Toyocamycin exhibits effective biological activities for use against plant pathogenic fungi thanks to its structural similarity to nucleoside. It has been recognized as a promising agricultural antibiotic utilized in controlling the occurrence of plant diseases. In our previous study, a strain that was isolated was identified and designated as Streptomyces diastatochromogenes whose major secondary metabolite was toyocamycin, but the production was largely limited. Protoplast transformation is a useful technique in the improvement of streptomycete. In this study, we optimized some key factors necessary for protoplast formation, regeneration, and transformation of S. diastatochromogenes. When mycelium was cultivated in CP medium with 1 % glycine, harvested at 48 h old, and then treated with 3 mg lysozyme/mL in P buffer for 1 h, the greatest regeneration frequency (42.5 %) of protoplasts was obtained. By using 1 × 10(9)/mL protoplasts with polyethylene glycol 1000 at a concentration of 30 % (w/v), the best performance of protoplast transformation efficiency was 4.8 × 10(3)/μg DNA transformants.

Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine College of Life Sciences China Jiliang University Xueyuan Street Xiasha Higher Education District Hangzhou Zhejiang Province 310018 China

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Enzyme Microb Technol. 2011 Jun 10;49(1):17-24 PubMed

Nature. 1978 Jul 27;274(5669):398-400 PubMed

Curr Microbiol. 2005 Aug;51(2):116-21 PubMed

Org Biomol Chem. 2011 Apr 7;9(7):2227-32 PubMed

J Antibiot (Tokyo). 1988 Dec;41(12):1711-39 PubMed

Metab Eng. 2008 Sep;10(5):281-92 PubMed

Appl Microbiol Biotechnol. 1990 Jan;32(4):431-5 PubMed

J Gen Microbiol. 1989 Aug;135(8):2289-97 PubMed

Antibiot Khimioter. 1990 Jul;35(7):3-5 PubMed

Antibiot Khimioter. 2000;45(4):10-3 PubMed

Curr Microbiol. 2004 Dec;49(6):400-6 PubMed

Lett Appl Microbiol. 1993 Apr;16(4):217-9 PubMed

J Bacteriol. 1985 Jul;163(1):180-5 PubMed

Antibiot Khimioter. 1988 Mar;33(3):180-5 PubMed