ZHANG Wen; WANG Chuan; HUANG Cheng Yu; YU Qian; LIU Heng Chuan; ZHANG Chap Wu; PEI Xiao Fang

doi:10.3967/0895-3988.2012.02.012

Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis

ZHANG Wen, WANG Chuan, HUANG Cheng Yu, YU Qian, LIU Heng Chuan, ZHANG Chap Wu, PEI Xiao Fang

文章导航 > Biomedical and Environmental Sciences > 2012 > 25(2): 203-209

ZHANG Wen, WANG Chuan, HUANG Cheng Yu, YU Qian, LIU Heng Chuan, ZHANG Chap Wu, PEI Xiao Fang. Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis[J]. Biomedical and Environmental Sciences, 2012, 25(2): 203-209. doi: 10.3967/0895-3988.2012.02.012

Citation:

ZHANG Wen, WANG Chuan, HUANG Cheng Yu, YU Qian, LIU Heng Chuan, ZHANG Chap Wu, PEI Xiao Fang. Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis[J]. Biomedical and Environmental Sciences, 2012, 25(2): 203-209. doi: 10.3967/0895-3988.2012.02.012

doi: 10.3967/0895-3988.2012.02.012

基金项目: the National Science Foundation of China(30800910)

Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis

Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University,Chengdu 610041, Sichuan, China;Central Hospital of Sichuan Petroleum Bureau, Chengdu 610213, Sichuan, China
Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, Sichuan, China
Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University,Chengdu 610041, Sichuan, China

Funds: the National Science Foundation of China(30800910)

摘要: Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-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 DH5α 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/pMG36eusp-lacZ in both supernatant and cell samples.β-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 β-galactosidase,indicating that the β-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.
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Abstract: Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-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 DH5α 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/pMG36eusp-lacZ in both supernatant and cell samples.β-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 β-galactosidase,indicating that the β-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.
- Gene constructs /
- Gene expression /
- Secretory expression /
- β-Galactosidase /
- Lactococcus lactis

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Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis

doi: 10.3967/0895-3988.2012.02.012

基金项目: the National Science Foundation of China(30800910)

刊出日期: 2012-04-20

关键词:

Gene constructs /

Gene expression /

Secretory expression /

β-Galactosidase /

Lactococcus lactis

摘要: Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-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 DH5α 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/pMG36eusp-lacZ in both supernatant and cell samples.β-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 β-galactosidase,indicating that the β-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.

English Abstract

Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis

Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University,Chengdu 610041, Sichuan, China;Central Hospital of Sichuan Petroleum Bureau, Chengdu 610213, Sichuan, China

Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, Sichuan, China

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

Funds: the National Science Foundation of China(30800910)

Publish Date: 2012-04-20

Keywords:

Abstract: Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-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 DH5α 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/pMG36eusp-lacZ in both supernatant and cell samples.β-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 β-galactosidase,indicating that the β-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.

Citation:

ZHANG Wen, WANG Chuan, HUANG Cheng Yu, YU Qian, LIU Heng Chuan, ZHANG Chap Wu, PEI Xiao Fang. Construction and Secretory Expression of β-Galactosidase Gene from Lactobacillus Bulgaricus in Lactococcus Lactis[J]. Biomedical and Environmental Sciences, 2012, 25(2): 203-209. doi: 10.3967/0895-3988.2012.02.012