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Metabolome analysis of metabolic burden in Escherichia coli caused by overexpression of green fluorescent protein and delta-rhodopsin.
Matsuyama, Chinatsu; Seike, Taisuke; Okahashi, Nobuyuki; Niide, Teppei; Hara, Kiyotaka Y; Hirono-Hara, Yoko; Ishii, Jun; Shimizu, Hiroshi; Toya, Yoshihiro; Matsuda, Fumio.
Afiliação
  • Matsuyama C; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan.
  • Seike T; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan.
  • Okahashi N; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan; Osaka University Shimadzu Omics Innovation Research Laboratories, Osaka University, Osaka 565-0871, Japan.
  • Niide T; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan.
  • Hara KY; Department of Environmental and Life Sciences, School of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
  • Hirono-Hara Y; 396Bio Inc., 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
  • Ishii J; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo 657-8501, Japan.
  • Shimizu H; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan.
  • Toya Y; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan.
  • Matsuda F; Graduate School of Information Science and Technology, Osaka University, Osaka 565-0871, Japan; Osaka University Shimadzu Omics Innovation Research Laboratories, Osaka University, Osaka 565-0871, Japan. Electronic address: fmatsuda@ist.osaka-u.ac.jp.
J Biosci Bioeng ; 137(3): 187-194, 2024 Mar.
Article em En | MEDLINE | ID: mdl-38281859
ABSTRACT
Overexpression of proteins by introducing a DNA vector is among the most important tools for the metabolic engineering of microorganisms such as Escherichia coli. Protein overexpression imposes a burden on metabolism because metabolic pathways must supply building blocks for protein and DNA synthesis. Different E. coli strains have distinct metabolic capacities. In this study, two proteins were overexpressed in four E. coli strains (MG1655(DE3), W3110(DE3), BL21star(DE3), and Rosetta(DE3)), and their effects on metabolic burden were investigated. Metabolomic analysis showed that E. coli strains overexpressing green fluorescent protein had decreased levels of several metabolites, with a positive correlation between the number of reduced metabolites and green fluorescent protein expression levels. Moreover, nucleic acid-related metabolites decreased, indicating a metabolic burden in the E. coli strains, and the growth rate and protein expression levels were improved by supplementation with the five nucleosides. In contrast, two strains overexpressing delta rhodopsin, a microbial membrane rhodopsin from Haloterrigena turkmenica, led to a metabolic burden and decrease in the amino acids Ala, Val, Leu, Ile, Thr, Phe, Asp, and Trp, which are the most frequent amino acids in the delta rhodopsin protein sequence. The metabolic burden caused by protein overexpression was influenced by the metabolic capacity of the host strains and the sequences of the overexpressed proteins. Detailed characterization of the effects of protein expression on the metabolic state of engineered cells using metabolomics will provide insights into improving the production of target compounds.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Rodopsina / Escherichia coli Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Rodopsina / Escherichia coli Idioma: En Ano de publicação: 2024 Tipo de documento: Article