Engineering Microbiology最新文献_第8页

Global regulation of fungal secondary metabolism in Trichoderma reesei by the transcription factor Ypr1, as revealed by transcriptome analysis 转录组分析揭示了转录因子Ypr1对里氏木霉真菌次生代谢的全局调控

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2022.100065

Jie Yang, Jia-Xiang Li, Fei Zhang, Xin-Qing Zhao

{"title":"Global regulation of fungal secondary metabolism in Trichoderma reesei by the transcription factor Ypr1, as revealed by transcriptome analysis","authors":"Jie Yang, Jia-Xiang Li, Fei Zhang, Xin-Qing Zhao","doi":"10.1016/j.engmic.2022.100065","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100065","url":null,"abstract":"<div>Trichoderma reesei Rut-C-30 is a well-known robust producer of cellulolytic enzymes, which are used to degrade lignocellulosic biomass for the sustainable production of biofuels and biochemicals. However, studies of its secondary metabolism and regulation remain scarce. Ypr1 was previously described as a regulator of the biosynthesis of the yellow pigment sorbicillin (a bioactive agent with great pharmaceutical interest) in T. reesei and several other fungi. However, the manner in which this regulator affects global gene transcription has not been explored. In this study, we report the effect of Ypr1 on the regulation of both the secondary and primary metabolism of T. reesei Rut-C30. A global gene transcription profile was obtained using a comparative transcriptomic analysis of the wild-type strain T. reesei Rut-C-30 and its ypr1 deletion mutant. The results of this analysis suggest that, in addition to its role in regulating sorbicillin and the major extracellular (hemi)cellulases, Ypr1 also affects the transcription of genes encoding several other secondary metabolites. Although the primary metabolism of T. reesei ∆ypr1 became less active compared with that of T. reesei Rut-C-30, several gene clusters involved in its secondary metabolism were activated, such as the gene clusters for the biosynthesis of specific polyketides and non-ribosomal peptides, together with the “sorbicillinoid–cellulase” super cluster, indicating that specific secondary metabolites and cellulases may be co-regulated in T. reesei Rut-C-30. The results presented in this study may benefit the development of genetic engineering strategies for the production of sorbicillin by T. reesei Rut-C-30, and provide insights for enhancing sorbicillin production in other filamentous fungal producers.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 2","pages":"Article 100065"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49890673","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

Advances on the biosynthesis of pyridine rings 吡啶环生物合成研究进展

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2022.100064

Carolina E. Portero , Yong Han , M. Raquel Marchán-Rivadeneira

引用次数: 0

A highly selective cell-based fluorescent biosensor for genistein detection 染料木黄酮检测的高选择性细胞荧光生物传感器

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2023.100078

Lucy Fang-I Chao , Dany Liu , Verena Siewers

{"title":"A highly selective cell-based fluorescent biosensor for genistein detection","authors":"Lucy Fang-I Chao , Dany Liu , Verena Siewers","doi":"10.1016/j.engmic.2023.100078","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100078","url":null,"abstract":"<div>Genistein, an isoflavone found mainly in legumes, has been shown to have numerous health benefits for humans. Therefore, there is substantial interest in producing it using microbial cell factories. To aid in screening for high genistein producing microbial strains, a cell-based biosensor for genistein was developed by repurposing the Gal4DBD-ERα-VP16 (GEV) transcriptional activator in Saccharomyces cerevisiae. In the presence of genistein, the GEV sensor protein binds to the GAL1 promoter and activates transcription of a downstream GFP reporter. The performance of the biosensor, as measured by fold difference in GFP signal intensity after external genistein induction, was improved by engineering the sensor protein, its promoter and the reporter promoter. Biosensor performance increased when the weak promoter REV1p was used to drive GEV sensor gene expression and the VP16 transactivating domain on GEV was replaced with the tripartite VPR transactivator that had its NLS removed. The biosensor performance further improved when the binding sites for the inhibitor Mig1 were removed from and two additional Gal4p binding sites were added to the reporter promoter. After genistein induction, our improved biosensor output a GFP signal that was 20 times higher compared to the uninduced state. Out of the 8 flavonoids tested, the improved biosensor responded only to genistein and in a somewhat linear manner. The improved biosensor also responded to genistein produced in vivo, with the GFP reporter intensity directly proportional to intracellular genistein concentration. When combined with fluorescence-based cell sorting technology, this biosensor could facilitate high-throughput screening of a genistein-producing yeast cell factory.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 2","pages":"Article 100078"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49890676","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

Fungal arginine-containing cyclodipeptide synthases are finally revealed 真菌含精氨酸环二肽合成酶的最终揭示

Engineering Microbiology Pub Date : 2023-06-01 DOI: 10.1016/j.engmic.2023.100080

Hang Li

引用次数: 0

Yeast surface display of leech hyaluronidase for the industrial production of hyaluronic acid oligosaccharides 水蛭透明质酸酶的酵母表面展示用于透明质酸低聚糖的工业化生产

Engineering Microbiology Pub Date : 2023-04-05 DOI: 10.1016/j.engmic.2023.100086

Lizhi Liao , Hao Huang , Yang Wang , Guocheng Du , Zhen Kang

{"title":"Yeast surface display of leech hyaluronidase for the industrial production of hyaluronic acid oligosaccharides","authors":"Lizhi Liao , Hao Huang , Yang Wang , Guocheng Du , Zhen Kang","doi":"10.1016/j.engmic.2023.100086","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100086","url":null,"abstract":"<div>Leech hyaluronidase (LHyal) is a hyperactive hyaluronic acid (HA) hydrolase that belongs to the hyaluronoglucuronidase family. Traditionally, LHyal is extracted from the heads of leeches, but the recent development of the Pichia pastoris recombinant LHyal expression method permitted the industrial production of size-specific HA oligosaccharides. However, at present LHyal expressed by recombinant yeast strains requires laborious protein purification steps. Moreover, the enzyme is deactivated and removed after single use. To solve this problem, we developed a recyclable LHyal biocatalyst using a yeast surface display (YSD) system. After screening and characterization, we found that the cell wall protein Sed1p displayed stronger anchoring to the P. pastoris cell wall than other cell wall proteins. By optimizing the type and length of the linkers between LHyal and Sed1p, we increased the activity of enzymes displayed on the P. pastoris cell wall by 50.34% in flask cultures. LHyal-(GGGS)6-Sed1p activity further increased to 3.58 × 105 U mL−1 in fed-batch cultivation in a 5 L bioreactor. Enzymatic property analysis results revealed that the displayed LHyal-(GGGS)6-Sed1p generated the same oligosaccharides but exhibited higher thermal stability than free LHyal enzyme. Moreover, displayed LHyal-(GGGS)6-Sed1p could be recovered easily from HA hydrolysis solutions via low-speed centrifugation and could be reused at least 5 times. YSD of LHyal not only increased the utilization efficiency of the enzyme but also simplified the purification process for HA oligosaccharides. Thus, this study provides an alternative approach for the industrial preparation of LHyal and HA oligosaccharides.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100086"},"PeriodicalIF":0.0,"publicationDate":"2023-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203817","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: 3

A strategy to enhance the insecticidal potency of Vip3Aa by introducing additional cleavage sites to increase its proteolytic activation efficiency 一种通过引入额外的切割位点来提高Vip3Aa蛋白水解激活效率来提高其杀虫效力的策略

Engineering Microbiology Pub Date : 2023-03-17 DOI: 10.1016/j.engmic.2023.100083

Kun Jiang , Zhe Chen , Yiting Shi , Yuanrong Zang , Chengbin Shang , Xi Huang , Jiahe Zang , Zhudong Bai , Xuyao Jiao , Jun Cai , Xiang Gao

{"title":"A strategy to enhance the insecticidal potency of Vip3Aa by introducing additional cleavage sites to increase its proteolytic activation efficiency","authors":"Kun Jiang , Zhe Chen , Yiting Shi , Yuanrong Zang , Chengbin Shang , Xi Huang , Jiahe Zang , Zhudong Bai , Xuyao Jiao , Jun Cai , Xiang Gao","doi":"10.1016/j.engmic.2023.100083","DOIUrl":"https://doi.org/10.1016/j.engmic.2023.100083","url":null,"abstract":"<div>Microbially derived, protein-based biopesticides have become a vital element in pest management strategies. Vip3 family proteins from Bacillus thuringiensis have distinct characteristics from known insecticidal Cry toxins and show efficient insecticidal activity against several detrimental lepidopteran pests. They are considered to be a promising toxic candidate for the management of various detrimental pests. In this study, we found that in addition to the preliminary digestion sites lysine, there are multiple cleavage activation sites in the linker region between domain I (DI) and DII of Vip3Aa. We further demonstrated that by adding more cleavage sites between DI and DII of Vip3Aa, its proteolysis efficiency by midgut proteases can be significantly increased, and correspondingly enhance its insecticidal activity against Spodoptera frugiperda and Helicoverpa armigera larvae. Our study promotes the understanding of the insecticidal mechanism of Vip3 proteins and illustrates an easily implementable strategy to increase the insecticidal potency of Vip3Aa. This facilitates their potential future development and efficient application for sustainable agriculture.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 4","pages":"Article 100083"},"PeriodicalIF":0.0,"publicationDate":"2023-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203841","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

Oxidative modification of free-standing amino acids by Fe(II)/αKG-dependent oxygenases Fe（II）/αKG依赖性加氧酶对独立氨基酸的氧化修饰

Engineering Microbiology Pub Date : 2023-03-01 DOI: 10.1016/j.engmic.2022.100062

Hui Tao , Ikuro Abe

{"title":"Oxidative modification of free-standing amino acids by Fe(II)/αKG-dependent oxygenases","authors":"Hui Tao , Ikuro Abe","doi":"10.1016/j.engmic.2022.100062","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100062","url":null,"abstract":"<div>Fe(II)/α-ketoglutarate (αKG)-dependent oxygenases catalyze the oxidative modification of various molecules, from DNA, RNA, and proteins to primary and secondary metabolites. They also catalyze a variety of biochemical reactions, including hydroxylation, halogenation, desaturation, epoxidation, cyclization, peroxidation, epimerization, and rearrangement. Given the versatile catalytic capability of such oxygenases, numerous studies have been conducted to characterize their functions and elucidate their structure–function relationships over the past few decades. Amino acids, particularly nonproteinogenic amino acids, are considered as important building blocks for chemical synthesis and components for natural product biosynthesis. In addition, the Fe(II)/αKG-dependent oxygenase superfamily includes important enzymes for generating amino acid derivatives, as they efficiently modify various free-standing amino acids. The recent discovery of new Fe(II)/αKG-dependent oxygenases and the repurposing of known enzymes in this superfamily have promoted the generation of useful amino acid derivatives. Therefore, this study will focus on the recent progress achieved from 2019 to 2022 to provide a clear view of the mechanism by which these enzymes have expanded the repertoire of free amino acid oxidative modifications.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 1","pages":"Article 100062"},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203892","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

Two-component system GacS/GacA, a global response regulator of bacterial physiological behaviors 双组分系统GacS/GacA，细菌生理行为的全局反应调节因子

Engineering Microbiology Pub Date : 2023-03-01 DOI: 10.1016/j.engmic.2022.100051

Huihui Song , Yuying Li , Yan Wang

{"title":"Two-component system GacS/GacA, a global response regulator of bacterial physiological behaviors","authors":"Huihui Song , Yuying Li , Yan Wang","doi":"10.1016/j.engmic.2022.100051","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100051","url":null,"abstract":"<div>The signal transduction system of microorganisms helps them adapt to changes in their complex living environment. Two-component system (TCS) is a representative signal transduction system that plays a crucial role in regulating cellular communication and secondary metabolism. In Gram-negative bacteria, an unorthodox TCS consisting of histidine kinase protein GacS (initially called LemA) and response regulatory protein GacA is widespread. It mainly regulates various physiological activities and behaviors of bacteria, such as quorum sensing, secondary metabolism, biofilm formation and motility, through the Gac/Rsm (Regulator of secondary metabolism) signaling cascade pathway. The global regulatory ability of GacS/GacA in cell physiological activities makes it a potential research entry point for developing natural products and addressing antibiotic resistance. In this review, we summarize the progress of research on GacS/GacA from various perspectives, including the reaction mechanism, related regulatory pathways, main functions and GacS/GacA-mediated applications. Hopefully, this review will facilitate further research on GacS/GacA and promote its application in regulating secondary metabolism and as a therapeutic target.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 1","pages":"Article 100051"},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203896","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: 6

Next-generation synthetic biology approaches for the accelerated discovery of microbial natural products 加速发现微生物天然产物的下一代合成生物学方法

Engineering Microbiology Pub Date : 2023-03-01 DOI: 10.1016/j.engmic.2022.100060

Lei Li

{"title":"Next-generation synthetic biology approaches for the accelerated discovery of microbial natural products","authors":"Lei Li","doi":"10.1016/j.engmic.2022.100060","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100060","url":null,"abstract":"<div>Microbial natural products (NPs) and their derivates have been widely used in health care and agriculture during the past few decades. Although large-scale bacterial or fungal (meta)genomic mining has revealed the tremendous biosynthetic potentials to produce novel small molecules, there remains a lack of universal approaches to link NP biosynthetic gene clusters (BGCs) to their associated products at a large scale and speed. In the last ten years, a series of emerging technologies have been established alongside the developments in synthetic biology to engineer cryptic metabolite BGCs and edit host genomes. Diverse computational tools, such as antiSMASH and PRISM, have also been simultaneously developed to rapidly identify BGCs and predict the chemical structures of their products. This review discusses the recent developments and trends pertaining to the accelerated discovery of microbial NPs driven by a wide variety of next-generation synthetic biology approaches, with an emphasis on the in situ activation of silent BGCs at scale, the direct cloning or refactoring of BGCs of interest for heterologous expression, and the synthetic-bioinformatic natural products (syn-BNP) approach for the guided rapid access of bioactive non-ribosomal peptides.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 1","pages":"Article 100060"},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203893","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: 3

Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation 里氏木霉纤维生物水解酶2的组成型过表达揭示了其引发纤维素降解的能力

Engineering Microbiology Pub Date : 2023-03-01 DOI: 10.1016/j.engmic.2022.100059

Yubo Wang , Meibin Ren , Yifan Wang , Lu Wang , Hong Liu , Mei Shi , Yaohua Zhong

{"title":"Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation","authors":"Yubo Wang , Meibin Ren , Yifan Wang , Lu Wang , Hong Liu , Mei Shi , Yaohua Zhong","doi":"10.1016/j.engmic.2022.100059","DOIUrl":"https://doi.org/10.1016/j.engmic.2022.100059","url":null,"abstract":"<div>Cellulose degradation results from the synergistic effect of different enzymes, but which enzyme is involved in the initial stage of cellulose degradation is still not well understood. Cellobiohydrolase 2 (CBH2) attached to the conidial surface is possibly associated with the initial stage. However, its specific mechanism is still incompletely known. This study explored the potential role of CBH2 in initiating cellulose degradation using a constitutive overexpression strategy. First, the CBH2-overexpression Trichoderma reesei strains Qgc2–5 and Qrc2–40 were constructed using the constitutive promoters Pgpd1 and PrpS30, respectively. It was found that cbh2 was expressed at a high level under the glucose conditions and was significantly higher than that of the parental strain QM9414 at the early stage of 29 h when cellulose was used as the carbon source. Particularly, the constitutive overexpression of cbh2 caused the strong expression of major cellulase-encoding genes (cbh1, eg1, and eg2) and the rapid decomposition of cellulosic material. Meanwhile, the scanning electron microscope showed that the groove-like structure of the cellulose surface was eroded seriously owing to CBH2 overexpression, which caused the cellulose surface to be smooth. These results showed that the overexpression of CHB2 caused the major cellulase enzymes to be expressed and contributed to cellulose degradation, showing the potential role of CBH2 in the initial stage of the cellulose hydrolytic process.</div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 1","pages":"Article 100059"},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50203894","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: 3