OBTAINING THE RECOMBINANT α-L-FUCOSIDASE (alfB)
https://doi.org/10.20914/2310-1202-2013-3-178-182
Abstract
In this study the gene α-L-fucosidase (AlfB), amplified by PCR was cloned in the vector plasmid pet23b + sites for EcoR1 - Xho1. Gene expression occurred in the cells of Escherichia coli top 10 under the control of the T7 promoter. The strain-producer recombinant protein α-L-fucosidase was obtained, optimum values of pH and temperature was determined by optimizing the conditions of the enzyme action.
About the Authors
S. V. Kiryanova
Воронежский государственный университет инженерных технологий
Russian Federation
Russian Federation
D. A. Cherenkov
Воронежский государственный университет инженерных технологий
Russian Federation
Russian Federation
A. A. Tolkacheva
Воронежский государственный университет инженерных технологий
Russian Federation
Russian Federation
O. S. Korneeva
Воронежский государственный университет инженерных технологий
Russian Federation
Russian Federation
References
1. Cherenkov, D.A. Fucose : biological role, ways of obtaining and application prospects [Text] / D.A. Cherenkov, Y.A. Rybakov, T.V. Sanina, et al // Biotechnology. - 2010. - № 6. - P. 63-71 .
2. Aminoff, D. Purification and general properties of 1,2 -L-ficosidase from Clostridium Perfringens [Text] / D. Aminoff, K. Furukawa // J. Biol. Chem. – 1970. – V. 245. – P. 1659-1669.
3. Kurimura, Y. Efficient production and purification of extracellular 1,2-alpha-L-fucosidase of Bacillus sp. K40T [Text] / Y. Kurimura // Biotechnol Biochem. - 1995. – V. 59. - № 4. – P. 589-594.
4. Katayama, T. Molecular cloning and characterization of Bifidobacterium bifidum 1,2-α-l-fucosidase (AfcA), a novel inverting glycosidase (Glycoside Hydrolase Family 95) [Text] / Katayama T. // J Bacteriol. – 2004. – V. 186. № 15. – Р. 4885–4893.
5. Sambrook, J. Molecular cloning [Text]: laboratory manual / J. Sambrook, E.F. Fritsch, T. Maniatis. - New York: Cold Spring Harbor Laboratory Press, 1989. – 452 p.
6. Rodriguez-Diaz, J. Utilization of natural fucosylated oligosaccharides by three novel alphaL-fucosidases from a probiotic Lactobacillus casei strain [Text] / J. Rodriguez-Diaz, V. Monedero, M.J. Yebra // Appl. Environ. Microbiol. – 2011. – V. 77. – P. 703-705.
7. Sano, M. Purification and characterization of alpha-L-fucosidase from Streptomyces species [Text] / M. Sano, K. Hayakawa, I. Kato // J. Biol Chem. – 1992. – V. 267. - № 3. – P. 1522-1527.
8. Polygalina, G.V. Determination of enzyme activity [Text]: a guide / G.V. Polygalina, V.S. Cherednychenko, L.V. Rimareva. – M.: DeLee print, 2003. - 375 p.
Review
For citations:
Kiryanova S.V., Cherenkov D.A., Tolkacheva A.A., Korneeva O.S. OBTAINING THE RECOMBINANT α-L-FUCOSIDASE (alfB). Proceedings of the Voronezh State University of Engineering Technologies. 2013;(3):178-182. (In Russ.) https://doi.org/10.20914/2310-1202-2013-3-178-182
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