摘　要：谷氨酸转运蛋白（GluA）是一类ATP结合蛋白，可通过调控谷氨酸的主动运输影响细胞内碳代谢、氮代谢和次级代谢，属于ABC转运系统。为研究GluA对放线菌须糖多孢菌（Saccharopolyspora pogona）生长发育及丁烯基多杀菌素生物合成的影响，本研究通过基因编辑技术对编码该蛋白的基因gluA-1分别构建了过表达和敲除菌株。表型分析显示，过表达菌株S. pogona::gluA-1丁烯基多杀菌素产量升高了4.2倍，菌体生长发育对数期延长，菌株最高菌体密度和乙酰辅酶A含量显著提升，但菌丝体形态无明显改变。敲除菌株S. pogona-ΔgluA-1丁烯基多杀菌素产量则下降了37%，菌株生长明显受抑制，乙酰辅酶A含量显著下降，且菌丝体缩短变粗，分支减少。以上结果表明，GluA对须糖多孢菌生长发育和丁烯基多杀菌素生物合成具有一定的促进作用，这为研究其在链霉菌次级代谢过程中的作用奠定了重要基础。
关键词：须糖多孢菌；丁烯基多杀菌素；谷氨酸转运蛋白；生物合成；基因编辑
中文分类号：Q93                                 文献标志码：ADOI：10.3969/j.issn.1007-7146.2024.05.009

Abstract: Glutamate transporter GluA, is a class of ATP-binding proteins that can affect intracellular carbon metabolism, nitrogen metabolism and secondary metabolism by regulating the active transport of glutamate, belonging to the ABC transport system. To study the effects of GluA on the strain growth development and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona, gluA-1 overexpression and deletion strains were constructed by genetic engineering technology. Phenotypic analysis showed that the butenyl-spinosyn production increased by 4.2 times, the bacterial growth logarithmic phase was prolonged, the maximum bacterial density and acetyl-CoA content significantly increased, whereas the mycelium morphology was not significantly changed in overexpressed strain S. pogona::gluA-1. The butenyl-spinosyn production decreased by 37%, the strain growth was inhibited obviously, the acetyl-CoA content significantly decreased, and the mycelium was shortened and thickened, branching was reduced in deletion strain S. pogona-ΔgluA-1. These results indicated that GluA could promote the growth development and the butenyl-spinosyn biosynthesis in S. pogona, which laid an important foundation for the study of GluA's role in the secondary metabolism of Streptomyces.
Key word: Saccharopolyspora pogona; butenyl-spinosyn; glutamate transporter protein; biology synthesis; gene editing
（Acta Laser Biology Sinica, 2024, 33(5): 470-480）