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Adzemye Fovennso Bridget,Rubin Thapa Magar,Jae Kyung Sohng 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
Saccharapolyspora spinosa is the producer of the environmentally friendly insecticide spinosad, which is a combination of spinosyn A and D. spinosyn is a tetracyclic polyketide aglycone to which is attached a neutral saccharide substituent (2,3,4-O-trimethyla-L-rhamnosyl) on the C-9 hydroxyl group and an aminosugar moiety (β D-forosaminyl) on the C-17 hydroxyl group). Both the rhamnose and forosamine moieties are essential for the insecticidal activity of spinosyns. We construct a vector; pIBR-SPN, cloned the sugar genes to it and transformed to Sa. spinosa. The best strain Sa. spinosa MUV pIBR-SPN FR produced 1,896.77 mg/L of spinosad, 17 folds higher than parent strain.
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Identification of 1,3,6,8- Tetrahydroxynaphthalene Synthase (ThnA) from Nocardia sp. CS682
( Purna Bahadur Poudel ),( Rubin Thapa Magar ),( Adzemye Fovennso Bridget ),( Jae Kyung Sohng ) 한국미생물생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.7
Type III polyketide synthase (PKS) found in bacteria is known as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Microbial type III PKSs synthesize various compounds that possess crucial biological functions and significant pharmaceutical activities. Based on our sequence analysis, we have identified a putative type III polyketide synthase from Nocardia sp. CS682 was named as ThnA. The role of ThnA, in Nocardia sp. CS682 during the biosynthesis of 1,3,6,8 tetrahydroxynaphthalene (THN), which is the key intermediate of 1-(α-L-(2-O-methyl)-6-deoxymannopyranosyloxy)-3,6,8- trimethoxynaphthalene (IBR-3) was characterized. ThnA utilized five molecules of malonyl-CoA as a starter substrate to generate the polyketide 1,3,6,8-tetrahydroxynaphthalene, which could spontaneously be oxidized to the red flaviolin compound 2,5,7-trihydroxy-1,4-naphthoquinone. The amino acid sequence alignment of ThnA revealed similarities with a previously identified type III PKS and identified Cys<sup>138</sup>, Phe<sup>188</sup>, His<sup>270</sup>, and Asn<sup>303</sup> as four highly conserved active site amino acid residues, as found in other known polyketide synthases. In this study, we report the heterologous expression of the type III polyketide synthase thnA in S. lividans TK24 and the identification of THN production in a mutant strain. We also compared the transcription level of thnA in S. lividans TK24 and S. lividans pIBR25-thnA and found that thnA was only transcribed in the mutant.
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