Improvement of Natamycin Production by Engineering of Phosphopantetheinyl Transferases in Streptomyces chattanoogensis L10
Improvement of Natamycin Production by Engineering of Phosphopantetheinyl Transferases in Streptomyces chattanoogensis L10
复制标题
通过改造链霉菌 Chattanoogensis L10 中的磷酸泛酰乙胺基转移酶来提高那他霉素的生产
DOI：
10.1128/aem.00099-13
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发表时间：
2013-06-01
影响因子：
4.4
通讯作者：
Li, Yong-Quan
中科院分区：
文献类型：
--
作者：
Jiang, Hui;Wang, Yue-Yue;Li, Yong-Quan
摘要磷酸泛酰巯基乙胺基转移酶（PPTases）通过将I/II型聚酮合酶（PKS）和非核糖体肽合成酶（NRPS）中的酰基载体蛋白（ACP）和NRPS中的肽基载体蛋白（PCP）从无活性的脱辅基形式转化为有活性的完整形式，从而导致聚酮和非核糖体肽的生物合成，对I/II型聚酮合酶（PKS）和非核糖体肽合成酶（NRPS）的活性至关重要。工业纳他霉素（NTM）生产商，查塔努加链霉菌L10，含有两个PPT酶（SchPPT和SchACPS）和五个PKS。这两种PPT酶的生化特性表明，SchPPT催化I型和II型PKS中ACP的磷酸泛酰巯基乙胺化，SchACPS催化II型PKS和脂肪酸脱氢酶（FAS）中ACP的磷酸泛酰巯基乙胺化，SchPPT的特异性可能受其C末端控制。SchPPT在S. Chattanoogensis L10抑制了NTM的产生，但没有抑制孢子色素的产生，而SchPPT基因的过表达不仅使NTM的产生增加了约40%，而且还加速了NTM和孢子色素的产生。因此，我们阐明了一个全面的磷酸泛酰巯基乙胺化网络的PKS和改进的聚酮化合物的生产，通过工程化的同源PPT酶在细菌中。
ABSTRACT Phosphopantetheinyl transferases (PPTases) are essential to the activities of type I/II polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) through converting acyl carrier proteins (ACPs) in PKSs and peptidyl carrier proteins (PCPs) in NRPSs from inactive apo-forms into active holo-forms, leading to biosynthesis of polyketides and nonribosomal peptides. The industrial natamycin (NTM) producer, Streptomyces chattanoogensis L10, contains two PPTases (SchPPT and SchACPS) and five PKSs. Biochemical characterization of these two PPTases shows that SchPPT catalyzes the phosphopantetheinylation of ACPs in both type I PKSs and type II PKSs, SchACPS catalyzes the phosphopantetheinylation of ACPs in type II PKSs and fatty acid synthases (FASs), and the specificity of SchPPT is possibly controlled by its C terminus. Inactivation of SchPPT in S. chattanoogensis L10 abolished production of NTM but not the spore pigment, while overexpression of the SchPPT gene not only increased NTM production by about 40% but also accelerated productions of both NTM and the spore pigment. Thus, we elucidated a comprehensive phosphopantetheinylation network of PKSs and improved polyketide production by engineering the cognate PPTase in bacteria.