2012

Ksilanaz ve beta (1,3-1,4)- glukanaz enzimlerini kodlayan çift fonksiyonlu genlerin gram-pozitif bakterilerde ekspresyonu

The xylanase and $beta$(1,3-1,4)-Glucanase activities encoding gene from the cellulolytic rumen anaerobe bacterium was cloned into Escherichia/Streptococcus shuttle vector for introduction into Gram-positive bacteria, including the rumen facultative anaerobe bacterium Streptococcus bovis. Activities due to the cloned gene decreased in the stationary phase in batch cultures of S. bovis, reflecting the sensitivity of the cloned enzymes to inactivation in the presence of accumulated lactic acid. Cloned gene activity was detected in the culture supernatant, indicating recognition of the cloned gene signal peptide by Gram-positive bacteria.

Expression of bifunctional genes encoding xylanase and beta (1,3,-1,4)- glucanase in gram-positive bacteria

PDF

___

1. Herbers, K., Flint, H. J., and Sonnewald, V. Apoplastic expressionof the xylanase and β(1,3-1,4) glucanase domains of the xynDgene from Ruminococcus flavefaciensleads to functionalpolypeptides in transgenic tobacco plants. Molecular Breeding.1996; 2: 81-87.
2. Van Rensburg, P., van Zyl, W. H, and Pretorius, I. S. Expressionof the Ruminococcus flavefacienscellodextrian gene inStreptococcus cerevisii. Biotechnol. Letts. 1995; 17: 481-486.
3. Ekinci, M. S., Karaman, M., and Efe, E. Biotechnology, applicationof possible genetic strategies for poultry and ruminant nutrition.International Animal Nutrition Congress “2000” 4-6 September,Isparta/Turkey, 2000; 294-299.
4. Whitehead, T. R, Flint, H. J. Heterologous expression of anendoglucanase gene (endA) from the ruminal anaerobeRuminococcus flavefaciens17 in Streptococcus bovisandStreptococcus sanguis. FEMS Microbiol Lett. 1995; 126: 165-170
5. Jones, B. A., Mucke, R. F., and Ricke S. C. Selection andapplication of Streptococcus bovisas a silage inoculant. Appl.Environ. Microbiol. 1991; 57: 3000-3005.
6. Satoh, E., Ho, Y., Sasaki, Y., and Sasaki, T. Application of theextracellular a-amylase gene from Streptococcus bovis148 toconstruction of a separation water for yogurt starter strains. Appl.Environ. Microbiol. 1997; 63: 4593-4596.
7. Ekinci, M.S., McCrae, S.I., and Flint, H.J. Isolation andoverexpression of a gene encoding an extracellular β-(1,3-1,4)-glucanase from Streptococcus bovisJB1. Appl. Environ. Microbiol.1997; 63: 3762-3756.
8. Hespell, R. B., and Whitehead, T. R. Introduction of Tn916 andpAMβ1 into Streptococcus bovisJB1 by conjungation. Appl.Environ. Microbiol. 1991; 57: 2710-2713.
9. Özcan, N. and Ekinci, M. S. Studies on polysaccharide degradingenzymes encoding genes. Challenges For Microbial DigestiveEcology. 2ndJoint INRA-RRI gastrointestinal tract Microbiologysymposium, Clermont-Ferrand, France, in “Reproduction NutritionDevelopment. 2000; 40: 221”.
10. Mercer, D. K., Melville, C. M., Scott, K. P., and Flint, H. J. Naturalgenetic transformation in the rumen bacterium Streptococcusbovis JB1. FEMS Microbiol. Letts. 1999; 179: 485-490.
11. Macrina, F. L., Tobian, J. A., Jones, K. R, and Evans, R. P, ClewellD. B. A cloning vector able to replicate in Escherichia coliandStreptococcus sanguis. Gene. 1982; 19: 345-353.
12. Simon, D., and Chopin, A. Construction of a vector plasmid familyand its use for molecular cloning in Streptococcus lactis.Biochimie. 1988; 70: 559-566.
13. Hobson, P. N. Rumen bacteria. In “Methods in microbiology” 3Bed Norris JR, Ribbons DW. London, Academic Press, 1969; 133-149.
14. Whitehead, T. R. Analyses of the gene and amino acid sequence ofthe Prevotella (Bacteroides) ruminicola23 xylanase revealsunexpected homology with endoglucanases from other genera ofbacteria. Curr. Microbiol. 1993; 27: 27-34.
15.Holo, H., and Nes, I. F. High frequency transformation, byelectroporation, of Lactococcus lactis subsp. Cremorisgrown withglycine in osmotically stabilized media. Appl. Environ. Microbiol.1989; 55: 3119-3123.
16. Cruz-Rodz, A. L, and Gilmore, M. S. Electroporation of glycine-treated Enterococcus faecalis, In “Genetics and Molecular Biologyof Streptococci, Lactococci andEnterococci” Eds. Dunny, G.M.,Cleary, P.P., McKay, L.L. ASM Washington, D.C. 1991; 300.
17.Lever, M. Carbohydrate determination with 4-hydroxybenzoic acidhydrazide (PAHBAH): effect of bismuth on the reaction. Anal.Biochem. 1977; 81: 21-27.
18. Teather, R. M., and Wood, P. J. Use of Congo Red-polysaccharideinteractions in enumeration and characterisation of cellulolyticbacteria from the bovine rumen. Appl. Environ. Microbiol. 1982;43: 777-780
19. Flint, H. J, Martin, J., McPherson, C. A., Daniel, A. S., and Zhang,J-X. A bifunctional enzyme, with separate xylanase and β(1,3-1,4)-glucanase domains, encoded by the xynDgene ofRuminococcus flavefaciens.J. Bacteriol. 1993; 175: 2943-2951.
20. Kirby, J., Martin, J. C., Daniel, A. S., and Flint, H. J. Dockerin-likesequences in cellulases and xylanases from the rumen cellulolyticbacterium Ruminococcus flavefaciens. FEMS Microbiol. Lett.1997; 149: 213-219.
21. Scott, K. P, Mercer, D. K, Glover, L. A, and Flint H. J. The greenfluorescent protein as a visible marker for lactic acid bacteria incomplex anaerobic ecosystems. FEMS Microbiol. Ecol. 1998; 26:219-230.
22. Deguchi, H., Watanabe Y., Sasaki T., Matsuda T., Shimizu S., andOhmiya K. Purification and properties of the endo-1,4-β-glucanasefrom Rumnococcus albusand its gene product in Escherichia coli.J. Ferment. Bioeng. 1991; 71: 221-225.
23. Erickson, P. R., and Herzberg, M. C. Evidence for the covalentlinkage of carbohydrate polymers to a glycoprotein fromStreptococcus sanguis. J. Biol. Chem. 1993; 268: 23780-23783