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Biomed Environ Sci. 2012 Apr;25(2):203-9.

Construction and secretory expression of beta-galactosidase gene from Lactobacillus bulgaricus in Lactococcus lactis.

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  • 1Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University, Chengdu 610041, Sichuan, China.

Abstract

OBJECTIVE:

This study is to examine the secretion effects of beta-galactosidase in Lactococcus lactis.

METHODS:

The usp45 and beta-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant plasmid pMG36e-usp-lacZ. This recombinant plasmid was transformed into both Escherichia coli DH5alpha and L. lactis MG1363. The enzyme activity, gene sequencing, SDS-PAGE and hereditary stability were assessed and studied.

RESULTS:

The lacZ gene inserted into plasmids pMG36e-usp-lacZ was 99.37% similar to the GenBank sequence, and SDS-PAGE revealed an evident idio-strap at 116 KDa between L. lactis MG1363/pMG36e-usp-lacZ in both supernatant and cell samples. Beta-Galactosidase activity measured 0.225 U/mL in L. lactis pMG36e-usp-lacZ transformants, and its secretion rate was 10%. The plasmid pMG36e-usp-lacZ appeared more stable in MG1363.

CONCLUSION:

The authors concluded that these new recombinant bacteria well expressed and secreted beta-galactosidase, indicating that the beta-galactosidase expression system was successfully constructed, and this might provide a new solution for management of lactose intolerance specifically and promote the use of gene-modified organisms as part of the food-grade plasmid in general.

PMID:
22998828
[PubMed - indexed for MEDLINE]
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