|
KMID : 1024620190390010013
|
|
Food Science of Animal Resources
2019 Volume.39 No. 1 p.13 ~ p.22
|
|
|
Construction of a Bile-responsive Expression System in Lactobacillus plantarum
|
|
Chae Jong-Pyo
Pajarillo Edward Alain B.
Hwang In-Chan
Kang Dae-Kyung
|
|
|
Abstract
|
|
|
|
This study aimed to develop a bile-responsive expression system for lactobacilli. The promoters of four genes, encoding phosphoenolpyruvate-dependent sugar phosphotransferase (mannose-specific), L-lactate dehydrogenase (LDH), HPr kinase, and D-alanine-D-alanine ligase, respectively, which were highly expressed by bile addition in Lactobacillus johnsonii PF01, were chosen. Each promoter was amplified by polymerase chain reaction and fused upstream of the β-glucuronidase gene as a reporter, respectively. Then, these constructs were cloned into E. coli-Lactobacillus shuttle vector pULP2, which was generated by the fusion of pUC19 with the L. plantarum plasmid pLP27. Finally, the constructed vectors were introduced into L. plantarum for a promoter activity assay. The LDH promoter showed the highest activity and its activity increased 1.8-fold by bile addition. The constructed vector maintained in L. plantarum until 80 generations without selection pressure. A bile-responsive expression vector, pULP3-PLDH, for Lactobacillus spp. can be an effective tool for the bile-inducible expression of bioactive proteins in intestine after intake in the form of fermented dairy foods.
|
|
|
KEYWORD
|
|
|
bile-responsive , expression , Lactobacillus , plasmid
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|