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Metabolic engineering of Lactococcus lactis influence of the overproduction of lipase enzyme

Published online by Cambridge University Press:  24 September 2013

Mohammad Raftari ,
Sobhan Ghafourian  and
Fatimah Abu Bakar
Mohammad Raftari
Affiliation:
Faculty of Food Science and Technology, Universiti Putra Malaysia, UPM 43400, Serdang, Selangor, Malaysia
Sobhan Ghafourian
Affiliation:
Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran
Fatimah Abu Bakar*
Affiliation:
Faculty of Food Science and Technology, Universiti Putra Malaysia, UPM 43400, Serdang, Selangor, Malaysia
*
*For correspondence; e-mail: [email protected]
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Abstract

The dairy industry uses lipase extensively for hydrolysis of milk fat. Lipase is used in the modification of the fatty acid chain length, to enhance the flavours of various chesses. Therefore finding the unlimited source of lipase is a concern of dairy industry. Due to the importance of lipase, this study was an attempt to express the lipase from Burkholderia cepacia in Lactococcus lactis. To achieve this, a gene associated with lipase transport was amplified and subcloned in inducible pNZ8148 vector, and subsequently transformed into Lc. lactis NZ9000. The enzyme assay as well as SDS-PAGE and western blotting were carried out to analysis the recombinant lipase expression. Nucleotide sequencing of the DNA insert from the clone revealed that the lipase activity corresponded to an open reading frame consisting of 1092 bp coding for a 37·5-kDa size protein. Blue colour colonies on nile blue sulphate agar and sharp band on 37·5-kD size on SDS-PAGE and western blotting results confirm the successful expression of lipase by Lc. lactis. The protein assay also showed high expression, approximately 152·2 μg/ml.h, of lipase by recombinant Lc. lactis. The results indicate that Lc. lactis has high potential to overproduce the recombinant lipase which can be used commercially for industrially purposes.

Keywords

Lactococcus lactislipaseoverproduction
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 4 , November 2013 , pp. 490 - 495
Copyright
Copyright © Proprietors of Journal of Dairy Research 2013 

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