Purification, properties, and N-terminal amino acid sequences of guar gum-degrading enzyme from Bacillus circulans K-1.
A guar gum-degrading enzyme of the newly isolated Bacillus circulans K-1 was purified to an electrophoretically homogeneous state. The molecular weight of the purified enzyme was 62,000 by SDS-PAGE. The purified enzyme was separated into a least six isozymes by isoelectric focusing and the pI of these isozymes were 5.4, 5.5, 5.6, 5.8, 6.0, and 6.2, respectively. The N-terminal amino acid sequences of the typical three of these proteins were all the same, Ala-Ser-Gly-Phe-Tyr-Val-Ser-Gly-Thr-Lys-Leu-Asp-Ala-Thr-Gly-Gln-Pro-Phe- Val- Met-Arg. The enzyme was most active at pH 6.9 and at 64 degrees C. The enzyme was activated slightly by Al3+ and inhibited strongly by Sn2+ and Zn2+, N-bromosuccinimide, 2-mercaptoethanol, and ethylenediamine-tetraacetic acid.
Yosida
S
S
Research Institute of Technology, Konoike Construction Co., Ltd., Osaka, Japan.
Sako
Y
Y
Uchida
A
A
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology*; D005685:Galactans / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007477:Ions; D007526:Isoelectric Point; D008351:Mannans / Q000378:metabolism*; D008361:Mannosidases / Q000737:chemistry; D008670:Metals; D008969:Molecular Sequence Data; D008970:Molecular Weight; D053149:Plant Gums; D013379:Substrate Specificity; D013696:Temperature; D044902:beta-Mannosidase
D005685:Galactans; D007202:Indicators and Reagents; D007477:Ions; D008351:Mannans; D008670:Metals; D053149:Plant Gums; C007894:guar gum; D006026:Glycoside Hydrolases; D008361:Mannosidases; C105044:guar gum-degrading enzyme; D044902:beta-Mannosidase