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Topraktan İzole Edilen Bacillus Türlerinden α-Amilazın İzolasyonu ve Karakterizasyonu

Yıl 2019, Cilt: 6 Sayı: 3, 494 - 499, 23.07.2019
https://doi.org/10.30910/turkjans.595364

Öz

Toprak
mikroflorasının önemli bir kısmı Bacillus
cinsi bakterilerden oluşur. Bu bakterilerden elde edilen amilazların daha fazla
stabilite, yüksek verimlilik ve düşük üretim maliyetine sahip olma gibi
özellikleri vardı. Bundan dolayı topraktan izole edilen Bacillus cereus WYS01, Bacillus
thuringiensis
VITLW1, Bacillus
thuringiensis
ES2I3P, Bacillus cereus
TG16 mikroorganizmalardan α-amilazın izolasyonu ve karakterizasyonunu
gerçekleştirilmiştir. Yapılan araştırmada bütün bakterilerin maksimum enzim
üretiminin 36. saatte, pH 7.0 ve 40 ºC’de olduğu tespit edildi.
Mikroorganizmalar arasında spesifik aktivitesi en yüksek olan Bacillus cereus WYS01’tur. Enzim
aktivitesine sıcaklık ve pH gibi önemli etkisi olan parametrelere bakıldı. Bacillus thuringiensis VITLW1, Bacillus thuringiensis ES2I3P ve Bacillus cereus TG16 bakterilerinin 40
ºC de, Bacillus cereus WYS01’in ise
60 ºC de maksimum aktiviteyi gösterdiği tespit edildi. Bütün mikroorganizmalar
en fazla pH7.0 da α-amilaz aktivitesini gerçekleştirdi. Bu çalışmada
endüstriyel ve biyoteknolojik yönden önemi olan amilazın bakteriler tarafından
elde edildiği belirlendi.

Kaynakça

  • Aguloglu Fincan, S., Enez, B. 2014. Production, purification, and characterization of thermostable α-amylase from thermophilic Geobacillus stearothermophilus. Starch/Stärke, 66: 182-189.
  • Asgher, M., Asad, M.J., Rahman, S.U., Legge, R.L. 2007. A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. Journal Food Engineering, 79: 950-955.
  • Gupta, R., Gigras, P., Mohapatra, H., Goswami, V.K., Chauhan, B. 2003. Microbial - amylases: a biotechnological perspective. Process Biochemistry, 38: 1599-616.
  • Hamilton, L.M., Kelly, C.T., Fogarty, W.M. 1999. Production and properties of the raw starch digesting -amylase of Bacillus sp. IMD 435. Process Biochemistry, 35: 27-31.
  • Hmidet, N.,Bayoudh, A., Berrin, J.G., Kanoun, S., Juge, N., Nasri, M. 2008. Purification and biochemical characterization of a novel α-amylase from Bacillus licheniformis NH1 Cloning, nucleotide sequence and expression of amyN gene in Escherichia coli. Process Biochemistry, 43: 499-510.
  • John, R.J.D., Elangovan, N. 2013. Molecular identification of amylase producing Bacillus subtilis and detection of optimal conditions. Journal of Pharmacy Research, 6: 426-30.
  • Kikani, B.A., Singh, S.P. 2012. The stability and thermo dynamic parameters of a very thermostable and calcium-independent α–amylase from a newly isolated bacterium, Anoxybacillus beppuensis TSSC-1. Process Biochemistry, 47: 1791-1798.
  • Liu, X. D., Xu, Y. 2008. A novel raw starch digesting α-amylase from a newly isolated Bacillus sp. YX-1: Purification and characterization. Bioresource Technology, 99: 4315-4320.
  • Lowry, O. H., Roserbrough, N.J., Farr, A.L., Randall, R. 1951. Protein measure ment with folin phenol reagent. Journal of Biological Chemistry, 193: 265-275.
  • Miller, G.L. 1959. Use of dinitro salicylic acid reagent for determination of reducing sugars. Analytical Chemistry, 31: 426-428.
  • Najafi, M. F., Deobagkar. D., Deobagkar, D. 2005. Purification and characterization of an extracellular α-amylase from Bacillus subtilis AX20. Protein Expression and Purification, 41: 349-354.
  • Ortakaya, V., Aguloglu Fincan, S., Enez, B. 2017. α-Amylase from Bacillus simplex production, characterization and partial purification. Fresenius Environmental Bulletin. 26(7): 4446-4455.
  • Ozdemir, S, Aguloglu Fincan, S., Karakaya, A., Enez, B. 2018. A novel raw starch hydrolyzing thermostable α-amylase produced by newly isolated Bacillus mojavensis SO-10: Purification, characterization and usage in starch industries. Brazilian Archives of Biology and Technology, 61: e18160399.
  • Pandey, A., Nigam, P., Soccol, C.R., Soccol, V.T., Singh, D., Mohan, R. 2000. Advances in microbial amylases. Biotechnology and Applied Biochemistry, 31: 135-52.
  • Pranay, K., Padmadeo, S. R., Jha, V., Prasad, B. 2019. Screening and identification of amylase producing strains of Bacillus. Journal of Applied Biology Biotechnology, 1-6 DOI: 10.7324/JABB.2019.70103.
  • Saxena, R.K.,Dutt, K., Agarwal, L., Nayyar, P. 2007. A highly thermostable and alkaline amylase from a Bacillus sp. PN5. Bioresource Technology, 98: 260-265.
  • Shafiei, M.,Ziaee, A.A., Amoozegar, M.A. 2010. Purification and biochemical characterization of a novel SDS and surfactant stable, raw starch digesting and halophilic α- amylase from a moderately halophilicbacterium Nesterenkonia sp strain F. Process Biochemistry, 45: 694-699.
  • Souza, P.M., Magalhaes, P.O. 2010. Applications of microbial α-amylase in industry-a review. Brazilian Journal of Microbiology, 41: 850-61.
  • Topçal, F., Dığrak, M., Gündoğan, R. 2014. Identification of Bacillus Species Isolated from Soil and Determination of Bacteriocin Production. Adıyaman University Journal of Science, 4(2): 57-67.

Isolation and Characterization of α-Amylase from Bacillus Species Isolated from Soil

Yıl 2019, Cilt: 6 Sayı: 3, 494 - 499, 23.07.2019
https://doi.org/10.30910/turkjans.595364

Öz

An important part of the soil
microflora is made up of Bacillus bacteria.
Amylases obtained from these bacteria are known to have greater stability,
higher productivity and lower production costs. Therefore, isolation and
characterization of α-amylase from Bacillus
cereus
WYS01, Bacillus thuringiensis
VITLW1, Bacillus thuringiensis
ES2I3P, Bacillus cereus TG16
microorganisms isolated from soil was carried out. Inthestudy, the maximum
enzyme production of all bacteria was determined to be at pH7.0and 40 ºC at the
36th hour. Bacilluscereus WYS01 had
the highest specific activity among microorganisms. The parameters which have
important effects on the enzyme activity such as temperature and pH were
investigated. It was seen that Bacillus
thuringiensis
VITLW1, Bacillus
thuringiensis
ES2I3P and Bacillus
cereus
TG16 bacteria showed maximum activity at 40 ºC while this was 60 ºC
for Bacillus cereus WYS01. All
microorganisms carried out α-amylase activity at pH7.0. In this study, it was
determined that amylase, which is important for industrial and biotechnological
aspects, was obtained by bacteria.

Kaynakça

  • Aguloglu Fincan, S., Enez, B. 2014. Production, purification, and characterization of thermostable α-amylase from thermophilic Geobacillus stearothermophilus. Starch/Stärke, 66: 182-189.
  • Asgher, M., Asad, M.J., Rahman, S.U., Legge, R.L. 2007. A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. Journal Food Engineering, 79: 950-955.
  • Gupta, R., Gigras, P., Mohapatra, H., Goswami, V.K., Chauhan, B. 2003. Microbial - amylases: a biotechnological perspective. Process Biochemistry, 38: 1599-616.
  • Hamilton, L.M., Kelly, C.T., Fogarty, W.M. 1999. Production and properties of the raw starch digesting -amylase of Bacillus sp. IMD 435. Process Biochemistry, 35: 27-31.
  • Hmidet, N.,Bayoudh, A., Berrin, J.G., Kanoun, S., Juge, N., Nasri, M. 2008. Purification and biochemical characterization of a novel α-amylase from Bacillus licheniformis NH1 Cloning, nucleotide sequence and expression of amyN gene in Escherichia coli. Process Biochemistry, 43: 499-510.
  • John, R.J.D., Elangovan, N. 2013. Molecular identification of amylase producing Bacillus subtilis and detection of optimal conditions. Journal of Pharmacy Research, 6: 426-30.
  • Kikani, B.A., Singh, S.P. 2012. The stability and thermo dynamic parameters of a very thermostable and calcium-independent α–amylase from a newly isolated bacterium, Anoxybacillus beppuensis TSSC-1. Process Biochemistry, 47: 1791-1798.
  • Liu, X. D., Xu, Y. 2008. A novel raw starch digesting α-amylase from a newly isolated Bacillus sp. YX-1: Purification and characterization. Bioresource Technology, 99: 4315-4320.
  • Lowry, O. H., Roserbrough, N.J., Farr, A.L., Randall, R. 1951. Protein measure ment with folin phenol reagent. Journal of Biological Chemistry, 193: 265-275.
  • Miller, G.L. 1959. Use of dinitro salicylic acid reagent for determination of reducing sugars. Analytical Chemistry, 31: 426-428.
  • Najafi, M. F., Deobagkar. D., Deobagkar, D. 2005. Purification and characterization of an extracellular α-amylase from Bacillus subtilis AX20. Protein Expression and Purification, 41: 349-354.
  • Ortakaya, V., Aguloglu Fincan, S., Enez, B. 2017. α-Amylase from Bacillus simplex production, characterization and partial purification. Fresenius Environmental Bulletin. 26(7): 4446-4455.
  • Ozdemir, S, Aguloglu Fincan, S., Karakaya, A., Enez, B. 2018. A novel raw starch hydrolyzing thermostable α-amylase produced by newly isolated Bacillus mojavensis SO-10: Purification, characterization and usage in starch industries. Brazilian Archives of Biology and Technology, 61: e18160399.
  • Pandey, A., Nigam, P., Soccol, C.R., Soccol, V.T., Singh, D., Mohan, R. 2000. Advances in microbial amylases. Biotechnology and Applied Biochemistry, 31: 135-52.
  • Pranay, K., Padmadeo, S. R., Jha, V., Prasad, B. 2019. Screening and identification of amylase producing strains of Bacillus. Journal of Applied Biology Biotechnology, 1-6 DOI: 10.7324/JABB.2019.70103.
  • Saxena, R.K.,Dutt, K., Agarwal, L., Nayyar, P. 2007. A highly thermostable and alkaline amylase from a Bacillus sp. PN5. Bioresource Technology, 98: 260-265.
  • Shafiei, M.,Ziaee, A.A., Amoozegar, M.A. 2010. Purification and biochemical characterization of a novel SDS and surfactant stable, raw starch digesting and halophilic α- amylase from a moderately halophilicbacterium Nesterenkonia sp strain F. Process Biochemistry, 45: 694-699.
  • Souza, P.M., Magalhaes, P.O. 2010. Applications of microbial α-amylase in industry-a review. Brazilian Journal of Microbiology, 41: 850-61.
  • Topçal, F., Dığrak, M., Gündoğan, R. 2014. Identification of Bacillus Species Isolated from Soil and Determination of Bacteriocin Production. Adıyaman University Journal of Science, 4(2): 57-67.
Toplam 19 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Araştırma Makaleleri
Yazarlar

Sedat Kaya

Barış Enez

Yayımlanma Tarihi 23 Temmuz 2019
Gönderilme Tarihi 23 Mayıs 2019
Yayımlandığı Sayı Yıl 2019 Cilt: 6 Sayı: 3

Kaynak Göster

APA Kaya, S., & Enez, B. (2019). Topraktan İzole Edilen Bacillus Türlerinden α-Amilazın İzolasyonu ve Karakterizasyonu. Türk Tarım Ve Doğa Bilimleri Dergisi, 6(3), 494-499. https://doi.org/10.30910/turkjans.595364