Purification and some properties of glucose isomerase from Bacillus megaterium
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Abstract
The objective of this study is to produce and purify glucose isomerase (GI) from Bacillus megaterium and to determine some of its properties. Soil sample was collected from cassava starch processing site and used immediately for bacterial isolation. Selected isolate produced the best GI activity in a preliminary test. The isolate was grown in media containing various carbon, nitrogen and metal salts for enzyme production. Among the carbon sources tested, Galactose gave the best yield (1.1 U/mg protein). Peptone was the best nitrogen source and caused the production of 0.85 U/mg protein. Of the tested metal salts, MgSO4.7H2O caused the production of the best enzyme activity of 0.81 U/mg protein. The GI was purified by precipitation with (NH4)2SO4 and chromatography on diethyl amino-ethyl (DEAE) – cellulose and DEAE-sephadex G-200 column. Thereafter, the enzyme activity was determined with the optimum temperature and pH at 40oC and 6.0 respectively. The enzyme was also characterized with a molecular weight of 65kDa as determined by the SDS – PAGE analysis. The enzyme could be applied in the production of high fructose corn syrup.
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References
Aiyer, D.V.D. (2004). Some properties of glucose isomerase from Streptomyces flavogriseus. African J. Biotechnol., 3(10): 519 – 522.
Angardi, V. and Calik, P. (2013). Beet molasses based exponential feeding strategy for thermostable glucose isomerase production by recombinant Escherichia coli BL21 (DE3). J. Chem. Technol. Biotechnol. 88(5): 845-852.
Bhosale S.H., Rao M.B., and Deshpande V.V. (1996). Molecular and industrial aspects of glucose isomerase. Microbiol. Rev., 60: 280 – 300.
Chen, W.P., Anderson A.W. and Han Y.W. (1979). Purification of glucose isomerase by Streptomyces flarogriseus. Appl. Environ. Microbiol. 37: 324-331.
Danno, G.L. (1970). Studies on D-glucose-isomerizing enzyme from Bacillus coagulans strain HN-68. Agric. Biol. Chem. 34: 1805-1814.
Deshmukh, S.S., Deshpande, M.V. and Shankar, V. (1994). Medium optimization for the production of glucose isomerase from thermophilic Streptommsces thermonitrificans. World J. Microbiol Biotechnol. 10: 264-267.
Dhungel, B., Subedi, M., Tiwari, K.B., Shrestha, U.T., Pokhrel, S. and Agrawal, V.P. (2007). The thermostable glucose isomerase from psychrotolerant Streptomyces species. Int. J. Life Sci 1: 6-10.
Hartley, B.S., Hanlon, N., Jackson, R.J. and Rangarajan, M. (2000). Glucose isomerase: Insights into protein engineering for increased themo stability. Biochem. Biophys Acta 154: 294 – 335.
Heo, G.Y., Kim, W.C., Joo, G.J., Kwak, Y.Y. (2008). Deletion of xylR gene enhances expression of xylose isomerase in Streptomyces lividans TK24 J. Microbiol. Biotechnol., 18(5): 837 – 44.
Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley J.T. and Williams, S.T. (1994). Bergey’s Manual of Determinative Bacteriology. 9th Edn. Baltimore: Williams and Wilkins.
Kasumi T., Hayashi, K. and Tsumura, N. (1980). Purification and enzymatic properties of glucose isomerase from Streptomyces griseofuscus S-41. Agric. Biol. Chem. 45: 1087-1095.
Kitada, M., Dobshi, Y. and Horikoshi, K. (1989). Enzymatic properties of purified D-xylose isomerase from a thermophilic alkalophile Bacillus TX-3. Agric Biol. Chem. 53(6): 1461-1468.
Kulka, R.G. (1956). Colorimetric estimation of ketopentoses and ketohexoses. Biochem J. 63: 542-548.
Kwakman, J.H.J.M. and Postma, P.W. (1994). Glucose kinase has a regulatory role in catabolite repression in Streptomyces coelicor. J. Bacteriol. 176: 2694-2698.
Laemmli, U.K. (1970). Clearage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227: 680 – 685.
Lawal, A.K., Banjoko, A.M., Banjoko, A.M., Osikoyia, A.F., Olatope, S.O., Kayode, O.F., Etoamihe, M., Emoleila., I., Alebiosu, F.A., Majolagbe, Y.L, Shittu, K.A, Buhari, F. and Dike, E.N. (2012). Production and optimal performance studies of glucose isomerase from agriculture raw material
Global Adv. Research J. Microbiology 1(7): 108-119.
Lee, C. and Zeikus, J.G. (1991). Purification and characterization of thermostable glucose isomerase from Clostridium thermosulfurogenes and
Thermoanaerobacter strain B6A. Biochem. J. 274: 565-571.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R.J. (1951). Protein measurement with folin- phenol reagent. J. Biochem. 193: 265-275.
Pandicurai, N.S.R., Kalaichevan, P.T., Mukesh Kumar D.F. and Gnanaraj M. (2011). Production, optimization and characterization of α-amylase and glucose isomerase producing Bacillus megaterium BPTK5 from cassava waste. International Journal of Current Research, 3(7): 21 – 25.
Ryu, D.Y., Chung, S.H. and Katoh, K. (1977). Performance of the continuous glucose isomerase reactor systems for the production of fructose syrup. Biotechnol. Bioeng. 19: 159-184.
Silva, E.A.B, Souza, A. A.U., Rodrigues A.E. and Guelli, S.M.A. (2006). Glucose isomerisation in simulated moving bed reactor by glucose isomerase. Braz. Arch. Biol. Technol. 49:491-502.
Sriprapundh, D., Vieille, C., and Zeikus, J.G. (2003). Directed evolution of Thermotoga neapolitana xylose isomerase: high activity on glucose at low temperature and low pH. Protein Eng., 16: 683 – 690.
Whitlow, M., Howard, A.J., Finzel, B.C., Poulos, T.L., Winborne, E. and Gilliland, G.L. (1991). A metal-mediated hydride shift mechanism for xylose isomerase based on the 1.6A Streptomyces rubiginosus structures with xylitol and D-xylose. Proteins, 9: 153-173.
Wong H.C., Ting, Y., Lin, H.C., Reichert, F. and Myambo, K. (1991). Genetic organization and regulation of the xylose degradation genes in Streptomyces rubiginosus. J. Bacteriol. 173: 6849-6858.
Yassien, M.A.M. and Jiman-Fatani, A.A.M. (2012). Optimization of glucose isomerase production by Streptomyces albaduncus. Afr J. Microbiol. Res. 6: 2976-2984.