Data_Sheet_1_Comparative Transcriptome Analysis of Streptomyces nodosus Mutant With a High-Yield Amphotericin B.docx

Antibiotics play an important role in human health. Most antibiotics are derived from microbial secondary metabolites. Amphotericin is a polyene macrolide antibiotic synthesized by Streptomyces nodosus. S. nodosus ZJB2016050 with high-yield amphotericin B (AmB) was obtained by traditional mutagenesis using S. nodosus 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 S. nodosus ZJB2016050 being less than that of the wild-type S. nodosus ATCC14899. Combined with gene differential expression in the pentose phosphate pathway and the reaction mechanism of the ER5 domain, the reason that S. nodosus ZJB2016050 preferred to synthesize AmB was probably related to intracellular reduction.抗生素在人类健康中发挥着至关重要的作用。绝大多数抗生素源自微生物次级代谢产物。两性霉素是由结节链霉菌（Streptomyces nodosus）合成的多烯大环内酯类抗生素。本研究以结节链霉菌ATCC14899为原始菌株，通过传统诱变育种技术获得了高产两性霉素B（AmB）的结节链霉菌ZJB2016050菌株。两株菌株的表型差异体现在菌落颜色、菌丝形态以及AmB产量三个方面。后续的比较转录组分析阐明了两株菌株的产量差异成因，研究靶向了碳水化合物代谢通路与次级产物合成通路。葡萄糖激酶、磷酸甘油酸变位酶与丙酮酸脱氢酶的上调表达加速了葡萄糖的消耗，对前体物质的积累具有显著影响。聚酮合酶（PKS）II型沙霉素类似物合成基因簇所介导的一条竞争性次级代谢通路中的相关基因被下调，该通路会竞争利用丙二酰辅酶A。高产菌株中两性霉素合成基因簇的5个PKS模块（除首个模块amphA外）均被下调，这使得结节链霉菌ZJB2016050的两性霉素A（AmA）与AmB总产量低于野生型菌株结节链霉菌ATCC14899。结合磷酸戊糖通路中的基因差异表达情况与ER5结构域的反应机制，结节链霉菌ZJB2016050更倾向于合成AmB的原因可能与细胞内还原环境相关。