Thermophilic and Alkalophilic Amylase from Strain Bacillus Marinus MG 12

Authors

  • Dr. Asha B M  Asstistant Professor, Department of Biotechnology, New Horizon College of Engineering, Outer Ring Road, Marathalli, Bengaluru, Karnataka, India
  • Dr. N Sakthivel  Professor, Department of Biotechnology, Pondicherry University
  • Anuradha Singh  Department of Biotechnology, Pondicherry University
  • Dr. H Ananda Vardhan  Associate Professor & HOD, Department of Biotechnology, New Horizon College of Engineering, Outer Ring Road, Marathalli, Bengaluru, Karnataka, India

Keywords:

Amylase, Bacillus, SDS-PAGE, TLC, Kinetics, Specific activity

Abstract

A new bacterial strain MG12 with high amylase activity was characterized as Bacillus marinus by morphological characteristics, 16S rRNA sequence homology and molecular phylogeny. Maximum production of amylase by strain MG12 was found in pH 7.0 and 30°C with an incubation time of 48h. SDS-PAGE and characterization of enzyme showed 57.24 KDa with highest activity at pH 8 and 55°C. The Km and Vmax value for the enzyme activity was found using Line weaver-Burk plot.

References

  1. Asoodeh,A.Chamani, J., Lagzian, M. 2010. A novel thermostable, acidophilic α-amylase from a new thermophilic "Bacillus sp. Ferdowsicous" isolated from Ferdows hot mineral spring in Iran: Purification and biochemical characterization.International Journal of Biological Macromolecules.46.289-297.
  2. Al-Quadan.F., Akel.H. And NAtshi.R.2009. Characteristics of a novel highly thermostable and extremely thermophilic alkalitolerant amylase from a hyperthermophilic Bacillus strain HUTBS71.Online Journal of Biological Sciences.9.67-74
  3. Arikan.B. Highly thermostable,thermophilic,alkaline,SDS and chelator resistant amylase from a thermophilic Bacillus sp.isolate.A3-15.2008.Bioresourcr technology.99.3071-3076.
  4. Asger,M. Asad,M.J.Rahman,S.U and Legge,R.L.2007.A thermostable alpha amylase from a moderately thermophilic Bacillus subtilis strain for starch processing.Journal of food Engineering .79.950-955.
  5. Liu,B. Wang,Y and Zang,X.2006.Characterization of recombinant maltogenic amylase from deep sea thermophilic Bacillus sp.WPD 616.Enzyme and Microbial Technology.39.805-810.
  6. Hamilton.L.M., Kelly,C..T and Fogarty, W.M.1998. Carbohydrate Research, 314,251-257.
  7. Gupta.R., Gigras.P.,Mohapatra.H., Goswami..V.K and Chauhan.B(2003).Microbial alpha amylase:a biotechnological perspective .Process Biochemistry.38.1599-1616.
  8. Laemmli.U.K. 1970.Cleavage of structural proteins during the assembly of the head of bacteriophageT4.Nature 227.680-685.
  9. Lowry.O.H., Rosebrough.N.J., Farr.A.L.,Randall,R.J.1951.Protein measurement with the Folin-Phenol reagent.Jornal of Biological Chemistry.193.265-275.
  10. Miller.G.L. 1959.Use of dinitrosalicylic acid reagent for determination of reducing sugar.Analytical Chemistry.31.426-428.
  11. Morgan.F. and Priest.F.1981.Characterization of a thermostable alpha amylase from Bacillus licheniformis NCIB 6346.Journal of Applied Bacteriology.50.107-114.
  12. Seneath.P.H.A. Mair.N.S., Sharpe.E.M. Holt.J.G.1986.Bergeys Manual of Systematic Bacteriology,Ninth ed., Williams and Wilkins ,Baltimore,1986.
  13. Lineweaver.H.. and Burk.D.1934.Journal of American Chemical Society.56.658-666.
  14. Ivanova.V., Dobreva .E.,Emanuilova.E.1993.Purification and Characterization of of thermostable alpha amylase from Bacillus licheniformis. Journal of Biotechnology.28.277-289.
  15. Mc Tigue..M.A.,Kelly.T.C.,Doyel.M.E.,Forgatty.M.W.1995.The alkaline amylase of the alkaophilic Bacillus sp IMD 370.Enzyme and Microbial Technology.17.570-573.
  16. Mamo.G and Gesseses.A.1999. Purification and characterization of two starch digesting thermostable alpha amylase from thermophilic Bacillus.Enzyme and Microbial Technology.25.433-438.
  17. Huang.H.,Ridgwan.D.,Tingyue.GU.,MurrayMoo.Y.(2003).A segregated model forheterologus amylase production by Bacillus subtilis.Enzyme and Microbial Technology.37.407-413.
  18. Bernfeld, P., 1955. Amylases Alpha and Beta. In: Methods in Enzymology, Colowick, S.B. and N.O. Kaplan (Eds.). Academic Press, New York, pp: 149-153.
  19. Wanderely.K.J.,Torres.F.A.G.,Moraes.L.M.P.,Ulhoa.C.J.2004.Biochemical characterization of alpha amylase from the Yeast Cryptococcus flavus.FEMS Microbiolagy Letters.231.165-169.
  20. Kelly.C.T.,Bolton.D.J and Fogarty.W.M.1997.Biphasic production of alpha amylase of Bacillus flavothermus in batch fermentation. Biotechnology Letters.19.675-677.
  21. Hillier, P., D. Wase and A. Emery, 1996. Production of α-amylase by Bacillus amyloliquefaciens in batch and continuous culture using a defined synthetic medium. Biotechnol. Lett., 18: 795-800.
  22. Burhan.A.,Nisa.U.,Gokhan.C.,Ashabil.A.,Osmair.G.2003.Enzymatic properties of novel thermostable thermophilic alkaline and chelator resistant amylase from an alkaphilic Bacillus sp Isolate.ANT-6.Process Biochemistry.38.1397-1403.
  23. Najafi.M.F and Kembhavi.2005.One step purification and characterization of an extracellular alpha amylase from marine Vibrio sp. Enzyme and Mcrobial Technology.36. 535-539.
  24. Castro.G.A.,Ferrero.M.A.,Mendez.B.S and Sineriz.F.1993.Screening and selection of bacteria with high amylolytic activity. Acta Biotechnol.13.197-201.
  25. Van-Leeuwen.J.H.,Jin.B and Patel.B.999.Mycelial morphology and fungal protein production from starch processing waste water in submerged cultures of Aspergillus oryzae. Process Biochemistry.34.335-340.
  26. Pratima Bajpai, Pramod K. Bajpai.1989. High-temperature alkaline -amylase from Bacillus iicheniformis TCRDC-B13
  27. Castro.PML.,Hayter.P.M,Ison.A.P and Bull.A.T.1992.Application of statistical design to the optimization of culture medium for recombinant interferon gamma production by Chinese hamster ovary cells.Applied Microbiology and Biotechnology.38.84-90.
  28. Lin.L.L.,Chyau.C.C and Hsu.W.H.998.Production and properties of a raw starch degrading amylase from the thermophilic and alkaliphilic Bacillus sp.TS-23

IInternational Journal of Scientific Research in Computer Science, Engineering and Information Technology

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Published

2019-12-30

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Volume 4, Issue 9, November-December-2019

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Research Articles

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How to Cite

[1]
Dr. Asha B M, Dr. N Sakthivel, Anuradha Singh, Dr. H Ananda Vardhan, " Thermophilic and Alkalophilic Amylase from Strain Bacillus Marinus MG 12" International Journal of Scientific Research in Computer Science, Engineering and Information Technology(IJSRCSEIT), ISSN : 2456-3307, Volume 4, Issue 9, pp.23-29, November-December-2019.