Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil

QAZI, Javed Iqbal; BAIG, Shahjahan; SYED, Qurat Ul Ain; Nadeem, Muhammad

Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil

Muhammad NADEEM, (Biotechnology and Food Research Center, PCSIR Laboratuvarı, Pakistan)

Javed Iqbal QAZI, (Department, of Zoology, Univercity of the Punjab, Lahore, Pakistan)

Shahjahan BAIG, (Biotechnology and Food Research Center, PCSIR Laboratuvarı, Pakistan)

Qurat Ul Ain SYED (Biotechnology and Food Research Center, PCSIR Laboratuvarı, Pakistan)

Türk Biyokimya Dergisi

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Yıl: 2007 Cilt: 32 Sayı: 4 Sayfa Aralığı: 171 - 177 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil

Öz:

Çürümekte olan organik toprağın üst yüzeyinden (0–5 cm) seksen bakteriyel suş izole edilmiştir. Otuz altı izolat (% 47.50) az yağlı süt agarında (pH 10) belirgin berraklaşma alanı göstermiştir. Bu suşlar karakterize edilmiş ve tanımlanmıştır. Alkalophilic izolatlardan otuz iki tanesi (% 40) Bacillus genomuna ait olduğu saptanmıştır. Bacillus lichniformis N-2 olarak belirlenen suşdan elde edilen maksimum enzim aktivitesi pH 10 ve 37 oC’de 123.29 ± 1.61 PU/ml’dir. Farklı sıcaklıklar, pH, metal iyonları, inhibitörler ve proteaz aktivitesi gibi bazı temel özelliklerin enzim aktivitesine etkisi de çalışılmıştır. Maksimum aktivite pH 11, 60 oC’da elde edilmiştir. Ca+2 ve Mg+2 iyonlarının göreceli enzim aktivitesini % 132 ve 118 oranlarında artırdığı gözlenmiştir. Buna rağmen proteaz aktivitesinin fenilmetilsulfonilflorid ile tamamen inhibe edilmiş olması bu enzimin bir serine proteaz olduğunu göstermektedir. Sonuçlarımız B. lichniformis-N2 tarafından sentezlenen enzimin geniş ısı ve pH aralıklarında aktif olduğunu göstermiştir. Bu karakteristikler sebebi ile deri ve deterjan endüstrilerinde kullanıma uygundur.

Anahtar Kelime:

Konular: Biyokimya ve Moleküler Biyoloji

Çürümekte olan organik topraktan izole edilen, bacillus lichniformis N-2 tarafından sentezlenen ve ticari önem taşıyan alkalen proteaz üzerine çalışmalar

Öz:

Eighty bacterial strains were isolated from upper layer (0-5 cm) of decaying organic soil sample. Thirty eight isolates (47.50 %) exhibited the prominent zones of clearance on skim milk agar medium at pH 10. These isolates were then characterized and identified. Thirty two (40 %) of the alkalophylic isolates belonged to the genus Bacillus. Maximum enzyme activity (123.29 ± 1.61 PU/ ml) was found in strain designated as Bacillus lichniformis N-2 at pH 10 and temperature 37 oC. Some fundamental properties like effects of different temperature, pH, metal ions and inhibitors on protease activity were also studied. Maximum activity was obtained at pH 11 at 60 oC. Ca+2 and Mg+2 ions were found to enhance the relative enzyme activity up to 132 and 118 % respectively. However, the activity of protease was completely inhibited by phenylmethylsulfonylflouride that showed its serine nature. The results indicated that enzyme produced by B. lichniformis-N2 is active within broad ranges of temperature and pH. These characteristics render its potential use in leather and detergent industries.

Anahtar Kelime:

Konular: Biyokimya ve Moleküler Biyoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

[1] Horikoshi K. (1999) Alkaliphiles: Some applications of their products for biotechnology. Microbiol. Mol Biol Rev. 63 (4): 735-750.
[2] Dermirijan DC, Moris VF, Cassidy CS. (2001) Enzymes from extremophilles. Cur Opin Chem Biol. 5 (2): 144-151.
[3] Nilegaonkar SS, Kanekar PP, Sarnaik SS, Kelkar AS. (2002) Production, isolation, and characterization of extra-cellular protease of an alkaliphillic strain of Arthrobacter ramosus, MCM B-351 isolated from the alkaline lake of Lonar, India. World J Microbiol Biotechnol. 18 (8): 785-789.
[4] Kanekar PP, Nilegaonkar SS, Sarnaik SS, Kelkar AS. (2002) Optimization of proteases activity of alkaliphilic bacteria isolated from an alkaline lake in India. Biores Technol. 85 (1): 87-93.
[5] Zeynep U, Metin D. (2002) Alkaliphilic microorganisms and habitats. Turkish J Biol. 26 (3): 181-191.
[6] Denizci AA, Kazan D, Abeln ECA, Erarslan A. (2004) Newly isolated Bacillus clausii GMBAE 42: an alkaline protease producer capable to grow under highly alkaline conditions. J Appl Microbiol. 96 (2): 320-327.
[7] Naidu KSB, Devi KL. (2005) Optimization of thermostable alkaline proteases production from species of Bacillus using rice bran. African J Biotechnol. 4 (7): 724-726.
[8] Gupta R, Beg QK, Lorenz L. (2002) Bacterial alkaline proteases: Molecular approaches and industrial applications. Appl Microbiol Biotechnol. 59 (1):15-32.
[9] Kumar CG, Takagi H. (1999) Microbial alkaline proteases: from a bio-industrial point of view. Biotechnol Adv. 17 (7): 561-594.
[10] Holt JG, Krieg NR, Sneath PHA, Stately JT, William ST. (1994) Bergey’s Manual of Determinative Bacteriology, 9th Ed., Williams and Wilkins, Baltimore, Maryland, USA, pp. 559.
[11] Betty AF, Daniel FS, Alice, SW. (2002) Bacillus and similar organisms. In: Diagnostic Microbiology, , 11th Edition, Published by Mosby Inc. 11830 West line Industrial Drive, St. Louis, Missouri 63146, USA, pp 317-324.
[12] Yang SS, Huang CI. (1994) Proteases production by amylolytic fungi in solid state fermentation. J Chinese Agri Chem Soc. 32 (6): 589-601.
[13] Lowry OH, Roserbrough NJ, Farr AL, Randall, R. (1951) Protein measurement with Folin Phenol Reagent. J Biol Chem. 193 (1): 265-275.
[14] Jeroen H, Bram V, Niall AL, Balcaen A, Marina RD, Andreas S. Paul, D. V. (2004) Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataveinsis sp. nov, and Bacillus drentensis sp. nov., from the Drentse A grasslands. Int Syst Evolution Microbiol. 54 (1): 47-57.
[15] Belma A, Zehra NY, Yavuz B. (2002) Determination of PHB growth quantities of certain Bacillus species isolated form soil. Turkish Elect J Biotechnol Special issue: 24-30.
[16] Bach HJ, Munch JC. (2000) Identification of bacterial sources of soil peptidases. Biol. Fertil. Soils. 34 (3-4): 219-224.
[17] Kumar CG, Malik RK, Tiwari MP. (1998) Novel enzyme based detergents: an Indian perspective. Curr Sci. 75 (12): 1312-1318.
[18] Pastor MD, Lorda GS, Balatti A. (2001) Proteases production using Bacillus subtilis- 3411 and amaranth seed meal medium at different aeration rate. Brazilian J Microbiol. 32 (1): 6-9.
[19] Moradian F, Khajeh K, Naderi-Manesh H, Ahmadvand R, Sajedi RH, Sadeghizadeh M. (2006) Thiol-dependeant serine alkaline proteases from Bacillus sp. HR-08 and KR-8102: isolation, production and characterization. Appl Biochem Biotechnol. 134 (1): 77-87
[20] Joo HS, Kumar CG, Park GC, Palk SR, Chang CS. (2003) Oxidant and SDS-stable alkaline protease from Bacillus clausii 1-52: production and some properties. J Appl Microbiol. 95 (2): 267-272.
[21] Horton HH, Moran LA, Raymond SO, Rawn JD, Scrimgeour KG. (1996) Properties of enzymes. In: Principles of Biochemistry. 2nd edition, printed by prentice-hall ınternational, Inc. USA, p. 119-146.
[22] Jeong YH, Wei CI, Preston JF, Marshall MR. (2000) Purification and characterization of proteases from hepatopancreas of crawfish (Procambarus clarkii). J Food Biochem. 24 (4): 311-332.
[23] Adinarayana K, Ellaiah P, Prasad DS. (2003) Purification and characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. Pharma Sci Technol. 4 (4): 1-9.
[24] Siriporn Y, Alissara R, Masaki Y. (2006) Purification and characterization of alkaline protease from Bacillus megatarium isolated from thai fish Sauce fermentation process. Sci Asia 32: 377-388.
[25] Johnvesly B, Naik GR. (2001) Studies of production of thermostable alkaline protease from thermophilic and alklaliphilic Bacillus sp. JB-99 in a chemical defined medium. Process Biochem. 37 (2): 139-144.
[26] Donaghy JA, Mckay AM. (1993) Production and properties of alkaline proteases by Aureobasidium pullulans. J Appl Bacteriol. 74 (6): 662-666.

APA	Nadeem M, QAZI J, BAIG S, SYED Q (2007). Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. , 171 - 177.
Chicago	Nadeem Muhammad,QAZI Javed Iqbal,BAIG Shahjahan,SYED Qurat Ul Ain Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. (2007): 171 - 177.
MLA	Nadeem Muhammad,QAZI Javed Iqbal,BAIG Shahjahan,SYED Qurat Ul Ain Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. , 2007, ss.171 - 177.
AMA	Nadeem M,QAZI J,BAIG S,SYED Q Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. . 2007; 171 - 177.
Vancouver	Nadeem M,QAZI J,BAIG S,SYED Q Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. . 2007; 171 - 177.
IEEE	Nadeem M,QAZI J,BAIG S,SYED Q "Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil." , ss.171 - 177, 2007.
ISNAD	Nadeem, Muhammad vd. "Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil". (2007), 171-177.

APA	Nadeem M, QAZI J, BAIG S, SYED Q (2007). Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. Türk Biyokimya Dergisi, 32(4), 171 - 177.
Chicago	Nadeem Muhammad,QAZI Javed Iqbal,BAIG Shahjahan,SYED Qurat Ul Ain Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. Türk Biyokimya Dergisi 32, no.4 (2007): 171 - 177.
MLA	Nadeem Muhammad,QAZI Javed Iqbal,BAIG Shahjahan,SYED Qurat Ul Ain Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. Türk Biyokimya Dergisi, vol.32, no.4, 2007, ss.171 - 177.
AMA	Nadeem M,QAZI J,BAIG S,SYED Q Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. Türk Biyokimya Dergisi. 2007; 32(4): 171 - 177.
Vancouver	Nadeem M,QAZI J,BAIG S,SYED Q Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil. Türk Biyokimya Dergisi. 2007; 32(4): 171 - 177.
IEEE	Nadeem M,QAZI J,BAIG S,SYED Q "Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil." Türk Biyokimya Dergisi, 32, ss.171 - 177, 2007.
ISNAD	Nadeem, Muhammad vd. "Studies on commercially important alkaline protease from bacillus lichniformis N-2 isolated from decaying organic soil". Türk Biyokimya Dergisi 32/4 (2007), 171-177.