Streptomyces lividans Blasticidin S Deaminase and Its Application in Engineering a Blasticidin S-Producing Strain for Ease of Genetic Manipulation
Streptomyces lividans Blasticidin S Deaminase and Its Application in Engineering a Blasticidin S-Producing Strain for Ease of Genetic Manipulation
复制标题
浅青链霉菌杀稻瘟菌素 S 脱氨酶及其在工程化杀稻瘟菌素 S 生产菌株以方便基因操作中的应用
DOI：
10.1128/aem.03254-12
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发表时间：
2013-04-01
影响因子：
4.4
通讯作者：
He, Xinyi
中科院分区：
文献类型：
--
作者：
Li, Li;Wu, Jun;He, Xinyi
摘要杀稻瘟菌素S是由灰色产色链霉菌产生的一种肽基核苷类抗生素，具有很强的杀菌活性。为了克服天然生产菌的DNA吸收屏障系统，并研究其在体内的生物合成，将杀稻瘟菌素S生物合成基因簇（bls）嫁接到变铅青链霉菌的染色体上。然而，所得突变体LL 2产生无活性的脱氨基羟基杀稻瘟菌素S而不是杀稻瘟菌素S。随后，杀稻瘟菌素S脱氨酶（SLBSD，用于S.在S.并显示控制这种体内转化。发现纯化的SLBSD能够在体外将杀稻瘟菌素S转化为脱氨基羟基杀稻瘟菌素S。它还催化胞苷葡萄糖醛酸（杀稻瘟菌素S生物合成的中间体）的胞嘧啶部分的脱氨基作用。在S.在所得到的突变体S.变铅青WJ 2.为了证明杀稻瘟菌素S生物合成基因簇，blsE，blsF和blsL，编码预测的自由基S-腺苷甲硫氨酸（SAM）蛋白，未知的蛋白质，和胍基甲基转移酶的容易操作，分别失活，以访问它们在杀稻瘟菌素S生物合成中的作用。
ABSTRACT Blasticidin S is a peptidyl nucleoside antibiotic produced by Streptomyces griseochromogenes that exhibits strong fungicidal activity. To circumvent an effective DNA uptake barrier system in the native producer and investigate its biosynthesis in vivo, the blasticidin S biosynthetic gene cluster (bls) was engrafted to the chromosome of Streptomyces lividans. However, the resulting mutant, LL2, produced the inactive deaminohydroxyblasticidin S instead of blasticidin S. Subsequently, a blasticidin S deaminase (SLBSD, for S. lividans blasticidin S deaminase) was identified in S. lividans and shown to govern this in vivo conversion. Purified SLBSD was found to be capable of transforming blasticidin S to deaminohydroxyblasticidin S in vitro. It also catalyzed deamination of the cytosine moiety of cytosylglucuronic acid, an intermediate in blasticidin S biosynthesis. Disruption of the SLBSD gene in S. lividans LL2 led to successful production of active blasticidin S in the resultant mutant, S. lividans WJ2. To demonstrate the easy manipulation of the blasticidin S biosynthetic gene cluster, blsE, blsF, and blsL, encoding a predicted radical S-adenosylmethionine (SAM) protein, an unknown protein, and a guanidino methyltransferase, were individually inactivated to access their role in blasticidin S biosynthesis.