Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae

Tang, Hongting; Wu, Yanling; Deng, Jiliang; Chen, Nanzhu; Zheng, Zhaohui; Wei, Yongjun; Luo, Xiaozhou; Keasling, Jay D.

doi:10.3390/metabo10080320

Cited by 61 publications

(47 citation statements)

References 153 publications

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“…For example, the T7 promoter has high strength and is most commonly used for driving gene expression in E. coli (Hawley & McClure, 1983 ). Indeed, the discovery, engineering and characterization of effective promoters have been the focus of metabolic engineering research for overproducing NPs in different microbes, which has been well-reviewed elsewhere (Blazeck & Alper, 2013 ; Jin et al., 2019 ; Tang et al., 2020 ).…”

Section: Metabolic Engineering and Synthetic Biology Toolkits For Heterologous Production Of Cyanobacterial Compoundsmentioning

confidence: 99%

Heterologous production of cyanobacterial compounds

Dhakal

Chen

Luesch

et al. 2021

Journal of Industrial Microbiology and Biotechnology

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Cyanobacteria produce a plethora of compounds with unique chemical structures and diverse biological activities. Importantly, the increasing availability of cyanobacterial genome sequences and the rapid development of bioinformatics tools have unraveled the tremendous potential of cyanobacteria in producing new natural products. However, the discovery of these compounds based on cyanobacterial genomes has progressed slowly as the majority of their corresponding biosynthetic gene clusters (BGCs) are silent. In addition, cyanobacterial strains are often slow-growing, difficult for genetic engineering, or cannot be cultivated yet, limiting the use of host genetic engineering approaches for discovery. On the other hand, genetically tractable hosts such as Escherichia coli, Actinobacteria, and yeast have been developed for the heterologous expression of cyanobacterial BGCs. More recently, there has been increased interests in developing model cyanobacterial strains as heterologous production platforms. Herein, we present recent advances in the heterologous production of cyanobacterial compounds in both cyanobacterial and non-cyanobacterial hosts. Emerging strategies for BGC assembly, host engineering, and optimization of BGC expression are included for fostering the broader applications of synthetic biology tools in the discovery of new cyanobacterial natural products.

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Section: Metabolic Engineering and Synthetic Biology Toolkits For Heterologous Production Of Cyanobacterial Compoundsmentioning

confidence: 99%

Heterologous production of cyanobacterial compounds

Dhakal

Chen

Luesch

et al. 2021

Journal of Industrial Microbiology and Biotechnology

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“…The artificially constructed cell factories need to be continuously optimized through the Design-Build-Test-Learn (DBTL) cycle (Nielsen and Keasling, 2016). The DBTL cycle includes new enzyme discovery, heterologous gene expression, promoter engineering, metabolic flux balance, pathway optimization, oxidation and reduction system balance, genome-scale metabolic models and other metabolic engineering strategies (Jiang et al, 2020;Ko et al, 2020;Tang et al, 2020;Wang M. et al, 2020). Finally, a high-yield GA or other natural product microbial cell factory can be obtained.…”

Section: Metabolic Engineering Of S Cerevisiae For Ga Productionmentioning

confidence: 99%

Metabolic Engineering for Glycyrrhetinic Acid Production in Saccharomyces cerevisiae

Guan

Wang

Guan

et al. 2020

Front. Bioeng. Biotechnol.

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Glycyrrhetinic acid (GA) is one of the main bioactive components of licorice, and it is widely used in traditional Chinese medicine due to its hepatoprotective, immunomodulatory, anti-inflammatory and anti-viral functions. Currently, GA is mainly extracted from the roots of cultivated licorice. However, licorice only contains low amounts of GA, and the amount of licorice that can be planted is limited. GA supplies are therefore limited and cannot meet the demands of growing markets. GA has a complex chemical structure, and its chemical synthesis is difficult, therefore, new strategies to produce large amounts of GA are needed. The development of metabolic engineering and emerging synthetic biology provide the opportunity to produce GA using microbial cell factories. In this review, current advances in the metabolic engineering of Saccharomyces cerevisiae for GA biosynthesis and various metabolic engineering strategies that can improve GA production are summarized. Furthermore, the advances and challenges of yeast GA production are also discussed. In summary, GA biosynthesis using metabolically engineered S. cerevisiae serves as one possible strategy for sustainable GA supply and reasonable use of traditional Chinese medical plants.

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“…However, not many native promoters have been characterized deeply, which implies a limited dynamic range for gene expression within synthetic circuits. To overcome this problem, libraries of synthetic promoters of different strengths have been developed using error-prone PCR on a single template [ 18 ] or building hybrid promoters by combining UASs and core sequences from different yeast promoters [ 19 , 20 ]. These synthetic promoters, however, are not orthogonal to S. cerevisiae cells and could also undergo homologous recombination with their original copy in the yeast genome.…”

Section: Introductionmentioning

confidence: 99%

Novel S. cerevisiae Hybrid Synthetic Promoters Based on Foreign Core Promoter Sequences

Feng

Marchisio

2021

IJMS

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Promoters are fundamental components of synthetic gene circuits. They are DNA segments where transcription initiation takes place. New constitutive and regulated promoters are constantly engineered in order to meet the requirements for protein and RNA expression into different genetic networks. In this work, we constructed and optimized new synthetic constitutive promoters for the yeast Saccharomyces cerevisiae. We started from foreign (e.g., viral) core promoters as templates. They are, usually, unfunctional in yeast but can be activated by extending them with a short sequence, from the CYC1 promoter, containing various transcription start sites (TSSs). Transcription was modulated by mutating the TATA box composition and varying its distance from the TSS. We found that gene expression is maximized when the TATA box has the form TATAAAA or TATATAA and lies between 30 and 70 nucleotides upstream of the TSS. Core promoters were turned into stronger promoters via the addition of a short UAS. In particular, the 40 nt bipartite UAS from the GPD promoter can enhance protein synthesis considerably when placed 150 nt upstream of the TATA box. Overall, we extended the pool of S. cerevisiae promoters with 59 new samples, the strongest overcoming the native TEF2 promoter.

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Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae

Cited by 61 publications

References 153 publications

Heterologous production of cyanobacterial compounds

Heterologous production of cyanobacterial compounds

Metabolic Engineering for Glycyrrhetinic Acid Production in Saccharomyces cerevisiae

Novel S. cerevisiae Hybrid Synthetic Promoters Based on Foreign Core Promoter Sequences

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