Synthetic biology aims to establish engineering principles for biological systems by leveraging the design-build-test-learn (DBTL) cycle. Central to the success of the DBTL cycle is the selection of a suitable chassis...
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Synthetic biology aims to establish engineering principles for biological systems by leveraging the design-build-test-learn (DBTL) cycle. Central to the success of the DBTL cycle is the selection of a suitable chassis, which is the environment in which biological designs are tested. Every step of this cycle is strongly influenced by the properties of chassis. A successful chassis must meet various criteria, prompting ongoing research regarding new candidates. Recently, species within the Vibrio genus, notably Vibrio natriegens and related strains, have emerged as promising next-generation chassis due to their rapid growth rates, versatile substrate utilization, and biosafety level 1 classification. These properties make them highly attractive for accelerating the DBTL cycle with the potential for efficient protein and metabolite production. This review emphasizes the foundational requirements for efficient engineering in synthetic biology, including genetic parts, vectors, and genome engineering technologies tailored to Vibrio species. Practical applications, such as metabolic engineering and protein expression, have been discussed, offering a comprehensive summary of recent advances. This paper also outlines the future directions and suggestions for fully unlocking the potential of Vibrio species as next-generation chassis.
BackgroundDNA assembly is an essential technique enabling metabolic engineering and synthetic biology. Combining novel DNA assembly technologies with rational metabolic engineering can facilitate the construction of m...
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BackgroundDNA assembly is an essential technique enabling metabolic engineering and synthetic biology. Combining novel DNA assembly technologies with rational metabolic engineering can facilitate the construction of microbial cell factories. Amino acids and derived biochemicals are important products in industrial biotechnology with wide application and huge markets. DNA assembly scenarios encountered in metabolic engineering for the construction of amino acid and related compound producers, such as design-build-test-learn cycles, construction of precise genetic circuits and repetitive DNA molecules, usually require for iterative, scarless and repetitive sequence assembly methods, *** endonuclease (RE)-assisted strategies constitute one of the major categories of DNA assembly. Here, we developed a Type IIP and IIS RE-assisted method named PS-Brick that comprehensively takes advantage of the properties of PCR fragments and REs for iterative, seamless and repetitive sequence assembly. One round of PS-Brick reaction using purified plasmids and PCR fragments was accomplished within several hours, and transformation of the resultant reaction product from this PS-Brick assembly reaction exhibited high efficiency (10(4)-10(5) CFUs/mu g DNA) and high accuracy (similar to 90%). An application of metabolic engineering to threonine production, including the release of feedback regulation, elimination of metabolic bottlenecks, intensification of threonine export and inactivation of threonine catabolism, was stepwise resolved in E. coli by rounds of design-build-test-learn cycles through the iterative PS-Brick paradigm, and 45.71g/L threonine was obtained through fed-batch fermentation. In addition to the value of the iterative character of PS-Brick for sequential strain engineering, seamless cloning enabled precise in-frame fusion for codon saturation mutagenesis and bicistronic design, and the repetitive sequence cloning ability of PS-Brick enabled
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