High-level production of ornithine by expression of the feedback inhibition-insensitive N-acetyl glutamate kinase in the sake yeast Saccharomyces cerevisiae

• Ohashi, Masataka

• Nasuno, Ryo

• Isogai, Shota

• Takagi, Hiroshi

Bibliographic Information

Other Title

• 清酒酵母におけるフィードバック阻害非感受性型N-アセチルグルタミン酸キナーゼの発現によるオルニチン高生産

Abstract

We previously reported that intracellular proline (Pro) confers tolerance to ethanol on the yeast Saccharomyces cerevisiae. In this study, to improve the ethanol productivity of sake, a traditional Japanese alcoholic beverage, we successfully isolated several Pro-accumulating mutants derived from diploid sake yeast of S. cerevisiae by a conventional mutagenesis. Interestingly, one of them (strain A902-4) produced more than 10-fold greater amounts of ornithine (Orn) and Pro compared to the parent strain (K901). Orn is a non-proteinogenic amino acid and a precursor of both arginine (Arg) and Pro. It has some physiological functions, such as amelioration of negative states such as lassitude and improvement of sleep quality. We also identified a homo-allelic mutation in the ARG5,6 gene encoding the Thr340Ile variant N-acetylglutamate kinase (NAGK) in strain A902-4. The NAGK activity of the Thr340Ile variant was extremely insensitive to feedback inhibition by Arg, leading to intracellular Orn accumulation. This is the first report of the removal of feedback inhibition of NAGK activity in the industrial yeast, leading to high levels of intracellular Orn. Moreover, sake and sake cake brewed with strain A902-4 contained 4?5 times more Orn than those brewed with strain K901. The approach described here could be a practical method for the development of industrial yeast strains with overproduction of Orn.

Journal

• Metabolic Engineering

  Metabolic Engineering 62 1-9, 2020-11-01

  Elsevier

Keywords

• Saccharomyces cerevisiae
• Sake yeast
• Ornithine
• N-acetyl glutamate kinase Arg5,6
• Feedback inhibition

Details

• CRID: 1050577309352751360
• NII Article ID: 120007147753
• ISSN: 10967176
• DOI: 10.1016/j.ymben.2020.08.005
• HANDLE: 10061/14447
• Web Site: https://naist.repo.nii.ac.jp/records/9351
• Web Site: https://api.elsevier.com/content/article/PII:S1096717620301257?httpAccept=text/xml
• Web Site: https://api.elsevier.com/content/article/PII:S1096717620301257?httpAccept=text/plain
• Text Lang: en
• Article Type: journal article
• Data Source:
  • IRDB
  • Crossref
  • CiNii Articles
  • KAKEN