Cell disruption optimization and covalent immobilization of beta-D-galactosidase from Kluyveromyces marxianus YW-1 for lactose hydrolysis in milk.
beta-D-galactosidase (EC 3.2.1.23) from Kluyveromyces marxianus YW-1, an isolate from whey, has been studied in terms of cell disruption to liberate the useful enzyme. The enzyme produced in a bioreactor on a wheat bran medium has been successfully immobilized with a view to developing a commercially usable technology for lactose hydrolysis in the food industry. Three chemical and three physical methods of cell disruption were tested and a method of grinding with river sand was found to give highest enzyme activity (720 U). The enzyme was covalently immobilized on gelatin. Immobilized enzyme had optimum pH and temperature of 7.0 and 40 degrees C, respectively and was found to give 49% hydrolysis of lactose in milk after 4 h of incubation. The immobilized enzyme was used for eight hydrolysis batches without appreciable loss in activity. The retention of high catalytic activity compared with the losses experienced with several previously reported immobilized versions of the enzyme is significant. The method of immobilization is simple, effective, and can be used for the immobilization of other enzymes.
Puri
Munish
M
Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, India. mpuri@pbi.ac.in
Gupta
Shivani
S
Pahuja
Parveen
P
Kaur
Aneet
A
Kanwar
J R
JR
Kennedy
J F
JF
D000818:Animals; D004800:Enzymes, Immobilized / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D005780:Gelatin / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007716:Kluyveromyces / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D007785:Lactose / Q000378:metabolism*; D008892:Milk / Q000378:metabolism*; D010539:Permeability / Q000187:drug effects; D013696:Temperature; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*
D004800:Enzymes, Immobilized; D005780:Gelatin; D001616:beta-Galactosidase; D007785:Lactose