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Production and partial purification of α-amylase from a novel isolate Streptomyces gulbargensis

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Extracellular amylase production by a newly isolated alkali-thermotolerant strain Streptomyces gulbargensis DAS 131 was optimized and characterized. The highest amylase production was achieved by growing S. gulbargensis DAS 131 in media with 1% starch. Strain exhibited maximal activity at pH 9.0 and 45°C and relatively stable in alkaline conditions (pH 11). Starch and peptone were found to be the good source of carbon and nitrogen with a yield of 2,216.6 and 2,156.1 U, respectively. Maltose and maltotriose were the main end products of starch hydrolysis, indicating α-amylase activity. SDS-PAGE analysis revealed a monomeric form with a molecular weight of 55 kDa.

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References

  1. Pandey A (1990) Improvement in solid-state fermentation for glucoamylase production. Biol Wastes 34:11–19. doi:10.1016/0269-7483(90)90140-N

    Article  CAS  Google Scholar 

  2. Rao MB, Tanksale AM, Ghatge MS, Deshpandae VV (1998) Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 62:597–635

    PubMed  CAS  Google Scholar 

  3. Pandey A, Nigam P, Soccol CR, Soccol VT, Singh D, Mohan R (2000) Advances in microbial amylases. Biotechnol Appl Biochem 31:135–152. doi:10.1042/BA19990073

    Article  PubMed  CAS  Google Scholar 

  4. Chandrasekarn S, Dhar SC (1987) Multiple protease from Streptomyces moderatus I. Isolation and purification of five extracellular proteases. Arch Biochem Biophys 257:395–401. doi:10.1016/0003-9861(87)90582-0

    Article  Google Scholar 

  5. James PDA, Iqbal M, Edwards C, Miller PGG (1991) Extracellular protease activity in antibiotic producing Streptomyces thermoviolaceus. Curr Microbiol 22:377–382. doi:10.1007/BF02092158

    Article  CAS  Google Scholar 

  6. Andrews L, Ward J (1988) Extracellular amylases from Streptomyces aureofaciens—purification and properties. Starch/Starke 30:338–341

    Google Scholar 

  7. Goldberg JD, Edwards C (1990) Purification and characterization of an extracellular amylase from a thermophilic Streptomyces. J Appl Bacteriol 69:712–717

    CAS  Google Scholar 

  8. Dastager SG, Wen-Jun Li, Dayanand A, Sulochana M, Shu-Kun Tang, Xin-Peng Tian, Xiao-Yang Zhi (2007) Streptomyces gulbargensis sp. nov., isolated from soil in Karnataka, India. Antonie Van Leeuwenhoek 91(2):99–104. doi:10.1007/s10482-006-9099-1

    Article  PubMed  Google Scholar 

  9. Goodfellow M (1989) Supergenic classification of actinomycetes. In: Williams ST, Sharpe ME, Holt JG (eds) Bergey’s manual of systematic bacteriology, vol 4. Williams and Wilkins, Baltimore, pp 2333–2339

    Google Scholar 

  10. Pridham TG, Anderson P, Foley C, Lindenfelser LA, Hesseltine CW, Benedict RG (1956) A selection of media for maintenance and taxonomic study of Streptomyces. Antibiot Annu 1:947–953

    Google Scholar 

  11. Hopwood DA, Bibb MJ, Chater KF, Kieser T, Gruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schirempe H (1985) Genetic manipulation of Streptomyces, a laboratory manual. John Innes Institute, Norwich, p 356

  12. Bernfeld P (1955) Amylases alpha and beta. In: Colowick SP, Kaplan ON (eds) Methods in enzymology. Academic Press, New York, pp 140–146

    Google Scholar 

  13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin-phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  14. Green AA, Hughens WL (1995) Protein fractionation on the basis of solubility in aqueous solution of salts and organic solvents. Methods Enzymol 1:67–90. doi:10.1016/0076-6879(55)01014-8

    Article  Google Scholar 

  15. Plumer DT (1978) An introduction to practical biochemistry. McGraw-Hill, London, pp 47–98

    Google Scholar 

  16. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of becteriophage T4. Nature 227:680–685. doi:10.1038/227680a0

    Article  PubMed  CAS  Google Scholar 

  17. Weber K, Osborn W (1969) The reliability of molecular weight determination by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 244:4406–4412

    PubMed  CAS  Google Scholar 

  18. Coronado MJ, Carmen V, Hofemeister J, Ventosa A, Nieto JJ (2000) Production and biochemical characterization of an α-amylase from the moderate halophile Halomonas meridianai. FEMS Microbiol Lett 183:67–71

    PubMed  CAS  Google Scholar 

  19. Takaya T, Sugimato Y, Wako K, Fuwa H (1979) Degradation of starch granules by alpha-amylase of Streptomyces praecox NA-273. Starch/Starke 31:205–208

    Article  CAS  Google Scholar 

  20. Fairbarn DA, Priest FG, Stark JR (1986) Extracellular amylase synthesis by Streptomyces limosus. Enzyme Microb Technol 8:89–92. doi:10.1016/0141-0229(86)90077-3

    Article  Google Scholar 

  21. Virolle MJ, Gagnat J (1994) Sequences involved in growth phase-dependent expression and glucose repression of a Streptomyces α-amylase gene. Microbiology 140:1059–1067

    Article  PubMed  CAS  Google Scholar 

  22. Onishi H, Sonoda K (1979) Purification and some properties of an extracellular amylase from a moderate halophile, Micrococcus halobius. Appl Environ Microbiol 38:616–620

    PubMed  CAS  Google Scholar 

  23. Egas MCV, Da Costa MS, Cowan DA, Pires EMV (1998) Extracellular α-amylase from Thermus filiformis Ork A2: purification and biochemical characterization. Extremophiles 2:23–32. doi:10.1007/s007920050039

    Article  PubMed  CAS  Google Scholar 

  24. Leveque E, Haye B, Belarbi A (2000) Cloning and expression of and α-amylase from the hyperthermophilic archaebacterium Thermococcus hydrothermalis and biochemical characterization of the recombinant enzymes. FEMS Microbiol Lett 186:67–71

    Article  PubMed  CAS  Google Scholar 

  25. Mijts BN, Patel BKC (2002) Cloning, sequencing and expression of and α-amylase gene, amy A, from the thermophilic halophilic Halothermothrix orenii and purification and biochemical characterization of the recombinant enzyme. Microbiology 148:2343–2349

    PubMed  CAS  Google Scholar 

  26. Zhu W, Dongmei C, Gugue C, Qian P, Ping S (2007) Purification and characterization of a thermostable protease from a newly isolated Geobacillus sp. YMTC 1049. Enzyme Microbiol Technol 40:1592–1597. doi:10.1016/j.enzmictec.2006.11.007

    Article  CAS  Google Scholar 

  27. De Azeredo LAI, Freire DMG, Soares RMA, Leite SGF, Coelho RRR (2004) Production and partial characterization of thermophilic proteases from Streptomyces sp isolated from Brazilian cerrado soil. Enzyme Microb Technol 34:354–358. doi:10.1016/j.enzmictec.2003.11.015

    Article  CAS  Google Scholar 

  28. Burhan A, Nisa U, Gökhan C, Ömer C, Ashabil A, Osman G (2003) Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. Isolate ANT–6. Process Biochem 38:1397–1403. doi:10.1016/S0032-9592(03)00037-2

    Article  CAS  Google Scholar 

  29. Horikoshi K (1999) Alkaliphiles: some applications of their products for biotechnology. Microbiol Mol Biol Rev 63:735–750

    PubMed  CAS  Google Scholar 

  30. Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B (2003) Microbial α-amylases: a biotechnological perspective. Process Biochem 1599–616. doi:10.1016/S0032-9592(03)00053-0

  31. Eva CMJ, Bernhardsdotter JD, Ng OK, Garriott ML, Pusey ML (2005) Enzymic properties of an alkaline chelator-resistant a-amylase from an alkaliphilic Bacillus sp. isolate L1711. Process Biochem 40:2401–2408. doi:10.1016/j.procbio.2004.09.016

    Article  CAS  Google Scholar 

  32. Igarashi K, Hatada Y, Hagihara H, Saeki K, Takaiwa M, Uemura T (1998) Enzymatic properties of a novel liquefying a-amylase from an alkaliphilic Bacillus isolate and entire nucleotide and amino acid sequence. Appl Environ Microbiol 64:3282–3289

    PubMed  CAS  Google Scholar 

  33. Mamo G, Gessesse A (1999) Purification and characterization of two raw-starch-digesting thermostable a-amylases from a thermophilic Bacillus. Enzyme Microb Technol 25:433–438. doi:10.1016/S0141-0229(99)00068-X

    Article  CAS  Google Scholar 

  34. Kim TU, Gu BG, Jeong JY, Byun SM, Shin YC (1996) Purification and characterization of a maltotetraose-forming alkaline a-amylase from an alkalophilic Bacillus strain GM8901. Appl Environ Microbiol 61:3105–3112

    Google Scholar 

  35. Hagihara H, Igarashi K, Hayashi Y, Endo K, Ikawa-Kitayama K, Ozaki K (2001) Novel a-amylase that is highly resistant to chelating reagents and chemical oxidants from the alkaliphilic Bacillus isolate KSM-K38. Appl Environ Microbiol 67:1744–1750. doi:10.1128/AEM.67.4.1744-1750.2001

    Article  PubMed  CAS  Google Scholar 

  36. Lin L-L, Chyau C-C, Hsu W-H (1996) Production and properties of a raw starch-degrading amylase from the thermophilic and alkaliphilic Bacillus sp. TS-23. Biotechnol Appl Biochem 28:61–68

    Google Scholar 

  37. Horikoshi K (1996) Alkaliphiles—from an industrial point of view. FEMS Microbiol Rev 18:259–270

    CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank CSIR Task force network programme on Exploration of India’s Rich Microbial Diversity (NWP 0006) for providing the financial support.

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Authors and Affiliations

  1. Biotechnology Division, National Institute of Interdisciplinary Science and Technology (CSIR), Industrial Estate, Pappanamcode, Thiruvananthapuram, 695019, India

    Dastager G. Syed & Ashok Pandey

  2. Department of Studies and Research in Microbiology, Gulbarga University, Gulbarga, 585 106, Karnataka, India

    Dayanand Agasar

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  1. Dastager G. Syed
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Correspondence to Ashok Pandey.

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Syed, D.G., Agasar, D. & Pandey, A. Production and partial purification of α-amylase from a novel isolate Streptomyces gulbargensis . J Ind Microbiol Biotechnol 36, 189–194 (2009). https://doi.org/10.1007/s10295-008-0484-9

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