α-AMYLASE PRODUCTION FROM Bacillus subtilis ATCC 6051 WITH SOLID STATE FERMENTATION (SSF)

Yıl 2013, Cilt: 1 Sayı: 2, 119 - 126, 01.12.2013

Sedat Kaya Yusuf Önen Fikret Uyar Nurullah Akcan

Öz

The usages of enzymes are used in biotechnology, industry, food, textil, paper, medicine and pharmaceutical. Enzymes obtained from microorganisms are produced by microorganisms have some advantages when compared with enzymes produced by plants or animals, they have considerably higher catalytic activity, they don’t from undesirable by-products, they are more stable and relatively cheap, and they can be obtained much quantity. The main goal of the present study is to realize the optimization parameters of the α-amylase obtained from Bacillus subtilis ATCC 6051 by solid phase fermentation method. The best incubation temperature, inkübation time and pH were determined for optimum enzyme activitiy. Enzyme showed optimum activity at 50 °C and ph 6.5. Amount of innoculum and initial moisture content suitable for enzyme production were found to be 30 % and 20 % respectively. The tap water was the best exraction medium

Anahtar Kelimeler

Solid State Fermentation (SSF), Bacillus subtilis, α-Amylase, Rice husk

KATI FAZ FERMENTASYONU (KSF) TEKNİĞİ ile Bacillus subtilis ATCC 6051’den

Öz

Enzimler genelde biyoteknolojide, endüstride, yiyecek, tekstil, kağıt, tıp ve eczacılık gibi alanlarda kullanılmaktadır. Enzimler; bitkisel, hayvansal kaynaklardan ve mikroorganizmalardan elde edilmektedir. Mikroorganizma kaynaklı enzimlerin bitkisel veya hayvansal kaynaklı enzimlere göre katalitik aktivitelerinin çok yüksek olmaları, istenmeyen yan ürün oluşturmamaları, daha stabil ve ucuz olmaları, kültür ortamında kolay çoğalmaları, fazla miktarda elde edilebilmeleri gibi avantajları vardır. Bu çalışmadaki amacımız Katı Faz Fermentasyon yöntemi ile Bacillus subtilis ATCC 6051’den α-amilaz üretimi ile ilgili parametrelerin optimizasyonu ile ilgili çalışmalar yapmaktır. Enzim üretimi için en iyi inkübasyon sıcaklığı, inkübasyon süresi ve pH’sı belirlendi. Enzimin optimum aktivite gösterdiği sıcaklık ve pH sırasıyla 50 oC ve 6.5 olarak tespit edilmiştir. Enzim için uygun inokülüm (ekim) miktarı % 30, başlangıç nem miktarı % 20 olarak tespit edilmiştir. Enzim için en iyi ekstraksiyon medyumu çeşme suyu olarak tespit edilmiştir.

Anahtar Kelimeler

Katı faz fermentasyonu (SSF), Bacillus subtilis, α-Amilaz, Pirinç kabuğu

Kaynakça

• Baysal, Z., Uyar, F., Doğru M., Alkan, H.. Production of Extracellular Alkaline a-Amylase by
• Solid State Fermentation with a Newly Isolated Bacillus sp. Biochemistry&Biotechnology, 38,
• 184–190, 2008.
• Bernfeld, P.. Enzymes carbohydrate metabolism, In Metods In Enzymology Academic Press,
• 17, 149-158, 1955.
• Gangadharan, D. K., Sivaramakrishnan, S., Nampoothiri, K. M., Soccol,C. R. Pandey A: α-
• Amylases from microbial sources an Overview on recent developments. Food Technol Biotechnol, 44, 173-184, 2006.
• Gupta, R., Gigras. P., Mohapatra, H., Goswami V.K., Chauhan, B. 2003. Microbial α-amylases: a biotechnological perspective. Process Biochem. 38, 1599-1616, 2006.
• Hashemi, M., Razavi, S.H., Shojaosadati, S.A., Mousavi, S.M., Khajeh, K., Safari.M. Development of a solid-state fermentation process for production of an alphaamylase with potentially interesting properties Journal of Bioscience and Bioengineering. 3, 333–337, 2010.
• Hashemi, M., Mousavi, S.M., Razavi, S.H., Shojaosadati, S.A. Mathematical modeling of
• biomass and amylase production kinetics by Bacillus sp. İn solid-state fermentation based
• on solid dry weight variation Biochemical Engineering Journal, 53, 159–164, 2011.
• Kandra, L. α-Amylases of medical and industrial importance; Journal of Moleculer Structure
• (Teochem) 666-667 p. 487-498, 2003.
• Kar, S., Data,T.K., Ray, R.C. Optimization of Thermostable α- Amylase Production by Streptomyces erumpens MTCC 7317 in Solid-state Fermentation Using Cassava Fibrous Residue, Brazılıan Archıeves of Bıolıgy and Technology an İnternational journal 53, 301-309, 2010.
• Kıran, Ö.E., Çömlekçioğlu,U., Dostbil, N., Bazı mikrobiyal enzimler ve endüstrideki kullanım
• alanları, KSÜ Fen ve Mühendislik Dergisi, 9, 1, 2006.
• Krisha, C., Chandrasekaran M., Banana Waste As Substrate For α-Amylase Production By
• Bacillus Subtilis Cbtk 106 Under Solid-State Fermentation ; Microbial Biotechnology, 46:
• 106-111, 1996.
• Kunamneni, A., Permaul, K., Singh, S. Amylase production in solid state fermentation by the
• thermophilic fungus Thermomyces lanuginosus, Journal of Bioscience and Bioengineering,
• 100, 168-171, 2005.
• Lowry, O.H., Rosebrough, N.J., Farr, A.L., Protein measurement with the folin phenol reagent, J. Biol. Chem. 193, 265-275, 1951.
• Michelin, M., Silva, T. M., Benassi, V. M., Peixoto- Nogueira, S. M., Moraes, L.A., Leão, J.
• M., Jorge, J. A., Terenzi, H. F., Polizeli, M. L. Purification and characterization of a thermostable a-amylase produced by the fungus Paecilomyces variotii, Carbohydrate Research ,345, 2348–2353, 2010.
• Pandey, A., Selvakumar, P., Soccol, C.R., Nigam, P. Solid state fermentation for the production of industrial enzymes, Current Science, 77,149-162 The application of enzymes in
• industry, 274-373, 1999.
• Pandey, A., Soccol, C.R., Mitchell, D. New developments in solid state fermentation:I –Bioprocesses and products. Process Biochem 35, 1153–1169, 2000.
• Pandey A, C.R., Soccol, J.A.R., Leon, Nigam, P. Solid-state fermentation in biotechnology. New Delhi: Asiatech Publishers Inc. 2004.
• Pandey, A. Ricardo, C. Larroche, C. Current Developments in Solid-state Fermentation
• Springer Science Business Media, 144, 1–22, 2008.
• Rıaz, N., Haq , H.K., Qadder, M.A. Characterization of α-Amylase by Bacillus subtilis International Journal of Agriculture & Biology, 3–249–252, 2003.
• Serin, B. Katı faz tekniğiyle (solid state fermentation; SSF) tekniğiyle Bacillus circulans‘tan
• α-amilaz ve β-galaktosidaz üretimi, Yüksek Lisans Tezi, Dicle Üniversitesi Fen Bilimleri Enstitüsü, 47s, 2009.
• Shukla, J., Kar, R. Potato peel as a solid state substrate for thermostable aamylase production by thermophilic Bacillus isolates World Journal of Microbiology & Biotechnology, 22, 417–422, 2006.
• Singhaniaa, R. R., Patel, A. K., Soccol, C. R., Pandey, A. Recent advances in solid-state fermentation Biochemical Engineering Journal, 44, 13–18, 2009.
• Sodhi, H. K., Sharma, K., Gupta, J. K., Soni, S. K. Production of a thermostable α-amylase
• from Bacillus PS-7 by solid state fermentation and its synergitic use in the hydrolysis of malt
• starch for alcohol production, Process Biochemistry, 40, 525-534, 2005.
• Taniguchi, H., Honda, Y. Ishikawa Prefectural University, Nonoichi, Elsevier Inc. All rights reserved. Defining Statement,159-179, 2009.
• Vishnu, C., Navenna, B. J., Altaf, MD., Venkateshwear, M., Reddy, G., Amylopullulanase A
• novel enzyme of L. amylophilus GV6 in direct fermentation of starch to L(+) lactic acid, Enzyme Microb Technol, 38, 545–550, 2006.
• Xu, H., Sun, L., Zhao, D., Zhang, B., Shi, Y., Wu, Y., Production of α- amylase by Aspergillus oryzae AS-3951 in solid state fermentation using spent brewing grains as Substrate, Journal of the Science of Food and Agriculture, 88, 529–535, 2008.
• Xu, Y. A., Liu, X.D. novel raw starch digesting α-amylase from a newly isolated Bacillus sp.
• YX-1: purification and characterization, Bioresource Technology, 99, 4315-4320, 2008