Citrullus lanatus L. (Karpuz) ve Cucumis melo L. (Kavun) Kabuğu Kullanılarak Katı-Faz Fermentasyon Tekniği (SSF) ile Topraktan İzole Edilen Bacillus sp.’den Alkalin Serin Proteaz Eldesi

Abstract

Bu çalışmada, ekonomik önemi olmayan ve birikerek çevre kirliliğine yol açan karpuz ve kavun kabuğu katı substrat olarak kullanılarak Katı-Faz Fermentasyonu (SSF) yöntemi ile Bacillus sp.’den yüksek miktarda alkalin serin proteaz eldesi amaçlandı. Enzim aktivitesi üzerine pH, sıcaklık, metaller, azot ve karbon kaynakları etkisi incelendi. Yapılan inhibisyon çalışmasında PMSF ile inhibe olan enzimin alkalin serin proteaz olduğu tespit edildi. SSF yöntemi ekstrasellular enzimlerin düşük masrafla, hızlı bir şekilde, bol miktarda ve çevreye zarar vermeden üretilebildiği bir yöntem olduğu için kullanıldı.

Keywords

• Bacillus sp.,SSF,alkalin serin proteaz,kavun,karpuz

THE OBTAİNMENT OF ALKALİNE SERİNE PROTEASE FROM BACİLLUS SP. ISOLATED FROM SOLİD THROUGH THE SOLİD-STATE FERMENTATİON TECHNİQUE BY USİNG RİND OF THE CİTRULLUS LANATUS L. (WATERMELON) AND CUCUMİS MELO L. (MELON)

Abstract

In this study it was aimed to obtain serine alkaline protease in high amount from Bacillus sp. through solid-phase fermentation method (SSF) by using watermelon and melon rinds of no economic importance, and cause environment pollution by accumulation, as solid substrate. The effects of pH, temperature, metals, nitrogen and carbon sources were examined. In this inhibition study it was determined that the ezyme which is inhibited with PMSF (phenylmethylsulfonyl fluoride) was alkaline serine protease. SSF method was used since extracellular enzymes could be produced at low cost, in a quick way, in plenty and without giving any harm to the environment.

Keywords

• Bacillus sp.,SSF,alkaline serine protease,melon,watermelon

References

1. Agrawal, D., Patidar, P., Banerjee, T. & Patil, S. (2004) Production of alkaline protease by Penicillium sp. under SSF conditions and its application to soy protein hydrolysis. Process Biochemistry 39 977-981.
2. Aijun,Z., Hongzhang,C. & Zhou,L., (2005) Air pressure pulsation solid state production of alkaline protease by Bacillus pumilus 1.1625. Process Biochemistry, 40, 1547-1551.
3. Asther, M., Haon, M., Roussos, S., Record, E., Delattre, M., Lesagemeessen, L., Labat, M. & Asther, M. (2002) Feruloyl esterase from Aspergillus niger comparison of the production in solid state and submerged fermentation. Process Biochemistry 38, 685- 691.
4. Chelappan,S, Jasmin,C., Basheer,S.M., Elyas,K.K., Bhat,S.G., & Chandrasekaran,M. (2006) Production, purification and partial characterization of a novel protease from marine Engyodontium album BTMFS10 under solid state fermentation. Process Biochemistry 41, 956-961
5. Elibol,M & Antonio,R.M. (2004) Optimizing some factors affecting alkaline protease production by a marine bacterium Teredinobacter turnirae under solid state fermentation. Process Biochemistry Article in press.
6. Fujiwara,N. & Yamamoto,K. (1987) Production of alkaline protease in a low-cost medium by alkalophilic Bacillus sp. and properties of the enzyme. Fermentation Technology 65, 345-348.
7. Germano,S., Pandey, A., Osaku, C.A., Rocha, S.N. & Soccol, C.R. (2003) Characterization and stability of proteases from Penicillium sp. produced by solid state fermentation. Microbial Technology 32, 246-251.
8. Gupta, A., Roy, I., Patel, R.K., Singh, S.P., Khare, S.K. & Gupta, M.N. (2005) One-step purification and characterization of an alkaline protease from haloalkaliphilic Bacillus sp. Journal of Chromatography A, 1075, 103-108.

9. Johnvesly,B. & Naik, G.R. (2001) Studies on production of thermostable alkaline protease from thermophilic and alkalophilic Bacillus sp. JB-99 in a chemically defined medium. Process Biochemistry 37139-144.
10. Leighton,T.J., Doi,R.H., Warren,R.A.J. & Kelln, R.A. (1973) The relationship of serin protease activity to RNA polymerase modification and sporulation in Bacillus subtilis. Journal of Molecular Biology 22 76-103.
11. Müderriszade, A., Ensari, N.Y., Aguloglu, S. & Otludil, B. purification and characterization of alkaline proteinase from alkalophilic Bacillus sp. Applied Biochemistry and Microbiology, 37(2001) 574-577.
12. Pandey,A. (2003) Solid-state fermentation. Biochemical Engineering Journal 13 81-84.
13. Patel, R.K., Dodia, M.S., Joshi, R.H. & Singh, S.P. (2006) Purification and characterization of alkaline protease from a newly isolated haloalkaliphilic Bacillus sp. Process Biochemistry, 41, 2002-2009.
14. Puri, S., Beg, Q.K. & Gupta, R. (2002) Optimization of alkaline protease production from Bacillus sp. by response surface methodology. Current Microbiology 44 286-290.