一种用于测定谷氨酸棒杆菌谷氨酸发酵过程中丙酮酸羧化酶活性的方法。

A method for the determination of pyruvate carboxylase activity during the glutamic acid fermentation with Corynebacterium glutamicum.

作者信息

Uy D, Delaunay S, Engasser J, Goergen J

机构信息

Laboratoire des Sciences du Génie Chimique-CNRS, Institut National Polytechnique de Lorraine, Vandoeuvre-lès-Nancy, France.

出版信息

J Microbiol Methods. 1999 Dec;39(1):91-6. doi: 10.1016/s0167-7012(99)00104-9.

DOI:10.1016/s0167-7012(99)00104-9

PMID:10579510

Abstract

A discontinuous lactate dehydrogenase coupled assay is described for the evaluation of the pyruvate carboxylase activity (Pc, EC 6.4.1.1) in a glutamate overproducing strain of Corynebacterium glutamicum. After an initial permeabilisation period of the cells, the method consisted of the fluorometric determination of the remaining pyruvate level after transformation into oxaloacetate by the endogenous Pc. The assay was demonstrated to be powerful and enabled the determination of the C. glutamicum Pc activity grown on different carbon sources. Besides, this method was used to assay Pc activity in C. glutamicum 2262 during a temperature triggered glutamate producing process with biotin excess or limitation.

摘要

描述了一种不连续的乳酸脱氢酶偶联测定法，用于评估谷氨酸棒杆菌谷氨酸高产菌株中的丙酮酸羧化酶活性（Pc，EC 6.4.1.1）。在细胞最初的通透化处理期之后，该方法包括通过内源性Pc将丙酮酸转化为草酰乙酸后，对剩余丙酮酸水平进行荧光测定。该测定法被证明是有效的，能够测定在不同碳源上生长的谷氨酸棒杆菌的Pc活性。此外，该方法用于在温度触发的、生物素过量或受限的谷氨酸生产过程中测定谷氨酸棒杆菌2262中的Pc活性。