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Purification and characterization of a thermostable α-amylase fromBacillus stearothermophilus

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Abstract

A soil isolate ofBacillus stearothermophilus was found to synthesize thermostable α-amylase. The enzyme was purified to homogeneity by ammonium sulfate fractionation and IECC on DEAE-cellulose column. The purified enzyme was considered to be a monomeric protein with a molar mass of 64 kDa, as determined by SDS-PAGE. The enzyme showed a wide range of pH tolerance and maximum activity at pH 7.0. The temperature tolerance was up to 100 °C with more than 90% catalytic activity: the maximum activity was observed at 50 °C. Divalent metal ions exhibited inhibitory effect on the enzyme activity. However, proteinase inhibitor did not react positively.

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References

  • Bajpai P., Verma N., Bajpai P.K.: Culture conditions for production of thermostable α-amylase byBacillus sp. 25A and properties of the enzyme.J. Microb. Biotechnol.5, 39–52 (1990).

    CAS  Google Scholar 

  • Bandyopadhyay A., Sen S.K., Pal S.C.: Extracellular amylase synthesis byBacillus globisporus Bh-1b.Acta Biotechnol.13, 97–104 (1993).

    Google Scholar 

  • Bose K., Das D.: Thermostable α-amylase production usingBacillus licheniformis NRRL B14368.Ind. J. Exp. Biol.34, 1279–1282 (1996).

    CAS  Google Scholar 

  • Campbell L.L., Cleveland P.D.: Thermostable α-amylase ofBacillus stearothermophilus. IV. Aminoterminal and carboxylterminal amino acid analysis.J. Biol. Chem.236, 2966–2969 (1961).

    PubMed  CAS  Google Scholar 

  • Chakraborty K., Bhattacharyya B.K., Pal S.C., Sen S.K.: Isolation and characterisation of an α-amylase producing strain of thermophilicBacillus.Ind. Biologist30, 30–34 (1998).

    Google Scholar 

  • Chakraborty K., Bhattacharyya B.K., Sen S.K.: Optimisation of fermentation conditions for α-amylase production byBacillus stearothermophilus H2l.Adv. Food Sci.9, 164–167 (1997).

    Google Scholar 

  • David H.M., Gunther H., Roper H.: Catalytic properties ofBacillus megaterium amylase.Starch39, 436–440 (1987).

    Article  CAS  Google Scholar 

  • Galabova D., Velcheva P.: Isolation and purification of α-amylase fromBacillus licheniformis MB 80.Acta Microbiol. Bulg.19, 56–62 (1986).

    CAS  Google Scholar 

  • Hayashi T., Akiba T., Horikoshi K.: Production and purification of new maltohexaose-forming amylase from alkalophilicBacillus sp. H-167.Agric. Biol. Chem.52, 443–448 (1988).

    CAS  Google Scholar 

  • Henderson W.E., Teague W.M.: A kinetic model ofBacillus stearothermophilus α-amylase under process conditions.Starch40, 412–418 (1988).

    Article  CAS  Google Scholar 

  • Jensen B., Olsen J., Allerman K.: Purification of extracellular amylolytic enzymes from the thermophilic fungusThermomyces lanuginosus.Can. J. Microbiol.34, 218–223 (1988).

    Article  CAS  Google Scholar 

  • Kanno M.:A Bacillus acidocaldarius α-amylase that is highly stable to heat under acidic conditions.Agric. Biol. Chem.50, 23–31 (1986).

    CAS  Google Scholar 

  • Koch R., Spreinat T., Lemke K., Antranikian G.: Purification and properties of a hyperthermoactive α-amylase from the archaeobacteriumPyrococcus woesei.Arch. Microbiol.155, 572–578 (1991).

    Article  CAS  Google Scholar 

  • Laemmli U.K.: Cleavage of structure proteins during assembly of the head of bacteriophage T4.Nature69, 646–651 (1970).

    Google Scholar 

  • Makula R., Finnerty W.R.: Microbial assimilation of hydrocarbons. 1. Fatty acids derived fromn-alkanes.J. Bacteriol.95, 2108–2111 (1968).

    PubMed  CAS  Google Scholar 

  • Melasniemi H.: Effect of carbon source on production of thermostable α-amylase, pullulanase and α-glucosidase byClostridium thermohydrosulphuricum.J. Gen. Microbiol.133, 883–890 (1987).

    CAS  Google Scholar 

  • Ramesh M.V., Lonsane B.K.: A novel bacterial thermostable α-amylase system produced under solid state fermentation.Biotechnol. Lett.9, 501–504 (1987).

    Article  CAS  Google Scholar 

  • Sen S.K.: A thermophilicBacillus isolated from the hot spring of Bakreswar.Proc. Plant Sci. Res. Suppl., 204–206 (1987).

  • Soni S.K., Bath K.S., Soni R.: Production of amylases bySaccharomycopsis capsularis in solid state fermentation.Ind. J. Microbiol.36, 157–159 (1996).

    Google Scholar 

  • Srivastava R.A.K., Mathur S.N., Baruah J.N.: Partial purification and properties of thermostable intracellular amylases from a thermophilicBacillus sp. AK-2.Acta Microbiol. Pol.33, 57–66 (1984).

    PubMed  CAS  Google Scholar 

  • Steadman L.B., Thompson C.K., Middaugh R.C., Matsuno K., Vrona S., Lawson Q.E., Lewis V.R.: The effects of surface adsorption on the thermal stability of proteins.Biotechnol. Bioeng.40, 8–15 (1992).

    Article  CAS  PubMed  Google Scholar 

  • Stefanova M.E., Tonkova A.I., Dobreva E.P., Spasova D.I.: Agar gel immobilization ofBacillus brevis cells for production of thermostable α-amylase.Folia Microbiol.43, 42–46 (1998).

    Article  CAS  Google Scholar 

  • Stukalin G.S., Kozlovsky Y.E., Vasylyauskas Y.F., Prozorov A.A.: Thermostable sugarising α-amylase from strainBacillus subtilis 1621: purification and some properties.Biotekhnologiya4, 310–314 (1988).

    CAS  Google Scholar 

  • Tsvetkov V.T., Emanuilova E.I.: Purification and properties of a heat stable α-amylase fromBacillus brevis.Appl. Microbiol. Biotechnol.31, 246–248 (1989).

    Article  CAS  Google Scholar 

  • Wambutt R., Riesenberg D., Kruger M., Schultze M.: Formation of extracellular α-amylase byBacillus subtilis.Z. Allg. Mikrobiol.24, 575–579 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Yankov D., Dobreva E., Beschkov V., Emanuilova E.: Study of optimum conditions and kinetics of starch hydrolysis by means of thermostable α-amylase.Enzyme Microb. Technol.8, 665–666 (1986).

    Article  CAS  Google Scholar 

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Chakraborty, K., Bhattacharyya, B.K. & Sen, S.K. Purification and characterization of a thermostable α-amylase fromBacillus stearothermophilus . Folia Microbiol 45, 207–210 (2000). https://doi.org/10.1007/BF02908945

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