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Purification and characterization of a novel ?-galactosidase from Bacillus sp MTCC 3088

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Purification and characterization of a novel ?-galactosidase from Bacillus sp MTCC 3088
 
Creator Chakraborti, S
Sani, R K
Banerjee, U C
Sobti, R C
 
Subject QR Microbiology
 
Description An extracellular b-galactosidase which catalyzed the production of galacto-oligosaccharide from lactose was harvested
from the late stationary-phase of Bacillus sp MTCC 3088. The enzyme was purified 36.2-fold by ZnCl2 precipitation,
ion exchange, hydrophobic interaction and gel filtration chromatography with an overall recovery of 12.7%.
The molecular mass of the purified enzyme was estimated to be about 484 kDa by gel filtration on a Sephadex G-
200 packed column and the molecular masses of the subunits were estimated to be 115, 86.5, 72.5, 45.7 and 41.2 kDa
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the native enzyme, determined
by polyacrylamide gel electrofocusing, was 6.2. The optimum pH and temperature were 8 and 60°C, respectively.
The Michaelis–Menten constants determined with respect to o-NO2-phenyl-b-D-galactopyranoside and lactose
were 6.34 and 6.18 mM, respectively. The enzyme activity was strongly inhibited (68%) by galactose, the end product
of lactose hydrolysis reaction. The b-galactosidase was specific for b-D anomeric linkages. Enzyme activity was
significantly inhibited by metal ions (Hg2+, Cu2+ and Ag+) in the 1–2.5 mM range. Mg2+ was a good activator. Catalytic
activity was not affected by the chelating agent EDTA.
 
Publisher Springer Science
 
Date 2000
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/849/1/banerjee2000.pdf
Chakraborti, S and Sani, R K and Banerjee, U C and Sobti, R C (2000) Purification and characterization of a novel ?-galactosidase from Bacillus sp MTCC 3088. Journal of Industrial Microbiology and Biotechnology, 24 (1). pp. 58-63. ISSN 1367-5435
 
Relation http://dx.doi.org/10.1038/sj.jim.2900770
http://crdd.osdd.net/open/849/