Purification and properties of a high-molecular-weight, alkaline exopolygalacturonase from a strain of Bacillus

被引：36

作者：

Kobayashi, T ^{[1
]}

Higaki, N ^{[1
]}

Suzumatsu, A ^{[1
]}

Sawada, K ^{[1
]}

Hagihara, H ^{[1
]}

Kawai, S ^{[1
]}

Ito, S ^{[1
]}

机构：

[1] Kao Corp, Tochigi Res Labs, Haga, Tochigi 3213497, Japan

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2001年 / 29卷 / 01期

关键词：

exopolygalacturonase; pectic enzyme; Bacillus;

D O I：

10.1016/S0141-0229(01)00355-6

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An exopolygalacturonase [exo-PG; poly (1,4-alpha -D-galacturonide) digalacturonohydrolase, EC 3.2.1.82] was found in a culture of Bacillus sp. strain KSM-P576. The purified exo-PG had a molecular weight of approximately 115,000 and an isoelectric point of pH 4.6. The N-terminal amino acid sequence was Thr-Glu-Val-Ser-Pro-Lys-Ser-Pro-Ala-Ser-Pro-Val. Maximum activity toward polygalacturonic acid (PGA) was observed at 55 degreesC and pH 8.0 in 100 mM Tris-HCl buffer. The exo-PG was quite stable in various pH buffers between pH 6 and 12 when incubated at 30 degreesC for 1 h. Mg2+ Mn2+ Pd2+ and Ca2+ ions stimulated the enzyme activity. The exo-PG released digalacturonic acid from PGA, tri-, tetra-, and penta-galacturonic acids. The apparent K-m values for oligogalacturonic acids were almost identical, and k(cat) values increased with the chain length of the substrates. (C) 2001 Elsevier Science Inc. All rights reserved.

引用

页码：70 / 75

页数：6

共 32 条

[21] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[22]

Maceira FIG, 1997, FEMS MICROBIOL LETT, V154, P37, DOI 10.1016/S0378-1097(97)00298-X

[23] USE OF DINITROSALICYLIC ACID REAGENT FOR DETERMINATION OF REDUCING SUGAR [J].

MILLER, GL .

ANALYTICAL CHEMISTRY, 1959, 31 (03) :426-428

[24] Novel exopolygalacturonases produced by Alternaria mali [J].

Nozaki, K ;

Miyairi, K ;

Hozumi, S ;

Fukui, Y ;

Okuno, T .

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1997, 61 (01) :75-80

[25] A new high-alkaline and high-molecular-weight pectate lyase from a Bacillus isolate:: Enzymatic properties and cloning of the gene for the enzyme [J].

Ogawa, A ;

Sawada, K ;

Saito, K ;

Hakamada, Y ;

Sumitomo, N ;

Hatada, Y ;

Kobayashi, T ;

Ito, S .

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2000, 64 (06) :1133-1141

[26] Pectinase in papermaking: solving retention problems in mechanical pulps bleached with hydrogen peroxide [J].

Reid, I ;

Ricard, M .

ENZYME AND MICROBIAL TECHNOLOGY, 2000, 26 (2-4) :115-123

[27]

Rijssel M. van, 1993, Applied and Environmental Microbiology, V59, P828

[28] PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR PECTINOLYTIC ENZYMES PRODUCED BY SCLEROTINIA-SCLEROTIORUM [J].

RIOU, C ;

FREYSSINET, G ;

FEVRE, M .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1992, 58 (02) :578-583

[29]

ROMBOUTS FM, 1980, ECON MICROBIOL, V5, P227

[30] PECTIN, PECTINASE, AND PROTOPECTINASE - PRODUCTION, PROPERTIES, AND APPLICATIONS [J].

SAKAI, T ;

SAKAMOTO, T ;

HALLAERT, J ;

VANDAMME, EJ .

ADVANCES IN APPLIED MICROBIOLOGY, VOL 39, 1993, 39 :213-294

← 1 2 3 4 →