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Isolation of Thermostable Alpha-amylase Producing Mesophilic Bacterium and Partial Characterization of the Enzyme

Year 2020, Volume: 3 Issue: 2, 146 - 153, 15.12.2020
https://doi.org/10.47495/okufbed.824607
https://izlik.org/JA24ZU24HD

Abstract

References

  • Referans1 Karademir G., Karademir B. Yem katkı maddesi olarak kullanılan biyoteknolojik ürünler, Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 2003; 43(1): 61-74, 2003.
  • Referans2 Nelson DL., Cox MM. Lehninger principles of biochemistry, Chapter 6, W.H. Freeman and Company, 4th Ed. New York, USA: 2004.
  • Referans3 Gümüşel F. Biyoteknoloji, genetik ve sağlık sektörü, Kocaeli Sanayii İçin Teknolojik Uzgörü Ortak Projesi 2002; 73-135.
  • Referans4 Wiseman A. The application of enzymes in industry. In: Horwood, E. (Ed.) Handbook of enzymes biotechnology. Chichester-UK 1987; 274-373.
  • Referans5 Gupta R., Gigras P., Mohapatra H., Goswami VK., Chauhan B. Microbial α-amylases: a biotechnological perspective, Process Biochemistry 2003; 38(11):1599-1616.
  • Referans6 Robyt JF. Enzymes and their action on starch. In: BeMiller, J., Whistler, R. (Eds.) Starch, chemistry and technology. Academic Press, New York: 2009; 237–292.
  • Referans7 Whitehurst RJ. Van Oort M. Enzymes in food tehcnology. In: Whitehurst, RJ., Van Oort, M. (Eds.), 2nd Ed. Wiley-Blackwell, Chichester, England: 2009.
  • Referans8 Leveque E., Janecek S., Haye B., Belarbi A. Thermophilic archaeal amylolytic enzymes-catalytic mechanism, substrate specificity and stability, Enyzme and Microbial Technology 2000; 26: 3-14.
  • Referans9 Asgher M., Asad MJ., Rahman SU., Legge RL. A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. Journal of Food Engineering 2007; 79(3): 950-955.
  • Referans10 Hewitt CJ., Solomons GL. The production of α-amylase (E.C.3.2.1.1.) by Bacillus amyloliquefaciens, in a complex and a totally defined synthetic culture medium. Journal of Industrial Microbiology 1996; 17(2): 96-99.
  • Referans11 Godfrey T., West S. Introduction to industrial enzymology. In: Godfrey, T., West, S. (Eds) New York, USA: Stockton Press 1996.
  • Referans12 Canoğulları S. Etlik piliç karma yemlerinde enzim kullanımı ve kullanım koşulları, Doktora Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana, Türkiye, 1999.
  • Referans13 Lennete EH., Ballows A., Hausler JW Jr., Shadomy JH. Manuel of clinical microbiology, American Society for Microbiology, Washington D.C., USA: 1985; 4:1149.
  • Referans14 Demirkan S. Production, purification, and characterization of α-amylase by Bacillus subtilis and its mutant derivates, Turkish Journal of Biology 2011; 35:705-712.
  • Referans15 Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar, Analytical Chemistry 1959; 426-428.
  • Referans16 Burhan AI., Nisa U., Gokhan C., Omer C., Ashabil A., Osman G. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6, Process Biochemistry 2003; 38(10):1397–1403.
  • Referans17 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4, Nature 1970; 227: 680-685.
  • Referans18 Ikeda T., Yamazaki H., Yamashita K., Shinke R. The tetracycline inducible expression of alpha-amylase in Bacillus subtilis. Journal of Fermentation and Bioengineering 1992; 74: 58-60.
  • Referans19 Lee SP., Morikawa M., Takagi M., Imanaka T. Cloning of the aapT gene and characterization of its product, α-amylase-pullulanase (AapT), from thermophilic and alkaliphilic Bacillus sp. strain XAL601, Applied and Environmenatl Microbiology 1994; 60(10): 3764-3773.
  • Referans20 Remize F. Spore-formin bacteria, The microbiological quality of food. In: Bevilacqua, A., Corbo, M.R., Sinigaglia, M. (Eds) Elsevier Ltd., Woodhead Publishing, Chennai, India, 2017.
  • Referans21 Harwood CR. Bacillus subtilis and its relatives: Molecular biological and industrial workhorses, Elsevier Science Publishers Ltd. U.K: 1992; 10:247-256.
  • Referans22 Vehmaanperä J. Development of Bacillus strains for industrial enzyme production by gene technology, Academic Dissertation in General Microbiology, PhD Thesis, Helsinki-Finland, 1990.
  • Referans23 Özcan BD. Studies on cloning of thermostable amylase genes, Yüksek Lisans Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana, Türkiye, 2005.
  • Referans24 Grootegoed JA., Lauwers AM., Heinen W. Separation and partial purification of extracellular amylase and protease from Bacillus caldolyticus, Archives of Microbiology 1973; 90:223.
  • Referans25 Bano S., Qader SAU., Aman A., Syed MN., Azhar A. Purification and characterization of novel α-Amylase from Bacillus subtilis KIBGE HAS, AAPS PharmSciTech 2011; 12(1): 255–261.
  • Referans26 Femi-Ola TO., Olowe BM. Characterization of alpha-amylase from Bacillus subtilis BS5 isolated from Amitermes evuncifer Silvestri, Research Journal of Microbiology 2011; 6(2):140-146.
  • Referans27 Ratanakhanokchai K., Kaneko J., Kamio Y., Izaki K. Purification and properties of a maltotetraose and maltotriose producing amylase from Chloroflexus aurantiacus, Applied and Environmental Microbiology 1992; 58:2490–2494.
  • Referans28 Matzke J., Schwermann B., Baker EP. Acidostable and acidophilic proteins: the example of the α-amylase from Alicyclobacillus acidocaldarius, Comparative Biochemistry and Physiology Part A: Physiology 1997; 118:475-479.
  • Referans29 Bozic N., Ruiz J., Lopez-Santin J., Vujcic V. Production and properties of the highly efficient raw starch digesting α-amylase from a Bacillus licheniformis ATCC 9945a, Biochemical Engineering Journal 2011; 53:203–209.
  • Referans30 Aguilar GJ., Morlon-Guyot B., Trejo-Aguilar C., Guyot JP. Purification and characterization of an extracellular α-amylase produced by Lactobacillus manihotivorans LMG 18010T an amylolytic lactic acid bacterium, Enzyme and Microbial Technology 2000; 27: 406–413.
  • Referans31 Liu XD., Xu Y. A novel raw starch digesting α-amylase from a newly isolated Bacillus sp. YX-1: Purification and characterization, Bioresource Technology 2008; 99: 4315-4320.

Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu

Year 2020, Volume: 3 Issue: 2, 146 - 153, 15.12.2020
https://doi.org/10.47495/okufbed.824607
https://izlik.org/JA24ZU24HD

Abstract

Bu çalışmada, Niğde ili Ulukışla ilçesi Çiftehan bölgesi sınırları içinde bulunan kaplıcadan alınan toprak örneklerinden α-amilaz aktivitesine sahip Bacillus izolasyonu yapılmıştır. İzolat bakteri, Bacillus sp. GA4 olarak isimlendirilmiştir. Bacillus sp. GA4 izolatına ait α-amilaz enziminin optimum pH ve sıcaklık değerleri sırasıyla 6.0-8.0 ve 50 ºC olarak bulunmuştur. Enzim 40 ºC’de 30 dk. ön inkübasyon sonrasında aktivitesini tamamen korumuştur. Enzim, 50 ºC ve 60 ºC’de 30 dk. ön inkübasyon sonrasında ise sırasıyla %21 ve %37’lik aktivite kaybına uğramıştır. MgCl2 ve FeSO4 enzim aktivitesini sırasıyla %13 ve %12 seviyelerinde artırırken, CuSO4, CaCl2 ve EDTA sırasıyla %81, %38 ve %33 oranlarında inhibe etmiştir. Bacillus sp. GA4 α-amilaz enziminin moleküler ağırlığı SDS-PAGE zimogram analizi ile yaklaşık olarak 55 kDa bulunmuştur. İzolatın CFX, CN, TE, RD, S, Amp ve P antibiyotiklerine karşı hassas, SH antibiyotiğine ise dirençli olduğu gözlenmiştir.

References

  • Referans1 Karademir G., Karademir B. Yem katkı maddesi olarak kullanılan biyoteknolojik ürünler, Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 2003; 43(1): 61-74, 2003.
  • Referans2 Nelson DL., Cox MM. Lehninger principles of biochemistry, Chapter 6, W.H. Freeman and Company, 4th Ed. New York, USA: 2004.
  • Referans3 Gümüşel F. Biyoteknoloji, genetik ve sağlık sektörü, Kocaeli Sanayii İçin Teknolojik Uzgörü Ortak Projesi 2002; 73-135.
  • Referans4 Wiseman A. The application of enzymes in industry. In: Horwood, E. (Ed.) Handbook of enzymes biotechnology. Chichester-UK 1987; 274-373.
  • Referans5 Gupta R., Gigras P., Mohapatra H., Goswami VK., Chauhan B. Microbial α-amylases: a biotechnological perspective, Process Biochemistry 2003; 38(11):1599-1616.
  • Referans6 Robyt JF. Enzymes and their action on starch. In: BeMiller, J., Whistler, R. (Eds.) Starch, chemistry and technology. Academic Press, New York: 2009; 237–292.
  • Referans7 Whitehurst RJ. Van Oort M. Enzymes in food tehcnology. In: Whitehurst, RJ., Van Oort, M. (Eds.), 2nd Ed. Wiley-Blackwell, Chichester, England: 2009.
  • Referans8 Leveque E., Janecek S., Haye B., Belarbi A. Thermophilic archaeal amylolytic enzymes-catalytic mechanism, substrate specificity and stability, Enyzme and Microbial Technology 2000; 26: 3-14.
  • Referans9 Asgher M., Asad MJ., Rahman SU., Legge RL. A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. Journal of Food Engineering 2007; 79(3): 950-955.
  • Referans10 Hewitt CJ., Solomons GL. The production of α-amylase (E.C.3.2.1.1.) by Bacillus amyloliquefaciens, in a complex and a totally defined synthetic culture medium. Journal of Industrial Microbiology 1996; 17(2): 96-99.
  • Referans11 Godfrey T., West S. Introduction to industrial enzymology. In: Godfrey, T., West, S. (Eds) New York, USA: Stockton Press 1996.
  • Referans12 Canoğulları S. Etlik piliç karma yemlerinde enzim kullanımı ve kullanım koşulları, Doktora Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana, Türkiye, 1999.
  • Referans13 Lennete EH., Ballows A., Hausler JW Jr., Shadomy JH. Manuel of clinical microbiology, American Society for Microbiology, Washington D.C., USA: 1985; 4:1149.
  • Referans14 Demirkan S. Production, purification, and characterization of α-amylase by Bacillus subtilis and its mutant derivates, Turkish Journal of Biology 2011; 35:705-712.
  • Referans15 Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar, Analytical Chemistry 1959; 426-428.
  • Referans16 Burhan AI., Nisa U., Gokhan C., Omer C., Ashabil A., Osman G. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6, Process Biochemistry 2003; 38(10):1397–1403.
  • Referans17 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4, Nature 1970; 227: 680-685.
  • Referans18 Ikeda T., Yamazaki H., Yamashita K., Shinke R. The tetracycline inducible expression of alpha-amylase in Bacillus subtilis. Journal of Fermentation and Bioengineering 1992; 74: 58-60.
  • Referans19 Lee SP., Morikawa M., Takagi M., Imanaka T. Cloning of the aapT gene and characterization of its product, α-amylase-pullulanase (AapT), from thermophilic and alkaliphilic Bacillus sp. strain XAL601, Applied and Environmenatl Microbiology 1994; 60(10): 3764-3773.
  • Referans20 Remize F. Spore-formin bacteria, The microbiological quality of food. In: Bevilacqua, A., Corbo, M.R., Sinigaglia, M. (Eds) Elsevier Ltd., Woodhead Publishing, Chennai, India, 2017.
  • Referans21 Harwood CR. Bacillus subtilis and its relatives: Molecular biological and industrial workhorses, Elsevier Science Publishers Ltd. U.K: 1992; 10:247-256.
  • Referans22 Vehmaanperä J. Development of Bacillus strains for industrial enzyme production by gene technology, Academic Dissertation in General Microbiology, PhD Thesis, Helsinki-Finland, 1990.
  • Referans23 Özcan BD. Studies on cloning of thermostable amylase genes, Yüksek Lisans Tezi, Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana, Türkiye, 2005.
  • Referans24 Grootegoed JA., Lauwers AM., Heinen W. Separation and partial purification of extracellular amylase and protease from Bacillus caldolyticus, Archives of Microbiology 1973; 90:223.
  • Referans25 Bano S., Qader SAU., Aman A., Syed MN., Azhar A. Purification and characterization of novel α-Amylase from Bacillus subtilis KIBGE HAS, AAPS PharmSciTech 2011; 12(1): 255–261.
  • Referans26 Femi-Ola TO., Olowe BM. Characterization of alpha-amylase from Bacillus subtilis BS5 isolated from Amitermes evuncifer Silvestri, Research Journal of Microbiology 2011; 6(2):140-146.
  • Referans27 Ratanakhanokchai K., Kaneko J., Kamio Y., Izaki K. Purification and properties of a maltotetraose and maltotriose producing amylase from Chloroflexus aurantiacus, Applied and Environmental Microbiology 1992; 58:2490–2494.
  • Referans28 Matzke J., Schwermann B., Baker EP. Acidostable and acidophilic proteins: the example of the α-amylase from Alicyclobacillus acidocaldarius, Comparative Biochemistry and Physiology Part A: Physiology 1997; 118:475-479.
  • Referans29 Bozic N., Ruiz J., Lopez-Santin J., Vujcic V. Production and properties of the highly efficient raw starch digesting α-amylase from a Bacillus licheniformis ATCC 9945a, Biochemical Engineering Journal 2011; 53:203–209.
  • Referans30 Aguilar GJ., Morlon-Guyot B., Trejo-Aguilar C., Guyot JP. Purification and characterization of an extracellular α-amylase produced by Lactobacillus manihotivorans LMG 18010T an amylolytic lactic acid bacterium, Enzyme and Microbial Technology 2000; 27: 406–413.
  • Referans31 Liu XD., Xu Y. A novel raw starch digesting α-amylase from a newly isolated Bacillus sp. YX-1: Purification and characterization, Bioresource Technology 2008; 99: 4315-4320.
There are 31 citations in total.

Details

Primary Language Turkish
Subjects Structural Biology
Journal Section Research Article
Authors

Gültekin Özdemir 0000-0002-4151-5399

B. Devrim Özcan 0000-0002-9198-656X

Submission Date November 11, 2020
Acceptance Date November 27, 2020
Publication Date December 15, 2020
DOI https://doi.org/10.47495/okufbed.824607
IZ https://izlik.org/JA24ZU24HD
Published in Issue Year 2020 Volume: 3 Issue: 2

Cite

APA Özdemir, G., & Özcan, B. D. (2020). Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 3(2), 146-153. https://doi.org/10.47495/okufbed.824607
AMA 1.Özdemir G, Özcan BD. Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu. Osmaniye Korkut Ata University Journal of The Institute of Science and Techno. 2020;3(2):146-153. doi:10.47495/okufbed.824607
Chicago Özdemir, Gültekin, and B. Devrim Özcan. 2020. “Sıcaklığa Dirençli Alfa-Amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu Ve Enzimin Kısmi Karakterizasyonu”. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi 3 (2): 146-53. https://doi.org/10.47495/okufbed.824607.
EndNote Özdemir G, Özcan BD (December 1, 2020) Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi 3 2 146–153.
IEEE [1]G. Özdemir and B. D. Özcan, “Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu”, Osmaniye Korkut Ata University Journal of The Institute of Science and Techno, vol. 3, no. 2, pp. 146–153, Dec. 2020, doi: 10.47495/okufbed.824607.
ISNAD Özdemir, Gültekin - Özcan, B. Devrim. “Sıcaklığa Dirençli Alfa-Amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu Ve Enzimin Kısmi Karakterizasyonu”. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi 3/2 (December 1, 2020): 146-153. https://doi.org/10.47495/okufbed.824607.
JAMA 1.Özdemir G, Özcan BD. Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu. Osmaniye Korkut Ata University Journal of The Institute of Science and Techno. 2020;3:146–153.
MLA Özdemir, Gültekin, and B. Devrim Özcan. “Sıcaklığa Dirençli Alfa-Amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu Ve Enzimin Kısmi Karakterizasyonu”. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, vol. 3, no. 2, Dec. 2020, pp. 146-53, doi:10.47495/okufbed.824607.
Vancouver 1.Gültekin Özdemir, B. Devrim Özcan. Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu. Osmaniye Korkut Ata University Journal of The Institute of Science and Techno. 2020 Dec. 1;3(2):146-53. doi:10.47495/okufbed.824607

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