biosynthetic pathway and the carbon dioxide fixation pathway of the Calvin-Benson-Bassham (CBB) cycle. During the three-site integration process, the 8 genes of the CBB cycle were integrated into the genome of P. pastoris. This indicates the potential of these integration sites for integrating large fragments and suggests their successful application in metabolic engineering of P. pastoris. This may lead to improved efficiency of genetic engineering in P. pastoris.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 759-765"},"PeriodicalIF":4.8,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000917/pdfft?md5=bf68a535595642f9f0268209b4e5ebfb&pid=1-s2.0-S2405805X24000917-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141429022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Screening of ent-copalyl diphosphate synthase and metabolic engineering to achieve de novo biosynthesis of ent-copalol in Saccharomyces cerevisiae 筛选ent-copalyl二磷酸合成酶和代谢工程，在酿酒酵母中实现ent-copalol的从头生物合成

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-06-18 DOI: 10.1016/j.synbio.2024.06.005

Shan Li , Shuangshuang Luo , Xinran Yin , Xingying Zhao , Xuyang Wang , Song Gao , Sha Xu , Jian Lu , Jingwen Zhou

{"title":"Screening of ent-copalyl diphosphate synthase and metabolic engineering to achieve de novo biosynthesis of ent-copalol in Saccharomyces cerevisiae","authors":"Shan Li , Shuangshuang Luo , Xinran Yin , Xingying Zhao , Xuyang Wang , Song Gao , Sha Xu , Jian Lu , Jingwen Zhou","doi":"10.1016/j.synbio.2024.06.005","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.06.005","url":null,"abstract":"<div>The diterpene ent-copalol is an important precursor to the synthesis of andrographolide and is found only in green chiretta (Andrographis paniculata). De novo biosynthesis of ent-copalol has not been reported, because the catalytic activity of ent-copalyl diphosphate synthase (CPS) is very low in microorganisms. In order to achieve the biosynthesis of ent-copalol, Saccharomyces cerevisiae was selected as the chassis strain, because its endogenous mevalonate pathway and dephosphorylases could provide natural promotion for the synthesis of ent-copalol. The strain capable of synthesizing diterpene geranylgeranyl pyrophosphate was constructed by strengthening the mevalonate pathway genes and weakening the competing pathway. Five full-length ApCPSs were screened by transcriptome sequencing of A. paniculata and ApCPS2 had the best activity and produced ent-CPP exclusively. The peak area of ent-copalol was increased after the ApCPS2 saturation mutation and its configuration was determined by NMR and ESI-MS detection. By appropriately optimizing acetyl-CoA supply and fusion-expressing key enzymes, 35.6 mg/L ent-copalol was generated. In this study, de novo biosynthesis and identification of ent-copalol were achieved and the highest titer ever reported. It provides a platform strain for the further pathway analysis of andrographolide and derivatives and provides a reference for the synthesis of other pharmaceutical intermediates.</div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"9 4","pages":"Pages 784-792"},"PeriodicalIF":4.4,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000942/pdfft?md5=d092e35682132d8845904ea2be622283&pid=1-s2.0-S2405805X24000942-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141438591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0