Preparation of γ-aminobutyric acid using E-coli cells with high activity of glutamate decarboxylase

被引:69
作者
Plokhov, AY [1 ]
Gusyatiner, MM
Yampolskaya, TA
Kaluzhsky, VE
Sukhareva, BS
Schulga, AA
机构
[1] Mosagrogen Joint Stock Co, Moscow 113545, Russia
[2] Moscow Genet & Select Ind Microorganisms Inst, Moscow 113545, Russia
[3] Russian Acad Sci, Engelhardt Inst Mol Biol, Moscow 117984, Russia
关键词
gamma-aminobutyric acid; glutamate decarboxylase; Escherichia coli; pyridoxal; 5-phosphate;
D O I
10.1385/ABAB:88:1-3:257
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
gamma-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter synthesized in the central nervous system from glutamate by glutamate decarboxylase (GAD). It has applications in the production of many drugs. The technology of GABA synthesis by treating L-glutamic acid with the cells of the gene-engineered GAD superproducer strain of Escherichia coli GADK10 was developed. Cell growing in the presence of 0.02 mM pyridoxal phosphate (PLP) causes the 2- to 2.5-fold increase of total productivity of the cells. The best way to prep are the cells for the reaction was their thermal activation by pretreatment for 1 h at 53 degrees C. The optimal conditions for this reaction were 37 degrees C and pH 4.6. The rate of the enzymatic reaction is the function of acetate concentration with the maximum at 0.5 M acetate. The total amount of GABA synthesized using 1 g of wet cells reached 23-25 g. The final concentration of GABA in the reaction medium was 280-300 g/L. The yield of the product was about 99%.
引用
收藏
页码:257 / 265
页数:9
相关论文
共 15 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   THE PREPARATION OF GAMMA-AMINOBUTYRIC ACID BY ENZYMATIC DECARBOXYLATION OF L-GLUTAMIC ACID [J].
CAMIEN, MN ;
MCCLURE, LE ;
LEPP, A ;
DUNN, MS .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1953, 43 (02) :378-380
[3]  
DeBiase D, 1996, PROTEIN EXPRES PURIF, V8, P430
[4]  
ERDMAN IE, 1986, ANTIBIOTICS MED BIOT, V4, P249
[5]   GLUTAMATE DECARBOXYLASE - INHIBITION BY MONOCARBOXYLIC ACIDS [J].
FONDA, ML .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1972, 153 (02) :763-768
[6]  
GUBAREV EM, 1960, BIOKHIMIYA, V25, P261
[7]  
KHRISTOFOROV RR, 1995, BIOCHEM MOL BIOL INT, V36, P77
[8]   CHEMICAL NATURE OF SYNAPTIC TRANSMISSION IN VERTEBRATES [J].
KRNJEVIC, K .
PHYSIOLOGICAL REVIEWS, 1974, 54 (02) :418-540
[9]   IMMOBILIZATION OF GLUTAMATE-DECARBOXYLASE AND THE PREPARATION OF AN ENZYME COLUMN FOR THE SYNTHESIS OF GAMMA-[N-13]AMINOBUTYRIC ACID [J].
LAMBRECHT, RHD ;
SLEGERS, G ;
MANNENS, G ;
CLAEYS, A .
ENZYME AND MICROBIAL TECHNOLOGY, 1987, 9 (04) :221-224
[10]   GAMMA-AMINOBUTYRIC ACID AND NERVOUS-SYSTEM FUNCTION - PERSPECTIVE [J].
ROBERTS, E .
BIOCHEMICAL PHARMACOLOGY, 1974, 23 (19) :2637-2649