Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil

ÜLKER, Serdar; Çolak, Ahmet; Özel, Arzu; Alpay Karaoğlu, Şengül

Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil

Serdar ÜLKER, (Rize Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Rize, Türkiye)

Arzu ÖZEL, (Rize Üniversitesi, Fen Edebiyat Fakültesi Kimya Bölümü, Rize, Türkiye)

Ahmet ÇOLAK, (Karadeniz Teknik Üniversitesi, Fen Edebiyat Fakültesi, Kimya Bölümü, Trabzon, Türkiye)

Şengül Alpay KARAOĞLU (Rize Üniversitesi, Fen Edebiyat Fakültesi Kimya Bölümü, Rize, Türkiye)

Turkish Journal of Biology

1 0

Yıl: 2011 Cilt: 35 Sayı: 5 Sayfa Aralığı: 543 - 550 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil

Öz:

Bu çalışma, Trichoderma harzianum lipazının karakterizasyonu hakkında ilk rapordur. Yeni bir suş olan Trichoderma harzianum IDM14D topraktan izole edildi. İzole edilen bu suş, lipaz üretimi için çalkalamalı kültürde 30 °C’de 7 gün inkübe edilmiştir. Lipaz üretimi için en iyi karbon kaynağı glukoz, en iyi azot kaynağının pepton olduğu belirlendi. Maksimum biyokütle üretimi 7 gün sonunda 1,25 g/L olarak belirlendi. Enzimin en iyi aktivite gösterdiği pH 8,5, sıcaklık 40 °C olarak belirlendi. T. harzianum lipazı pH 8,0-10,0 aralığında 40 °C’de 60 dakika kararlılığını koruduğu belirlendi. Ca2+ ve Mn2+ iyonlarının lipaz aktivitesini artırdığı, fakat diğer metal iyonlarının enzim aktivitesini etkilemediği gözlendi. p-nitrofenil butirat hidrolizi ile ölçülen ham enzimin Km ve Vmaks değerleri sırasıyla 7,15 mM ve 7,067 mM/dk olarak belirlendi.

Anahtar Kelime: toprak Trichoderma harzianum triasilgliserol lipaz enzim aktivitesi lipaz saflaştırma hücre dışı proteinler karakterizasyon izolasyon

Konular: Biyoloji

Topraktan izole edilen Trichoderma harzianum hücre dışı lipazının izolasyonu, üretimi ve karakterizasyonu

Öz:

This is the first report about the characterization of Trichoderma harzianum lipase. A novel strain of Trichoderma harzianum IDM14D was isolated from soil. The isolated strain was cultivated for lipase production in shake fl asks at 30 °C for 7 days. For lipase production, it was determined that the best carbon source was glucose and the best nitrogen source was peptone. Maximum biomass was produced at a concentration of 1.25 g/L, in 7 days. The optimum pH and temperature for activity of the enzyme were 8.5 and 40 °C, respectively. The lipase was stable at a pH range of 8.0-10.0 and at 40 °C for 60 min. Ca2+ and Mn2+ enhanced lipase activity but it was determined that other metallic ions did not affect the enzyme activity. The Km and Vmax values of the crude enzyme for p-nitrophenyl butyrate hydrolysis were found to be 7.15 mM and 7.067 mM/min, respectively.

Anahtar Kelime: triacylglycerol lipase enzyme activity lipase purification extracellular proteins characterization isolation soil Trichoderma harzianum

Konular: Biyoloji

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

1. Maragoni AG. Candida and Pseudomonas lipase catalyzed hydrolysis of butter oil in the absence of organic solvents. J Food Sci 59: 1096-9, 1994.
2. Marek A, Bednarski W. Some factors aff ecting lipase production by yeasts and fi lamentous fungi. Biotechnol Lett 18: 1155-1160, 1996.
3. Mahadik ND, Puntambekar US, Bastawde KB et al. Production of acidic lipase by Aspergillus niger in solid state fermentation. Process Biochem 38: 715-721, 2002.
4. Pandey A. Solid-state fermentation. Biochem Eng J 13: 81-84, 2003.
5. Freire DMG, Teles EMF, Bon EPS et al. Lipase production by Penicillium restrictum in a bench-scale fermenter: Eff ect of carbon and nitrogen nutrition, agitation, and aeration. Appl Biochem Biotech 63: 409-421, 1997.
6. Gotor V. Non-conventional hydrolase chemistry; amide and carbamate bond formation catalyzed by lipases. Bioorg Med Chem 7: 2189-2197, 1999.
7. Saxena RK, Ghosh PK, Gupta R et al. Microbial lipases: Potential biocatalysts for the future industry. Curr Sci India 77: 101-115, 1999.
8. Shimada Y, Hirota Y, Baba T et al. Enzymatic synthesis of steryl esters of polyunsaturated fatty acids. J Am Chem Soc 76: 713- 716, 1999.
9. Th eil F. Lipase supported synthesis of biologically active compounds. Chem Rev 95: 2203-2227, 1995.
10. Godtfredsen SE. Microbial lipases. In: Fogarty WM, Kelly ET (eds). Microbial enzymes and biotechnology. Elsevier Applied Science Publishers. Amsterdam; 1990: pp. 255-274.
11. Ghosh PK, Saxena RK, Gupta R et al. Microbial lipases: production and applications. Sci Prog 79: 119-157, 1996.
12. Rubin B, Dennis EA, (eds), Methods in Enzymology: Lipases, Part B: Enzyme Characterization and Utilization (Vol. 286), Academic Press, Maryland Heights; 1997: pp. 509-519.
13. Castilho LR, Polato CMS, Baruque EA et al. Economic analysis of lipase production by Penicillium restrictum in solid-state and submerged fermentations. Biochem Eng J 4: 239-247, 2000.
14. Leal MCMR, Cammarota MC, Freire DMG et al. Hydrolytic enzymes as coadjuvants in the anaerobic treatment of dairy waste waters. Braz J Chem Eng 19: 175-180, 2002.
15. De Marco JL, Valadares-Inglis MC, Felix CR. Production of hydrolytic enzymes by Trichoderma isolates with antagonistic activity against Crinipellis perniciosa, the casual agent of witches’ broom of cocoa. Braz J Microbiol 34: 33-38, 2003.
16. Bayözen A, Yıldız A. Determination of mycorrhizae interactions and pathogenicity of Rhizoctonia solani Kühn isolated from strawberry and Xanthium strumarium. Turk J Biol 33: 53-57, 2009.
17. Topal Ş, Pembeci C, Borcaklı M. Türkiye’nin tarımsal mikofl orasının endüstriyel öneme sahip bazı enzimatik aktivitelerinin incelenmesi-I: Amilaz, proteaz, lipaz. Turk J Biol 24: 79-93, 2000.
18. Karaoglu SA, Ulker S. Isolation, identifi cation and seasonal distribution of soilborne fungi in tea growing areas of İyidereİkizdere vicinity (Rize-Turkey). J Basic Microb 46: 208-218, 2006.
19. Samson RA, Hoekstra ES. Introduction to food and airborne fungi, American Society for Microbiology, Centraalbureau voor Schimmelcultures (CBS), 2002.
20. Cihangir N, Sarikaya E. Investigation of lipase production by a new isolate of Aspergillus sp. World J Microb Biot 20: 193-197, 2004.
21. Pimentel MCB, Krieger N, Coelho LCCB et al. Lipase from a Brazilian strain of Penicillium citrinum. Appl Biochem Biotechnol 177: 59-74, 1994.
22. Lee D, Kok Y, Kim K et al. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1. FEMS Microbiol Lett 179: 393-400, 1999.
23. Lowry OH, Rosebrough NJ, Farr AL et al. Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275, 1951.
24. Musidlowska A, Lange S, Bornscheuer UT. Via overexpression in the yeast Pichia pastoris to enhanced enantioselectivity: New aspects in the application of pig liver esterase. Angew Chem Int Ed Engl 40: 2851-2853, 2001.
25. Faiz Ö, Colak A, Saglam N et al. Determination and characterization of thermostable esterolytic activity from a novel thermophilic bacterium Anoxybacillus gonensis A4. J Biochem Mol Biol 40: 588-594, 2007.
26. Ozel A, Colak A, Arslan O et al. Purifi cation and characterisation of a polyphenol oxidase from Boletus erythropus and investigation of its catalytic effi ciency in selected organic solvents. Food Chem 119: 1044-1049, 2010.
27. Colak, A, Kolcuoglu Y, Faiz Ö et al. Comparative characterization of diphenolases from two mulberry fruits (Morus alba L. and Morus nigra L.). Asian J Chem 19: 2961- 2972, 2007.
28. Colak A, Şişik D, Saglam N et al. Characterization of a thermoalkalophilic esterase from a novel thermophilic bacterium, Anoxybacillus gonensis G2. Bioresour Technol 96: 625-631, 2005.
29. Ozen A, Colak A, Dincer B et al. A diphenolase from persimmon fruits (Diospyros kaki L., Ebenaceae). Food Chem 85: 431-437, 2004.
30. Ohnishi K, Yohko Y, Sekiguchi J. Purifi cation and characterization of a novel lipolytic enzyme from Aspergillus oryzae. J Ferment Bioeng 77: 490-495, 1994.
31. Dalmau E, Montesinos JL, Lotti M et al. Eff ect of diff erent carbon sources on lipase production by Candida rugosa. Enzyme Microbiol Technol 26: 657-63, 2000.
32. Ift ikhar T, Hussain A. Eff ects of nutrients on the extracellular lipase production by mutant strain of Rhizopus oligosporus TUV-31. Biotechnology 1: 15-20, 2002.
33. Chander H, Batish VK, Sannabhadti SS et al. Factors aff ecting lipase production in Aspergillus wentii. J Food Sci 45: 598-600, 1980.
34. Akhtar MW, Mirza AQ, Chughtai MID. Lipase induction in Mucor hiemalis. Appl Environ Microbiol 44: 257-63, 1980.
35. Rathi P, Saxena RK, Gupta R. A novel alkaline lipase from Burkholderia cepacia for detergent formulation. Process Biochem 37: 187-92, 2001.
36. Zhang HY, Wang X, Ching CB et al. Experimental optimization of enzymic kinetic resolution of racemic fl urbiprofen. Biotechnol Appl Bioc 42: 67-71, 2005.
37. Hiol A, Jonzo MD, Druet D et al. Production, purifi cation and characterization of an extracellular lipase from Mucor hiemalis f. hiemalis. Enzyme Microb Tech 25: 80-87, 1999.
38. Saxena RK, Davidson WS, Sheoran A et al. Purifi cation and characterization of an alkaline thermostable lipase from Aspergillus carneus. Process Biochem 39: 239-247, 2003.
39. Alhir S, Markakis P, Chadan RC. Lipase of Penicillium caseicolum. J Agric Food Chem 38: 598-601, 1990.
40. Savitha J, Srividya S, Jagat R et al. Identifi cation of potential fungal strain(s) for the production of inducible, extracellular and alkalophilic lipase. Afr J Biotechnol 6: 564-568, 2007.
41. Brush TS, Chapman R, Kurzman R et al. Purifi cation and characterization of extracellular lipases from Ophiostoma piliferum. Bioorg Med Chem 7: 2131-2138, 1999.
42. Hiol A, Jonzo MD, Rugani N et al. Purifi cation and characterization of an extracellular lipase from a thermophilic Rhizopus oryzae strain isolated from palm fruit. Enzyme Microb Tech 26: 421-430, 2000.
43. Costa MAF, Peralta RM. Production of lipase by soil fungi and partial characterization of lipase from a selected strain (Penicillium wortmanii). J Basic Microb 39: 11-16, 1999.
44. Abbas H, Hiol A, Deyris V et al. Isolation and characterization of an extracellular lipase from Mucor sp. strain isolated from palm fruit. Enzyme Microb Tech 31: 968-975, 2002.
45. Kashmiri MA, Adnan A, Butt BW. Production, purifi cation and partial characterization of lipase from Trichoderma viride. Afr J Biotechnol 5: 878-882, 2006.
46. Hasan F, Shah AA, Hameed A. Industrial applications of microbial lipases. Enzyme Microb Tech 39: 235-251, 2006.
47. Suzuki M, Yamamoto H, Mizugaki M. Purifi cation and general properties of a metal-insensitive lipase from Rhizopus japonicus NR 400. J Biochem 100: 1207-1213, 1986.
48. Yu XW, Wang LL, Yan X. Rhizopus chinensis lipase: Gene cloning, expression in Pichia pastoris and properties. J Mol Catal B-Enzym 57: 304-311, 2009.
49. Bancerz R, Ginalska G. A novel thermostable lipase from Basidiomycete Bjerkandera adusta R59: Characterization and esterifi cation studies. J Ind Microbiol Biot 34: 553-560, 2007.

APA	ÜLKER S, Özel A, Çolak A, Alpay Karaoğlu Ş (2011). Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. , 543 - 550.
Chicago	ÜLKER Serdar,Özel Arzu,Çolak Ahmet,Alpay Karaoğlu Şengül Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. (2011): 543 - 550.
MLA	ÜLKER Serdar,Özel Arzu,Çolak Ahmet,Alpay Karaoğlu Şengül Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. , 2011, ss.543 - 550.
AMA	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. . 2011; 543 - 550.
Vancouver	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. . 2011; 543 - 550.
IEEE	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş "Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil." , ss.543 - 550, 2011.
ISNAD	ÜLKER, Serdar vd. "Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil". (2011), 543-550.

APA	ÜLKER S, Özel A, Çolak A, Alpay Karaoğlu Ş (2011). Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. Turkish Journal of Biology, 35(5), 543 - 550.
Chicago	ÜLKER Serdar,Özel Arzu,Çolak Ahmet,Alpay Karaoğlu Şengül Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. Turkish Journal of Biology 35, no.5 (2011): 543 - 550.
MLA	ÜLKER Serdar,Özel Arzu,Çolak Ahmet,Alpay Karaoğlu Şengül Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. Turkish Journal of Biology, vol.35, no.5, 2011, ss.543 - 550.
AMA	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. Turkish Journal of Biology. 2011; 35(5): 543 - 550.
Vancouver	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil. Turkish Journal of Biology. 2011; 35(5): 543 - 550.
IEEE	ÜLKER S,Özel A,Çolak A,Alpay Karaoğlu Ş "Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil." Turkish Journal of Biology, 35, ss.543 - 550, 2011.
ISNAD	ÜLKER, Serdar vd. "Isolation, production, and characterization of an extracellular lipase from Trichoderma harzianum isolated from soil". Turkish Journal of Biology 35/5 (2011), 543-550.