| RRC ID |
50810
|
| 著者 |
Kasuga K, Sasaki A, Matsuo T, Yamamoto C, Minato Y, Kuwahara N, Fujii C, Kobayashi M, Agematu H, Tamura T, Komatsu M, Ishikawa J, Ikeda H, Kojima I.
|
| タイトル |
Heterologous production of kasugamycin, an aminoglycoside antibiotic from Streptomyces kasugaensis, in Streptomyces lividans and Rhodococcus erythropolis L-88 by constitutive expression of the biosynthetic gene cluster.
|
| ジャーナル |
Appl Microbiol Biotechnol
|
| Abstract |
Kasugamycin (KSM), an aminoglycoside antibiotic isolated from Streptomyces kasugaensis cultures, has been used against rice blast disease for more than 50 years. We cloned the KSM biosynthetic gene (KBG) cluster from S. kasugaensis MB273-C4 and constructed three KBG cassettes (i.e., cassettes I-III) to enable heterologous production of KSM in many actinomycetes by constitutive expression of KBGs. Cassette I comprised all putative transcriptional units in the cluster, but it was placed under the control of the P neo promoter from Tn5. It was not maintained stably in Streptomyces lividans and did not transform Rhodococcus erythropolis. Cassette II retained the original arrangement of KBGs, except that the promoter of kasT, the specific activator gene for KBG, was replaced with P rpsJ , the constitutive promoter of rpsJ from Streptomyces avermitilis. To enhance the intracellular concentration of myo-inositol, an expression cassette of ino1 encoding the inositol-1-phosphate synthase from S. avermitilis was inserted into cassette II to generate cassette III. These two cassettes showed stable maintenance in S. lividans and R. erythropolis to produce KSM. Particularly, the transformants of S. lividans induced KSM production up to the same levels as those produced by S. kasugaensis. Furthermore, cassette III induced more KSM accumulation than cassette II in R. erythropolis, suggesting an exogenous supply of myo-inositol by the ino1 expression in the host. Cassettes II and III appear to be useful for heterologous KSM production in actinomycetes. Rhodococcus exhibiting a spherical form in liquid cultivation is also a promising heterologous host for antibiotic fermentation.
|
| 巻・号 |
101(10)
|
| ページ |
4259-4268
|
| 公開日 |
2017-5-1
|
| DOI |
10.1007/s00253-017-8189-5
|
| PII |
10.1007/s00253-017-8189-5
|
| PMID |
28243709
|
| MeSH |
Aminoglycosides / biosynthesis*
Anti-Bacterial Agents / biosynthesis*
Base Sequence
Cloning, Molecular
Fermentation
Gene Expression Regulation, Bacterial
Genes, Bacterial
Inositol / biosynthesis
Inositol / metabolism
Multigene Family*
Myo-Inositol-1-Phosphate Synthase / genetics
Myo-Inositol-1-Phosphate Synthase / metabolism
Rhodococcus / genetics*
Rhodococcus / metabolism
Streptomyces / genetics*
Streptomyces / metabolism
Streptomyces lividans / genetics*
Transcription Factors / metabolism
|
| IF |
3.53
|
| 引用数 |
8
|
| リソース情報 |
| 一般微生物 |
JCM 3201 |
