Purification and stabilization of a monomeric isocitrate dehydrogenase from Corynebacterium glutamicum

被引：20

作者：

Bai, C ^{[1
]}

Fernandez, E ^{[1
]}

Yang, H ^{[1
]}

Chen, RD ^{[1
]}

机构：

[1] Univ Saskatchewan, Coll Med, Dept Biochem, Saskatoon, SK S7N 5E5, Canada

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1999年 / 15卷 / 03期

基金：

英国医学研究理事会;

关键词：

D O I：

10.1006/prep.1999.1034

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Monomeric isocitrate dehydrogenase was expressed in Corynebacterium glutamicum cells harboring pEK-icdES1, a plasmid carrying the gene for the enzyme. Two- to three-fold higher expression levels of the recombinant enzyme were observed in such cells when grown in fermenters, compared to those grown in shaker incubators. The enzyme was purified to homogeneity by ammonium sulfate fractionation, Sephadex G-150 gel filtration, FPLC Mono Q anion-exchange chromatography, and affinity gel chromatography. Approximately 4 mg of 98% pure recombinant enzyme was obtained per liter of bacterial culture. Our results also include optimum buffer conditions for purification and storage of the enzyme. (C) 1999 Academic Press.

引用

页码：344 / 348

页数：5

共 10 条

[1] MUTAGENESIS AND LAUE STRUCTURES OF ENZYME INTERMEDIATES - ISOCITRATE DEHYDROGENASE [J].

BOLDUC, JM ;

DYER, DH ;

SCOTT, WG ;

SINGER, P ;

SWEET, RM ;

KOSHLAND, DE ;

STODDARD, BL .

SCIENCE, 1995, 268 (5215) :1312-1318

[2] PURIFICATION AND COMPARATIVE PROPERTIES OF THE CYTOSOLIC ISOCITRATE DEHYDROGENASES (NADP) FROM PEA (PISUM-SATIVUM) ROOTS AND GREEN LEAVES [J].

CHEN, R ;

LEMARECHAL, P ;

VIDAL, J ;

JACQUOT, JP ;

GADAL, P .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 175 (03) :565-572

[3]

CHEN RD, 1990, PLANT PHYSIOL BIOCH, V28, P411

[4] A HIGHLY-ACTIVE DECARBOXYLATING DEHYDROGENASE WITH RATIONALLY INVERTED COENZYME SPECIFICITY [J].

CHEN, RD ;

GREER, A ;

DEAN, AM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (25) :11666-11670

[5] KINETIC-ANALYSIS OF NAD+-ISOCITRATE DEHYDROGENASE WITH ALTERED ISOCITRATE BINDING-SITES - CONTRIBUTION OF IDH1 AND IDH2 SUBUNITS TO REGULATION AND CATALYSIS [J].

CUPP, JR ;

MCALISTERHENN, L .

BIOCHEMISTRY, 1993, 32 (36) :9323-9328

[6] CLONING, SEQUENCE-ANALYSIS, EXPRESSION, AND INACTIVATION OF THE CORYNEBACTERIUM-GLUTAMICUM-ICD GENE ENCODING ISOCITRATE DEHYDROGENASE AND BIOCHEMICAL-CHARACTERIZATION OF THE ENZYME [J].

EIKMANNS, BJ ;

RITTMANN, D ;

SAHM, H .

JOURNAL OF BACTERIOLOGY, 1995, 177 (03) :774-782

[7] PURIFICATION AND CHARACTERIZATION OF MONOMERIC ISOCITRATE DEHYDROGENASE WITH NADP+-SPECIFICITY FROM VIBRIO-PARAHAEMOLYTICUS Y-4 [J].

FUKUNAGA, N ;

IMAGAWA, S ;

SAHARA, T ;

ISHII, A ;

SUZUKI, M .

JOURNAL OF BIOCHEMISTRY, 1992, 112 (06) :849-855

[8] STRUCTURE OF A BACTERIAL ENZYME REGULATED BY PHOSPHORYLATION, ISOCITRATE DEHYDROGENASE [J].

HURLEY, JH ;

THORSNESS, PE ;

RAMALINGAM, V ;

HELMERS, NH ;

KOSHLAND, DE ;

STROUD, RM .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (22) :8635-8639

[9] GENES ENCODING 2 ISOCITRATE DEHYDROGENASE ISOZYMES OF A PSYCHROPHILIC BACTERIUM, VIBRIO SP STRAIN ABE-1 [J].

ISHII, A ;

SUZUKI, M ;

SAHARA, T ;

TAKADA, Y ;

SASAKI, S ;

FUKUNAGA, N .

JOURNAL OF BACTERIOLOGY, 1993, 175 (21) :6873-6880

[10] PURIFICATION AND CHARACTERIZATION OF A MONOMERIC ISOCITRATE DEHYDROGENASE WITH DUAL COENZYME SPECIFICITY FROM THE PHOTOSYNTHETIC BACTERIUM RHODOMICROBIUM-VANNIELII [J].

LEYLAND, ML ;

KELLY, DJ .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1991, 202 (01) :85-93

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