論文 - 詳細
| RRC ID | 910 |
|---|---|
| 著者 | Morita Y, Nakamori S, Takagi H. |
| タイトル | L-proline accumulation and freeze tolerance of Saccharomyces cerevisiae are caused by a mutation in the PRO1 gene encoding gamma-glutamyl kinase. |
| ジャーナル | Appl Environ Microbiol |
| Abstract |
We previously isolated a mutant which showed a high tolerance to freezing that correlated with higher levels of intracellular L-proline derived from L-proline analogue-resistant mutants. The mutation responsible for the analogue resistance and L-proline accumulation was a single nuclear dominant mutation. By introducing the mutant-derived genomic library into a non-L-proline-utilizing strain, the mutant was found to carry an allele of the wild-type PRO1 gene encoding gamma-glutamyl kinase, which resulted in a single amino acid replacement; Asp (GAC) at position 154 was replaced by Asn (AAC). Interestingly, the allele of PRO1 was shown to enhance the activities of gamma-glutamyl kinase and gamma-glutamyl phosphate reductase, both of which catalyze the first two steps of L-proline synthesis from L-glutamate and which together may form a complex in vivo. When cultured in liquid minimal medium, yeast cells expressing the mutated gamma-glutamyl kinase were found to accumulate intracellular L-proline and showed a prominent increase in cell viability after freezing at -20 degrees C compared to the viability of cells harboring the wild-type PRO1 gene. These results suggest that the altered gamma-glutamyl kinase results in stabilization of the complex or has an indirect effect on gamma-glutamyl phosphate reductase activity, which leads to an increase in L-proline production in Saccharomyces cerevisiae. The approach described in this paper could be a practical method for breeding novel freeze-tolerant yeast strains. |
| 巻・号 | 69(1) |
| ページ | 212-9 |
| 公開日 | 2003-1-1 |
| DOI | 10.1128/AEM.69.1.212-219.2003 |
| PMID | 12513997 |
| PMC | PMC152471 |
| MeSH | Amino Acid Sequence Amino Acid Substitution* Culture Media Escherichia coli / enzymology Escherichia coli / genetics Freezing Gene Expression Regulation, Fungal Molecular Sequence Data Phosphotransferases (Carboxyl Group Acceptor) / genetics* Phosphotransferases (Carboxyl Group Acceptor) / metabolism Proline / metabolism* Saccharomyces cerevisiae / growth & development* Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae / physiology* Saccharomyces cerevisiae Proteins |
| IF | 4.016 |
| 引用数 | 60 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY |
| リソース情報 | |
| 酵母 | BYP1804(pCgHIS3) |