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

被引：17
|
作者：
Uy, D [1 ]
Delaunay, S [1 ]
Engasser, JM [1 ]
Goergen, JL [1 ]
机构：
[1] Inst Natl Polytech Lorraine, CNRS, Lab Sci Genie Chim, F-54505 Vandoeuvre Nancy, France
来源：
JOURNAL OF MICROBIOLOGICAL METHODS | 1999年 / 39卷 / 01期
关键词：
enzymatic assay; pyruvate carboxylase; Corynebacterium glutamicum; glutamate fermentation; biotin; temperature increase;
D O I：
10.1016/S0167-7012(99)00104-9
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A discontinuous lactate dehydrogenase coupled assay is described for the evaluation of the pyruvate carboxylase activity (Pc, EC 6.3.1.1) 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. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码：91 / 96
页数：6
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