Construction and secretory expression of beta-galactosidase gene from Lactobacillus bulgaricus in Lactococcus lactis / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 203-209, 2012.
Article
in English
| WPRIM
| ID: wpr-235534
ABSTRACT
<p><b>OBJECTIVE</b>This study is to examine the secretion effects of beta-galactosidase in Lactococcus lactis.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Base Sequence
/
Beta-Galactosidase
/
DNA Primers
/
Electrophoresis, Polyacrylamide Gel
/
Genetics
/
Lactobacillus
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2012
Type:
Article
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