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KMID : 0545120030130050819
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 5 p.819 ~ p.822
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Heterologous Expression of Hybrid Type Ⅱ Polyketide Synthase System in Streptomyces Species
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KIM, CHANG-YOUNG
PARK, HYUN-JOO/KIM,EUNG-SOO
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Abstract
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Polyketides are an extensive class of secondary metabolites with diverse molecular structures and biological activities. A plasmid-based minimal polyketide synthase (PKS) expression cassette was constructed using a subset of actinorhodin (act) biosynthetic genes ( act1 - orfl , ac1-orj2, actl-orjj´, actlll, actⅦ, and actⅣ) from Streptomyces coelicolor, which specify the construction of an orange-fluorescent anthraquinone product aloesaponarin 11, a type Ⅱ polyketide compound derived from one acetyl coenzyme A and 7 malonyl coenzyme A extender units. This system was designed as an indicator pathway in S. pavulus to generate a hybrid type I1 polyketide compound via gene-specific replacement. The act P-ketoacyl synthase unit (actl-orf1 and actl-orf2) in the expression cassette was specifically replaced with oxytetracycline β-ketoacyl synthase (otcY-orf1 and otcY-orf2). This plasmid-based hybrid PKS cassette generated a novel orange-fluorescent compound structurally different from aloesaponarin I1 in both S. lividans and S. arvulus. In addition, several additional distinctive blue-fluorescent compounds were detected, when this hybrid PKS cassette was expressed in S. coelicolor B78 (actl-orf2 mutant), implying that the expression of plasmid-based hybrid PKS cassette in Streptomyces species should be an efficient way of generating hybrid type I1 polyketide compounds.
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