Properties of β-Galactosidase from Lactobacillus zymae GU240, an Isolate from Kimchi, and Its Gene Cloning
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Lactobacillus zymae GU240 was previously isolated from Kimchi, a Korean fermented vegetable, as a strong GABA producer. The strain showed β-galactosidase (β-Gal) activity on MRS agar plates with X-gal. When growth and β-Gal activities of GU240 were measured using MRS (glucose, 2%, w/v) and MRSL (lactose, 2%, w/v) broths, cells were found to grow slowly in MRSL, and the β-Gal activity (36 units at 4 h) was lower than that of cells grown in MRS (94 units at 16 h). The highest OD600 value of the culture in MRS was 1.6 at 24 h at 37℃, whereas that of the culture in MRSL was 0.6 at 16 h. β-Gal activity of the culture in MRS reached the maximum (95.6 u/ml) at 16 h, decreased thereafter, and was not detected at 48 h. β-Gal activity for culture in MRSL reached its highest (36 u/ml) at 4 h and decreased gradually, but some activity (11.05 u/ ml) still remained at 72 h. The structural gene encoding β-Gal in L. zymae GU240 was cloned as a 3.1 kb fragment, and DNA sequencing confirmed the presence of complete lacLM genes. lacLM genes from L. zymae GU240 showed 98−99% homologies in nucleotide sequences with other lacLM genes from L. brevis. Reverse transcription (RT)-PCR confirmed the operon structure of lacLM. The results indicated that L. zymae GU240 might be in the process of losing the ability to grow rapidly on lactose-containing medium, such as milk, due to adaptations to plant environments, including kimchi.
Abstract
Lactobacillus zymae GU240 was previously isolated from Kimchi, a Korean fermented vegetable, as a strong GABA producer. The strain showed β-galactosidase (β-Gal) activity on MRS agar plates with X-gal. When growth and β-Gal activities of GU240 were measured using MRS (glucose, 2%, w/v) and MRSL (lactose, 2%, w/v) broths, cells were found to grow slowly in MRSL, and the β-Gal activity (36 units at 4 h) was lower than that of cells grown in MRS (94 units at 16 h). The highest OD600 value of the culture in MRS was 1.6 at 24 h at 37℃, whereas that of the culture in MRSL was 0.6 at 16 h. β-Gal activity of the culture in MRS reached the maximum (95.6 u/ml) at 16 h, decreased thereafter, and was not detected at 48 h. β-Gal activity for culture in MRSL reached its highest (36 u/ml) at 4 h and decreased gradually, but some activity (11.05 u/ ml) still remained at 72 h. The structural gene encoding β-Gal in L. zymae GU240 was cloned as a 3.1 kb fragment, and DNA sequencing confirmed the presence of complete lacLM genes. lacLM genes from L. zymae GU240 showed 98−99% homologies in nucleotide sequences with other lacLM genes from L. brevis. Reverse transcription (RT)-PCR confirmed the operon structure of lacLM. The results indicated that L. zymae GU240 might be in the process of losing the ability to grow rapidly on lactose-containing medium, such as milk, due to adaptations to plant environments, including kimchi.
참고문헌 (20)
- Juers DH, "LacZ β-galactosidase : structure and function of an enzyme of historical and molecular biological importance", Protein Sci, 21, pp.1792~1807 (2012)
- De Vos WM, "Genetics of lactose utilization in lactic acid bacteria", FEMS Microbiol. Rev, 15, pp.217~237 (1994)
- De Vos WM, "Molecular cloning of lactose genes in dairy lactic streptococci : the phospho-β-galactosidase and β-galactosidase genes and their expression products", Biochimie, 70, pp.461~473 (1988)
- Vaughan EE, "Transcriptional regulation and evolution of lactose genes in the galactose-lactose operon of Lactococcus lactis NCDO2054", J. Bacteriol, 180, pp.4893~4902 (1998)
- Nguyen TH, "Cloning and expression of the β-galactosidase genes from Lactobacillus reuteri in Escherichia coli", J. Biotechnol, 129, pp.581~591 (2007)
- Obst M, "Two genes encoding the β-galactosidase of Lactobacillus sake", Microbiology, 141, pp.3059~3066 (1995)
- Adams RM, "Characterization of two cold-sensitive mutants of the β-galactosidase from Lactobacillus delbruckii subsp. bulgaricus", J. Biol. Chem, 269, pp.5666~5672 (1994)
- Franco D, "Methods on in situ hybridization, immunohistochemistry and beta-galactosidase reporter gene detection", Eur. J. Morphol, 39, pp.3~25 (2001)
- Oliveira C, "Recombinant microbial systems for improved β-galactosidase production and biochemical applications", Biotechnol. Adv, 29, pp.600~609 (2011)
- Platteeuw C, "Food-grade cloning and expression system for Lactococcus lactis", Appl. Environ. Microbiol, 62, pp.1008~1013 (1996)
- Landete JM, "A review of food-grade vectors in lactic acid bacteria : from the laboratory to their application", Crit. Rev. Biotechnol, 37, pp.296~308 (2017)
- Park JY, "Characterization of a glutamate decarboxylase(GAD)gene from Lactobacillus zymae", Biotechnol. Lett, 36, pp.1791~1799 (2014)
- Miller JH, "Experiments in molecular genetics" (1972), Cold Spring Harbor Laboratory Press
- 허경호, "Characterization of a Fibrinolytic Enzyme Secreted by Bacillus amyloliquefaciens CB1 and Its Gene Cloning", Journal of Microbiology and Biotechnology, 23(7), pp.974~983 (2013), 한국미생물·생명공학회[DOI]
- 사현덕, "Characterization of Glutamate Decarboxylase (GAD) from Lactobacillus sakei A156 Isolated from Jeot-gal", Journal of Microbiology and Biotechnology, 25(5), pp.696~703 (2015), 한국미생물·생명공학회[DOI]
- Saier MH, "Catabolite repression and inducer control in Grampositive bacteria", Microbiology, 142, pp.217~223 (1996)
- PARK,RAE-JUN, "Isolation of Lactococcus lactis Strain with β-Galactosidase Activity from Kimchi and Cloning of lacZ Gene from the Isolated Strain", Journal of Microbiology and Biotechnology, 12(1), pp.157~161 (2002), 한국미생물·생명공학회
- Kim TY, "Effect of temperature and a carbon source on the expression of β-galactosidase gene of Lactococcus lactis spp. lactis ATCC7962", J. Microbiol. Biotechnol, 9, pp.201~205 (1999)
- David S, "Leuconostoc lactis β-galactosidase is encoded by two overlapping genes", J. Bacteriol, 174, pp.4475~4481 (1992)
- Chanalia P, "Purification and characterization of β-galactosidase from probiotic Pediococcus acidilactici and its use in milk lactose hydrolysis and galactooligosaccharide synthesis", Bioorg. Chem, 77, pp.176~189 (2018)