Huang Kai, Zhang Bo, Chen Yu, Liu Zhi-Qiang, Zheng Yu-Guo
The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, China.
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China.
Front Bioeng Biotechnol. 2021 Feb 1;8:621431. doi: 10.3389/fbioe.2020.621431. eCollection 2020.
Antibiotics play an important role in human health. Most antibiotics are derived from microbial secondary metabolites. Amphotericin is a polyene macrolide antibiotic synthesized by . ZJB2016050 with high-yield amphotericin B (AmB) was obtained by traditional mutagenesis using ATCC14899 as the original strain. The differences in the characterization of the two strains were found in color, mycelium morphology, and AmB yield. Subsequent comparative transcriptome explained the yield differences between the two strains. Pathways including the carbohydrate metabolic pathway and the secondary product synthesis pathway were targeted. The upregulation of glucokinase, phosphoglycerate mutase, and pyruvate dehydrogenase accelerates the consumption of glucose and has great effects on the accumulation of precursors. One of the competitive secondary metabolites of the polyketone synthetase (PKS) II type sapromomycin analog synthesis gene cluster was downregulated, which competes for malonyl-CoA. Five PKS modules (except for the first module amphA) of the amphotericin synthetic gene cluster in the high-yielding strain were downregulated, which resulted in the total amphotericin A (AmA) and AmB of ZJB2016050 being less than that of the wild-type ATCC14899. Combined with gene differential expression in the pentose phosphate pathway and the reaction mechanism of the ER5 domain, the reason that ZJB2016050 preferred to synthesize AmB was probably related to intracellular reduction.
抗生素在人类健康中发挥着重要作用。大多数抗生素源自微生物次级代谢产物。两性霉素是一种由……合成的多烯大环内酯类抗生素。以ATCC14899为原始菌株，通过传统诱变获得了高产两性霉素B（AmB）的ZJB2016050。发现这两种菌株在颜色、菌丝形态和AmB产量方面存在特征差异。随后的比较转录组分析解释了两种菌株之间的产量差异。靶向了包括碳水化合物代谢途径和次级产物合成途径在内的途径。葡萄糖激酶、磷酸甘油酸变位酶和丙酮酸脱氢酶的上调加速了葡萄糖的消耗，并对前体物质的积累有很大影响。聚酮合酶（PKS）II型腐霉素类似物合成基因簇的一种竞争性次级代谢产物被下调，该产物与丙二酰辅酶A竞争。高产菌株中两性霉素合成基因簇的五个PKS模块（除第一个模块amphA外）被下调，这导致ZJB2016050的总两性霉素A（AmA）和AmB低于野生型ATCC14899。结合磷酸戊糖途径中的基因差异表达和ER5结构域的反应机制，ZJB2016050优先合成AmB的原因可能与细胞内还原有关。
