title: Applications of β-gal-III isozyme from Bacillus coagulans RCS3, in lactose hydrolysis.
abstract: Bacillus coagulans RCS3 isolated from hot water springs secreted five isozymes i.e. β-gal I-V of β-galactosidase. β-gal III isozyme was purified using DEAE cellulose and Sephadex G 100 column chromatography. Its molecular weight characterization showed a single band at 315kD in Native PAGE, while two subunits of 50.1 and 53.7 kD in SDS PAGE. β-Gal III had pH optima in the range of 6-7 and temperature optima at 65°C. It preferred nitro-aryl-β-d-galactoside as substrate having K(m) of 4.16 mM with ONPG. More than 85% and 80% hydrolysis of lactose (1-5%, w/v) was recorded within 48 h of incubation at 55°C and 50°C respectively and pH range of 6-7. About 78-86% hydrolysis of lactose in various brands of standardized milk was recorded at incubation temperature of 50°C. These results marked the applications of β-gal III in processing of milk/whey industry.
authors.affiliation: Dept. of Biotechnology, GGDSD College, Chandigarh, India. batranavneet@gmail.com
authors.forename: Navneet
authors.initials: N
authors.lastname: Batra
authors.forename: Jagtar
authors.initials: J
authors.lastname: Singh
authors.forename: Amit
authors.initials: A
authors.lastname: Joshi
authors.forename: Sonu
authors.initials: S
authors.lastname: Bhatia
mesh_terms: D000818:Animals; D001407:Bacillus / Q000737:chemistry / Q000201:enzymology* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D019649:Food Industry / Q000379:methods*; D045482:Hot Springs / Q000382:microbiology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D007785:Lactose / Q000378:metabolism*; D008892:Milk / Q000378:metabolism*; D008970:Molecular Weight; D021122:Protein Subunits; D013379:Substrate Specificity; D013696:Temperature; D001616:beta-Galactosidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D007527:Isoenzymes; D021122:Protein Subunits; D001616:beta-Galactosidase; D007785:Lactose