Engineering Microbiology最新文献_第7页

Investigating the cellular functions of β-Glucosidases for synthesis of lignocellulose-degrading enzymes in Trichoderma reesei β-葡萄糖苷酶在里氏木霉木质纤维素降解酶合成中的细胞功能研究

Engineering Microbiology Pub Date : 2023-07-05 DOI: 10.1016/j.engmic.2023.100105

Ai–Ping Pang , Haiyan Wang , Yongsheng Luo , Funing Zhang , Fu–Gen Wu , Zhihua Zhou , Zuhong Lu , Fengming Lin

{"title":"Investigating the cellular functions of β-Glucosidases for synthesis of lignocellulose-degrading enzymes in Trichoderma reesei","authors":"Ai–Ping Pang , Haiyan Wang , Yongsheng Luo , Funing Zhang , Fu–Gen Wu , Zhihua Zhou , Zuhong Lu , Fengming Lin","doi":"10.1016/j.engmic.2023.100105","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100105","url":null,"abstract":"<div>β-glucosidases play an important role in the synthesis of cellulase in fungi, but their molecular functions and mechanisms remain unknown. We found that the 10 putative β-glucosidases investigated in Trichoderma reesei facilitate cellulase production, with cel3j being the most crucial. Transcriptional analysis revealed that the most affected biological processes in △cel3j strain were cellulase synthesis, ribosome biogenesis, and RNA polymerases. Moreover, CEL3J was unconventionally transported through the endoplasmic reticulum, bypassing the Golgi apparatus, whereas cel3j overexpression altered cellulase secretion from conventional to unconventional, likely owing to the activated unconventional protein secretion pathway (UPS), as indicated by the upregulation of genes related to UPS. The mTORC1-GRASP55 signaling axis may modulate the unconventional secretion of CEL3J and cellulase. The transcriptional levels of genes associated with DNA replication, the cell cycle, and meiosis were noticeably affected by overexpressing cel3j. These data give new clues for exploring the roles of β-glucosidases and the molecular mechanisms of their unconventional secretion in fungi.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100105"},"PeriodicalIF":0.0,"publicationDate":"2023-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The diverse landscape of AB5-type toxins AB5型毒素的多样性

Engineering Microbiology Pub Date : 2023-06-25 DOI: 10.1016/j.engmic.2023.100104

Paris I. Brown, Adaobi Ojiakor, Antonio J. Chemello, Casey C. Fowler

{"title":"The diverse landscape of AB5-type toxins","authors":"Paris I. Brown, Adaobi Ojiakor, Antonio J. Chemello, Casey C. Fowler","doi":"10.1016/j.engmic.2023.100104","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100104","url":null,"abstract":"<div>AB5-type toxins are a group of secreted protein toxins that are central virulence factors for bacterial pathogens such as Shigella dysenteriae, Vibrio cholerae, Bordetella pertussis, and certain lineages of pathogenic Escherichia coli and Salmonella enterica. AB5 toxins are composed of an active (A) subunit that manipulates host cell biology in complex with a pentameric binding/delivery (B) subunit that mediates the toxin's entry into host cells and its subsequent intracellular trafficking. Broadly speaking, all known AB5-type toxins adopt similar structural architectures and employ similar mechanisms of binding, entering and trafficking within host cells. Despite this, there is a remarkable amount of diversity amongst AB5-type toxins; this includes different toxin families with unrelated activities, as well as variation within families that can have profound functional consequences. In this review, we discuss the diversity that exists amongst characterized AB5-type toxins, with an emphasis on the genetic and functional variability within AB5 toxin families, how this may have evolved, and its impact on human disease.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100104"},"PeriodicalIF":0.0,"publicationDate":"2023-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The influence of the copy number of invader on the fate of bacterial host cells in the antiviral defense by CRISPR-Cas10 DNases CRISPR-Cas10 DNA酶抗病毒防御中入侵者拷贝数对细菌宿主细胞命运的影响

Engineering Microbiology Pub Date : 2023-06-24 DOI: 10.1016/j.engmic.2023.100102

Zhenxiao Yu , Jianan Xu , Yan Zhang , Qunxin She

{"title":"The influence of the copy number of invader on the fate of bacterial host cells in the antiviral defense by CRISPR-Cas10 DNases","authors":"Zhenxiao Yu , Jianan Xu , Yan Zhang , Qunxin She","doi":"10.1016/j.engmic.2023.100102","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100102","url":null,"abstract":"<div>Type III CRISPR-Cas10 systems employ multiple immune activities to defend their hosts against invasion from mobile genetic elements (MGEs), including DNase and cyclic oligoadenylates (cOA) synthesis both of which are hosted by the type-specific protein Cas10. Extensive investigations conducted for the activation of Cas accessory proteins by cOAs have revealed their functions in the type III immunity, but the function of the Cas10 DNase in the same process remains elusive. Here, Lactobacillus delbrueckii subsp. Bulgaricus type III-A (Ld) Csm system, a type III CRISPR system that solely relies on its Cas10 DNase for providing immunity, was employed as a model to investigate the DNase function. Interference assay was conducted in Escherichia coli using two plasmids: pCas carrying the LdCsm system and pTarget producing target RNAs. The former functioned as a de facto “CRISPR host element” while the latter, mimicking an invading MGE. We found that, upon induction of immune responses, the fate of each genetic element was determined by their copy numbers: plasmid of a low copy number was selectively eliminated from the E. coli cells regardless whether it represents a de facto CRISPR host or an invader. Together, we reveal, for the first time, that the immune mechanisms of Cas10 DNases are of two folds: the DNase activity is capable of removing low-copy invaders from infected cells, but it also leads to abortive infection when the invader copy number is high.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100102"},"PeriodicalIF":0.0,"publicationDate":"2023-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advances in the dynamic control of metabolic pathways in Saccharomyces cerevisiae 酿酒酵母代谢途径的动态控制研究进展

Engineering Microbiology Pub Date : 2023-06-21 DOI: 10.1016/j.engmic.2023.100103

Chufan Xiao, Yuyang Pan, Mingtao Huang

引用次数: 1

CRISPR-based gene editing technology and its application in microbial engineering 基于CRISPR的基因编辑技术及其在微生物工程中的应用

Engineering Microbiology Pub Date : 2023-06-20 DOI: 10.1016/j.engmic.2023.100101

Junwei Wei, Yingjun Li

引用次数: 1

Combinatorial metabolic engineering of Saccharomyces cerevisiae for improved production of 7-dehydrocholesterol 酿酒酵母的组合代谢工程改进7-脱氢胆固醇的生产

Engineering Microbiology Pub Date : 2023-06-17 DOI: 10.1016/j.engmic.2023.100100

Yuehao Gu , Shuhui Chen , Xue Jiao , Qi Bian , Lidan Ye , Hongwei Yu

{"title":"Combinatorial metabolic engineering of Saccharomyces cerevisiae for improved production of 7-dehydrocholesterol","authors":"Yuehao Gu , Shuhui Chen , Xue Jiao , Qi Bian , Lidan Ye , Hongwei Yu","doi":"10.1016/j.engmic.2023.100100","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100100","url":null,"abstract":"<div>7-Dehydrocholesterol (7-DHC), a key pharmaceutical intermediate in the production of vitamin D3, has a wide range of applications. To explore fermentative synthesis of 7-DHC, a 7-DHC-producing Saccharomyces cerevisiae strain was constructed by blocking the competitive pathway, eliminating rate-limiting steps, altering global regulation, and pathway compartmentalization. After blocking the competitive pathway by disrupting ERG5 and ERG6 and introducing DHCR24 from Gallus gallus, S. cerevisiae produced 139.72 mg/L (17.04 mg/g dry cell weight, hereafter abbreviated as DCW) 7-DHC. Subsequent alteration of global regulation by deleting ROX1 and overexpressing UPC2-1 increased 7-DHC production to 217.68 mg/L (37.56 mg/g DCW). To remove the accumulated squalene, the post-squalene pathway was strengthened by co-overexpression of PGAL1-driven ERG11 and PGAL10-driven ERG1, which improved 7-DHC titer and yield to 281.73 mg/L and 46.78 mg/g DCW, respectively, and reduced squalene content by 90.12%. We surmised that the sterol precursors in the plasma membrane and peroxisomes may not be accessible to the pathway enzymes, thus we re-localized DHCR24p and Erg2p-GGGGS-Erg3p to the plasma membrane and peroxisomes, boosting 7-DHC production to 357.53 mg/L (63.12 mg/g DCW). Iron supplementation further increased 7-DHC production to 370.68 mg/L in shake flasks and 1.56 g/L in fed-batch fermentation. This study demonstrates the power of global regulation and subcellular relocalization of key enzymes to improve 7-DHC synthesis in yeast.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100100"},"PeriodicalIF":0.0,"publicationDate":"2023-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203818","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Genetic tools for metabolic engineering of Pichia pastoris 毕赤酵母代谢工程的遗传工具

Engineering Microbiology Pub Date : 2023-06-14 DOI: 10.1016/j.engmic.2023.100094

Xiaoyan Wu , Peng Cai , Lun Yao , Yongjin J Zhou

引用次数: 3

Identification of multiple regulatory genes involved in TGase production in Streptomyces mobaraensis DSM 40587 mobaraensis链霉菌DSM 40587中参与TGase产生的多个调控基因的鉴定

Engineering Microbiology Pub Date : 2023-06-10 DOI: 10.1016/j.engmic.2023.100098

Xian Liu , Dan Wang , Yuru Zhang , Xiaoxin Zhuang , Linquan Bai

{"title":"Identification of multiple regulatory genes involved in TGase production in Streptomyces mobaraensis DSM 40587","authors":"Xian Liu , Dan Wang , Yuru Zhang , Xiaoxin Zhuang , Linquan Bai","doi":"10.1016/j.engmic.2023.100098","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100098","url":null,"abstract":"<div>Microbial transglutaminase (TGase) is a protein that is secreted in a mature form and finds wide applications in meat products, tissue scaffold crosslinking, and textile engineering. Streptomyces mobaraensis is the only licensed producer of TGase. However, increasing the production of TGase using metabolic engineering and heterologous expression approaches has encountered challenges in meeting industrial demands. Therefore, it is necessary to identify the regulatory networks involved in TGase biosynthesis to establish a stable and highly efficient TGase cell factory. In this study, we employed a DNA-affinity capture assay and mass spectrometry analysis to discover several transcription factors. Among the candidates, eight were selected and found to impact TGase biosynthesis. Notably, SMDS_4150, an AdpA-family regulator, exhibited a significant influence and was hence named AdpASm. Through electrophoretic mobility shift assays, we determined that AdpASm regulates TGase biosynthesis by directly repressing the transcription of tg and indirectly inhibiting the transcription of SMDS_3961. The latter gene encodes a LytR-family positive regulator of TGase biosynthesis. Additionally, AdpASm exhibited negative regulation of its own transcription. To further enhance TGase production, we combined the overexpression of SMDS_3961 with the repression of SMDS_4150, resulting in a remarkable improvement in TGase titer from 28.67 to 52.0 U/mL, representing an 81.37% increase. This study establishes AdpA as a versatile regulator involved in coordinating enzyme biosynthesis in Streptomyces species. Furthermore, we elucidated a cascaded regulatory network governing TGase production.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100098"},"PeriodicalIF":0.0,"publicationDate":"2023-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli 大肠杆菌谷氨酸脱羧酶系统关键基因的高效共表达产生γ-氨基丁酸

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2023.100077

Lili Yao , Changjiang Lyu , Yuting Wang , Sheng Hu , Weirui Zhao , Hongwei Cao , Jun Huang , Lehe Mei

{"title":"High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli","authors":"Lili Yao , Changjiang Lyu , Yuting Wang , Sheng Hu , Weirui Zhao , Hongwei Cao , Jun Huang , Lehe Mei","doi":"10.1016/j.engmic.2023.100077","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100077","url":null,"abstract":"<div>Biosynthesis of the functional factor γ-aminobutyric acid (GABA) in bacteria involves two key proteins an intracellular glutamate decarboxylase (GadB) and a membrane-bound antiporter (GadC). Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity. In this study, gadBΔC11 of Lactiplantibacillus plantarum and gadCΔC41 of Escherichia coli were inserted into the designed double promoter (PT7lac and PBAD) expression system. Then, E. coli Lemo21(DE3) was chosen as the host to minimize the toxic effects of GadCΔC41 overexpression. Furthermore, a green and high-efficiency GABA synthesis system using dormant engineered Lemo21(DE3) cells as biocatalysts was developed. The total GABA yield reached 829.08 g/L with a 98.7% conversion ratio within 13 h, when engineered E. coli Lemo21(DE3) cells were concentrated to an OD600 of 20 and reused for three cycles in a 3 M L-glutamate solution at 37 °C, which represented the highest GABA productivity ever reported. Overall, expanding the active pH ranges of GadB and GadC toward physiological pH and employing a tunable expression host for membrane-bound GadC production is a promising strategy for high-level GABA biosynthesis in E. coli.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 2","pages":"Article 100077"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49890651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Greener approach to the comprehensive utilization of algal biomass and oil using novel Clostridial fusants and bio-based solvents 利用新型梭状芽孢杆菌融合体和生物基溶剂的藻类生物质和油的绿色综合利用方法

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2022.100068

Asma Fiayaz, Yaser Dahman

{"title":"Greener approach to the comprehensive utilization of algal biomass and oil using novel Clostridial fusants and bio-based solvents","authors":"Asma Fiayaz, Yaser Dahman","doi":"10.1016/j.engmic.2022.100068","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100068","url":null,"abstract":"<div>A greener method has been tested to utilize algal biomass as a feedstock to produce bio-oil in addition to acetone, butanol, and ethanol (ABE) products. Various hydrolysis treatments were used prior to fermentation including combination of thermal, chemical, and enzymatic, which resulted in maximum sugar release of 27.78 g/L. Bio-based terpenes was used instead of common toxic chemicals together with Clostridial fustants to produce bio-alcoholic fuels. Protoplast fusion technique were used to produce the novel Clostridia fusants (C. beijernickii + C. thermocellum and C. acetobutylicum + C. thermocellocum). Fused strains were then subjected to UV radiation for strain enhancement. Final fusansts showed clear improvement in thermal stability and resistance to biobutanol toxicity. Fermentation experiments showed maximum biobutanol final production of 7.98 g/L using CbCt versus 7.39 g/L using CaCt. Oil extraction from virgin algae was tested using a green, bio-based approach using terpenes with ultrasonication and green Bligh and Dyer method, separately. In preliminary study on algal biomass, the combinations of ultrasonication followed by the green Bligh and Dyer have resulted in oil yield of 46.27% (d-limonene) and 39.85% (p-cymene). Oil extraction from an algae sample following fermentation using the combined extraction method resulted in significantly higher oil yield of 65.04%.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 2","pages":"Article 100068"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49890672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1