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Title: -Galactosidase from Lactobacillus acidophilus isolated from fermented ragi (Eleusine coracana)
Authors: Akolkar, S K
Sajgure, A D
Lele, S S
Keywords: Lactobacillus acidophilus
Eleusine coracana
-galactosidase
Issue Date: Apr-2006
Publisher: CSIR
IPC CodeInt. Cl.8 C12N 19/38; C12R1:23
Abstract: Lactobacillus acidophilus was isolated from fermented millet, Eleusine coracana. It was characterized using several biochemical tests. This strain was found to be homofermentative, slime forming and -galactosidase producer. Recovery and characterization of -galactosidase were studied at laboratory scale. Since the enzyme was intracellular various cell lysis methods were studied to achieve maximum enzyme release from cell fragments. Homogenization at a pressure of 2000 psig two passes showed an enzyme release of 2050 U/mL and was found the best method for cell lysis. The cell extract was purified using ultrafiltration and gel permeation chromatography. Specific activity and fold purification of -galactosidase was found to be 568.61 and 21.2, respectively. Kinetic parameters were determined using ONPG (o-nitrophenyl galactopyranoside) as a substrate. Michaeli’s Menten equation was found to fit the reaction of ONPG using -galactosidase from L. acidophilus. Vmax and Km were calculated from the Lineweaver Burk plot. Vmax was found to be 4.94 min-1 and Km to be 0.11 mM. Using gel permeation chromatography, the molecular weight was found to be in the range of 450-500 kDa.
Page(s): 184-188
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.05(2) [April 2006]

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