Cloning of toyF gene and Its Relevance to Toyocamycin Biosynthesis in Streptomyces diastatochromogenes 1628

doi:10.16409/j.cnki.2095-039x.2016.04.013

Abstract

Abstract: To elucidate the fuction of toyF gene which encoded adenylosuccinate lyase in biosynthesis of toyocamycin (TM) in Streptomyces diastatochromogenes 1628, the toyF gene was successful cloned from S. diastatochromogenes 1628 by using degenerate primer. Then, knock-out plasmid pKC1132-toyF' was constructed and integrated into the chromosome of S. diastatochromogenes 1628 by using intergeneric conjugation to yield strain 1628-ΔTOYF in which the toyF gene was disrupted. Both TOYF activity and TM production of in strain 1628-ΔTOYF decreased 66.7% and 87.5% comparing to those of wild-type strain S. diastatochromogenes 1628, respectively. Moreover, in order to express the toyF gene in strain 1628-ΔTOYF as complementary experiment, recombinant strain 1628-ΔTOYF/toyF was constructed. As a result, the recombinant strain 1628-ΔTOYF/toyF showed an increase of 2.7-fold in specific enzyme activity of TOYF and an increase of 8.3-fold in TM comparing to those of strain 1628-ΔTOYF, respectively. These results suggested that cloned toyF was a key gene which is involved in biosynthesis of TM in S. diastatochromogenes 1628. The cloning and functional analysis of toyF gene will pave the way for cloning the whole gene cluster which required for TM formation. Also, this work provides a basis for studying biosynthetic mechanism of TM in Streptomyces diastatochromogenes 1628.

Key words: Streptomyces diastatochromogenes 1628, toyF gene, gene knockout, toyocamycin, gene cluster

CLC Number:

S476

XU Xianhao, TAO Libing, MA Zheng, YU Xiaoping. Cloning of toyF gene and Its Relevance to Toyocamycin Biosynthesis in Streptomyces diastatochromogenes 1628[J]. journal1, 2016, 32(4): 511-517.

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