OPTIMIZATION OF ELECTROPORATION MEDIATED TRANSFORMATION OF LACTOBACILLUS PLANTARUM FOR INDUSTRIAL EXPLOITATION
Keywords:
Lactobacillus plantarum, Transformation, Electroporation, Lithium acetateAbstract
Objective: The objective of our study was to evaluate different established electrotransformation protocols for their suitability on Lactobacillus plantarum.
Methods: We tested strategies of electrical parameters in addition to already published electroporation methods proven to be successful in other Lactobacillus strains.
Results: Our strain differed highly in transformation efficiencies and optimal growth conditions. Among the various methods performed, we observed high-efficiency protocol yielded 8X106 transformants, and the optimum electroporation parameter was found to be at 1500V.
Conclusion: Independent of electroporation method Lactobacillus plantarum strain remained reluctant to high-efficiency transformation protocol.Downloads
References
Pot B, Tsakalido E. Lactobacillus molecular biology: from genomics to probiotics. Norfolk: Caister Academic Press; 2009.
Hammes WP, Hertel C. The Genera Lactobacillus, and Carnobacterium. The Prokaryotes. New York: Springer; 2003.
Tamime AY. Probiotic dairy products. Oxford: Blackwell Publishing; 2005.
Vemuri PK, Velampaty RHP, Tipparaju SL. Probiotics: a novel approach in improving the values of human life. Int J Pharm Pharm Sci 2014;6:41-3.
De Mey M, Maertens J, Boogmans S, Soetaert WK, Vandamme EJ, Cunin R, et al. Promoter knock-in: a novel rational method for the fine tuning of genes. BMC Biotechnol 2010;10:26.
Hols P, Slos P, Dutot P, Reymund J, Chabot P, Delplace B. Efficient secretion of the model antigen M6-gp41E in Lactobacillus plantarum NCIMB 8826. Microbiology 1997;143:2733-41.
Wells JM, Wilson PW, Norton PM, Gasson MJ, Le Page RW. Lactococcus lactis: highâ€level expression of tetanus toxin fragment C and protection against lethal challenge. Mol Microbiol 1993;8:1155-62.
Xie TD, Sun L, Tsong TY. Study of mechanisms of electric field-induced DNA transfection. I. DNA entry by surface binding and diffusion through membrane pores. Biophysical J 1990;58:13.
Dunny GM, Lee LN, LeBlanc DJ. Improved electroporation and cloning vector system for gram-positive bacteria. Appl Environ Microbiol 1991;57:1194-201.
Bringel F, Hubert JC. Optimized transformation by electroporation of Lactobacillus plantarum strains with plasmid vectors. Appl Microbiol Biotechnol 1990;33:664-70.
Aukrust TW, Brurberg MB, Nes IF. Transformation of Lactobacillus by electroporation. Electroporation Protocols Microorganisms; 1995. p. 201-8.
Holo H, Nes IF. High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl Environ Microbiol 1989;55:3119-23.
Aukrust T, Blom H. Transformation of Lactobacillus strains used in meat and vegetable fermentations. Food Res Int 1992;25:253-61.
Ahrné S, Molin G, Axelsson L. Transformation ofLactobacillus reuteri with electroporation: studies on the erythromycin resistance plasmid pLUL631. Curr Microbiol 1992;24:199-205.
Kumar Vemuri P, Veeravalli S. Expression, purification and characterization of human recombinant galectin 3 in pichia pastoris. Iran J Biotechnol 2014;12:3-8.
Wards BJ, Collins DM. Electroporation at elevated temperatures substantially improves transformation efficiency of slow-growing mycobacteria. FEMS Microbiol Lett 1996;145:101-5.
Leenhouts KJ, Tolner B, Bron S, Kok J, Venema G, Seegers JF. Nucleotide sequence and characterization of the broad-host-range lactococcal plasmid pWVO1. Plasmid 1991;26:55-66.
Spath K, Heinl S, Egger E, Grabherr R. Lactobacillus plantarum and Lactobacillus buchneri as expression systems: evaluation of different origins of replication for the design of suitable shuttle vectors. Mol Biotechnol 2012;52:40-8.
Gietz RD, Woods RA. Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 2002;350:87-96.
Papagianni M, Avramidis N, Filioussis G. High-efficiency electrotransformation of Lactococcus lactis spp. lactis cells pretreated with lithium acetate and dithiothreitol. BMC Biotechnol 2007;7:1.
Palomino MM, Allievi MC, Prado-Acosta M, Sanchez-Rivas C, Ruzal SM. New method for electroporation of Lactobacillus species grown in high salt. J Microb Med 2010;83:164-7.
Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 2002;40:2153-62.