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KMID : 0545120120220060793
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Journal of Microbiology and Biotechnology
2012 Volume.22 No. 6 p.793 ~ p.799
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Isolation and Functional Analysis of spy1 Responsible for Pristinamycin Yield in Streptomyces pristinaespiralis
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Qingchao Jin
Huali Yin
Xiaowei Hong
Zhihua Jin
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Abstract
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A gene related to high pristinamycin yield in Streptomyces pristinaespiralis was selected by amplified fragment length polymorphism (AFLP) and its functions were investigated by gene disruption. First, a 561 bp polymorphic sequence was acquired by AFLP from high-yield recombinants compared with the S. pristinaespiralis ancestor ATCC25486, indicating that this approach is an effective means of screening for valuable genes responsible for antibiotic yield. Then, a 2,127 bp open reading frame of a gene designated spy1 that overlaps with the above fragment was identified and its structure and biological functions were investigated. In silico analysis of spy1 encoding a deduced 708-amino-acid-long serine/threonine protein kinase showed that it only contains a catalytic domain in the N-terminal region, which is different from some known homologs. Gene inactivation of chromosomal spy1 indicated that it plays a pleiotropic regulatory function in pristinamycin production, with a positive correlation to pristinamycin I biosynthesis and a negative correlation to pristinamycin II biosynthesis.
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KEYWORD
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Streptomyces pristinaespiralis, pristinamycin, yield, gene screening, gene disruption
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