Identification and engineering of regulation-related genes toward improved kasugamycin production
Identification and engineering of regulation-related genes toward improved kasugamycin production
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调控相关基因的鉴定和改造以提高春雷霉素的产量
DOI：
10.1007/s00253-015-7082-3
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发表时间：
2016-02
影响因子：
5
通讯作者：
Deng, Zixin
中科院分区：
文献类型：
--
作者：
Kang, Qianjin;Bai, Linquan;Cheng, Lin;Deng, Zixin
春日霉素由春日链霉菌和小金黄色葡萄球菌产生，是一种重要的氨基糖苷类抗生素，广泛用于兽医和农业应用。在先前报道的春雷霉素基因簇的左翼区域中，在低产S中鉴定出了四个额外的基因（双组分系统kasWandkasX、MerR家族kasV和异戊二烯基半胱氨酸羧基甲基转移酶kasS）。 kasugaensisBCRC12349 和高产 S.微金黄色葡萄球菌XM301。调控基因的缺失会导致春雷霉素的产生停止，并且其在 BCRC12349 中的过度表达导致滴度增加 186%。删除 kasW、kasX、kasV 和 kas 分别将春雷霉素产量提高 12%、19%、194% 和 22%。 qRT-PCR 分析表明，所有四种突变体中 kasgenes 的转录均显着增加。在高产菌株S中也进行了类似的基因失活。微金黄色葡萄球菌XM301。正如预期的那样，删除 kasW/X 导致产量增加 58%，从 6 g/L 增加到 9.5 g/L。然而，kasV的缺失和kasT的过表达没有明显的影响，并且kasS的破坏令人惊讶地降低了春雷霉素的产量。此外，thekasSmutant 与 TTA 密码子突变的 kasS 的反式互补使春雷霉素产量增加了 20%。在高产 XM301 中检测到的 kasgenes 转录比在低产 BCRC12349 中高得多，这可能部分解释了它们之间基因失活效果的差异。我们的工作不仅产生了春雷霉素产量提高的工程菌株，而且还指出对于低产或高产菌株应考虑不同的调控相关基因的操作策略。
Kasugamycin, produced byStreptomyces kasugaensisandStreptomyces microaureus, is an important amino-glycoside family antibiotic and widely used for veterinary and agricultural applications. In the left flanking region of the previously reported kasugamycin gene cluster, four additional genes (two-component systemkasWandkasX, MerR-familykasV, and isoprenylcysteine carboxyl methyltransferasekasS) were identified both in the low-yieldingS. kasugaensisBCRC12349 and high-yieldingS. microaureusXM301. Deletion of regulatory genekasTabolished kasugamycin production, and its overexpression in BCRC12349 resulted in an increased titer by 186 %. Deletion ofkasW,kasX,kasV, andkasSimproved kasugamycin production by 12, 19, 194, and 22 %, respectively. qRT-PCR analysis demonstrated that the transcription ofkasgenes was significantly increased in all the four mutants. Similar gene inactivation was performed in the high-yielding strainS. microaureusXM301. As expected, the deletion ofkasW/Xresulted in a 58 % increase of the yield from 6 to 9.5 g/L. However, the deletion ofkasVand over-expression ofkasThad no obvious effect, and the disruption ofkasSsurprisingly decreased kasugamycin production. In addition, trans-complementation of thekasSmutant with a TTA codon-mutatedkasSincreased the kasugamycin yield by 20 %. A much higher transcription ofkasgenes was detected in the high-yielding XM301 than in the low-yielding BCRC12349, which may partially account for the discrepancy of gene inactivation effects between them. Our work not only generated engineered strains with improved kasugamycin yield, but also pointed out that different strategies on manipulating regulatory-related genes should be considered for low-yielding or high-yielding strains.
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影响因子：
2.1
作者：
Li Dong;N. Nakashima;N. Tamura;T. Tamura
通讯作者：
Li Dong;N. Nakashima;N. Tamura;T. Tamura
DOI：
10.7164/antibiotics.55.1053
发表时间：
2002-12
期刊：
The Journal of antibiotics
影响因子：
--
作者：
S. Ikeno;D. Aoki;Koji Sato;M. Hamada;M. Hori;K. Tsuchiya
通讯作者：
S. Ikeno;D. Aoki;Koji Sato;M. Hamada;M. Hori;K. Tsuchiya
影响因子：
1.2
作者：
C. Wilkinson;Z. Hughes-Thomas;Christine J. Martin;I. Böhm;T. Mironenko;M. Deacon;M. Wheatcroft;G. Wirtz;J. Staunton;P. Leadlay
通讯作者：
C. Wilkinson;Z. Hughes-Thomas;Christine J. Martin;I. Böhm;T. Mironenko;M. Deacon;M. Wheatcroft;G. Wirtz;J. Staunton;P. Leadlay
影响因子：
3.3
作者：
Ikeno, S;Tsuji, T;Hori, M
通讯作者：
Hori, M
影响因子：
5
作者：
Zhao, Youbao;Xiang, Sihai;Dai, Xida;Yang, Keqian
通讯作者：
Yang, Keqian