Roberto Giuseppe Stella , Philipp Baumann , Sophia Lorke , Felix Münstermann , Astrid Wirtz , Johanna Wiechert , Jan Marienhagen , Julia Frunzke
{"title":"Biosensor-based isolation of amino acid-producing Vibrio natriegens strains","authors":"Roberto Giuseppe Stella , Philipp Baumann , Sophia Lorke , Felix Münstermann , Astrid Wirtz , Johanna Wiechert , Jan Marienhagen , Julia Frunzke","doi":"10.1016/j.mec.2021.e00187","DOIUrl":null,"url":null,"abstract":"<div><p>The marine bacterium <em>Vibrio natriegens</em> has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. <em>V. natriegens</em> is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of under 10 min. However, <em>V. natriegens</em> is not an established model organism yet, and further research is required to promote its transformation into a microbial workhorse.</p><p>In this work, the potential of <em>V. natriegens</em> as an amino acid producer was investigated. First, the transcription factor-based biosensor LysG, from <em>Corynebacterium glutamicum</em>, was adapted for expression in <em>V. natriegens</em> to facilitate the detection of positively charged amino acids. A set of different biosensor variants were constructed and characterized, using the expression of a fluorescent protein as sensor output. After random mutagenesis, one of the LysG-based sensors was used to screen for amino acid producer strains. Here, fluorescence-activated cell sorting enabled the selective sorting of highly fluorescent cells, <em>i.e.</em> potential producer cells. Using this approach, individual L-lysine, L-arginine and L-histidine producers could be obtained producing up to 1 mM of the effector amino acid, extracellularly. Genome sequencing of the producer strains provided insight into the amino acid production metabolism of <em>V. natriegens</em>.</p><p>This work demonstrates the successful expression and application of transcription factor-based biosensors in <em>V. natriegens</em> and provides insight into the underlying physiology, forming a solid basis for further development of this promising microbe.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":"13 ","pages":"Article e00187"},"PeriodicalIF":3.7000,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ca/05/main.PMC8605253.pdf","citationCount":"5","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolic Engineering Communications","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214030121000274","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 5
Abstract
The marine bacterium Vibrio natriegens has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. V. natriegens is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of under 10 min. However, V. natriegens is not an established model organism yet, and further research is required to promote its transformation into a microbial workhorse.
In this work, the potential of V. natriegens as an amino acid producer was investigated. First, the transcription factor-based biosensor LysG, from Corynebacterium glutamicum, was adapted for expression in V. natriegens to facilitate the detection of positively charged amino acids. A set of different biosensor variants were constructed and characterized, using the expression of a fluorescent protein as sensor output. After random mutagenesis, one of the LysG-based sensors was used to screen for amino acid producer strains. Here, fluorescence-activated cell sorting enabled the selective sorting of highly fluorescent cells, i.e. potential producer cells. Using this approach, individual L-lysine, L-arginine and L-histidine producers could be obtained producing up to 1 mM of the effector amino acid, extracellularly. Genome sequencing of the producer strains provided insight into the amino acid production metabolism of V. natriegens.
This work demonstrates the successful expression and application of transcription factor-based biosensors in V. natriegens and provides insight into the underlying physiology, forming a solid basis for further development of this promising microbe.
期刊介绍:
Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.