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d ‐amino acid auxotrophic Escherichia coli strain for in vivo functional cloning of novel d ‐amino acid synthetic enzyme by Tomokazu Ito; Natsumi Muto; Haruna Sakagami; Miho Tanaka; Hisashi Hemmi; Tohru Yoshimura is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is d ‐amino acid auxotrophic Escherichia coli strain for in vivo functional cloning of novel d ‐amino acid synthetic enzyme about?
Various d -amino acids have been found in a wide range of organisms, including mammals. Although the physiological functions of various d -amino acids have been reported or suggested, the molecular basis of these biological functions has been elucidated in only a few cases. The identification of a d -amino acid biosynthetic enzyme is a critical step in understanding the mechanism of the physiological functions of d -amino acids. While in vivo functional screening can be a powerful tool for identifying novel metabolic enzymes, none of the existing organisms exhibit growth dependent on d -amino acid other than d -Ala and d -Glu. Here, we report the first organism that exhibits non-canonical d -amino acid auxotrophy. We found that an Escherichia coli strain lacking the major d -Ala and d -Glu biosynthetic enzymes, alr , dadX , and murI , and expressing the mutated d -amino acid transaminase (DAAT) gene from Bacillus sp. YM-1 (MB3000/m daat + ) grew well when supplemented with certain d -amino acid. A multicopy suppression study with plasmids encoding one of the 51 PLP-dependent enzymes of E. coli showed that MB3000/m daat + could detect weak and moonlighting racemase activity, such fr
Who reads d ‐amino acid auxotrophic Escherichia coli strain for in vivo functional cloning of novel d ‐amino acid synthetic enzyme?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Tomokazu Ito; Natsumi Muto; Haruna Sakagami; Miho Tanaka; Hisashi Hemmi; Tohru Yoshimura
- Publisher
- Wiley
- Published
- 2023
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)