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首页> 外文期刊>Biotechnology Progress >L-glutamate enhances the expression of Thermus maltogenic amylase in Escherichia coli
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L-glutamate enhances the expression of Thermus maltogenic amylase in Escherichia coli

机译：L-谷氨酸增强大肠杆菌中产热麦芽淀粉酶的表达

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摘要

Escherichia coli BL21 (DE3) transformed with a thermostable Thermus maltogenic amylase (ThMA), isolated from a Gram-negative bacterium Thermus strain IM6501, grew well and efficiently produced ThMA in a complex medium but not in a chemically defined medium (DM). By supplementing L-glutamate to DM medium, both the specific growth rate and ThMA expression significantly increased. Alterations in the cellular responses of recombinant E. Coli to L-glutamate were analyzed at the protein level by two-dimensional gel electrophoresis and mass spectrometry. The ppGpp synthase (RelA) was significantly reduced in cells grown with L-glutamate and was consistent with the low level of ppGpp, an indicator of stringent response. On the other hand, protein chain elongation factor (EF-Tu) and manganese-containing superoxide dismutase (MnSOD), which protects cells against oxidative damage, was significantly elevated by L-glutamate supplementation. These results indicate that L-glutamate enhances ThMA expression and increases the E. coli growth rate not only by overcoming the stringent response but also by increasing the synthesis of EF-Tu and MnSOD.
机译：从革兰氏阴性细菌Thermus菌株IM6501中分离出的热稳定的Thermus麦芽糖淀粉酶（ThMA）转化的大肠杆菌BL21（DE3）在复杂的培养基中生长良好，并能有效地产生ThMA，但在化学成分确定的培养基（DM）中却无法产生。通过向DM培养基中补充L-谷氨酸，特异性生长速率和ThMA表达均显着增加。通过二维凝胶电泳和质谱分析蛋白质水平上重组大肠杆菌对L-谷氨酸的细胞反应的变化。 ppGpp合酶（RelA）在L-谷氨酸生长的细胞中显着降低，并且与ppGpp的低水平（严格的反应指标）一致。另一方面，补充谷氨酸可显着提高蛋白链延长因子（EF-Tu）和含锰的超氧化物歧化酶（MnSOD），从而保护细胞免受氧化损伤。这些结果表明，L-谷氨酸不仅通过克服严格的应答而且通过增加EF-Tu和MnSOD的合成来增强ThMA表达并增加大肠杆菌的生长速率。

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