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Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​

Year 2016, Volume: 44 Issue: 1, 65 - 75, 01.03.2016

Abstract

In this study, pectin lyase PL was obtained from Aspergillus fumigatus strain 2101 which was isolated from local water sources. PL was purified using ammonium sulphate precipitation, gel filtration and ion-exchange chromatography. After these purification steps, 12.4 purification fold was obtained. The optimum pH and tem- perature of PL were 8.0 and 40 ̊C at 70 min, respectively. The molecular weight of PL was 11.5 kDa. Maximum PL activity was obtained with fruit pulps in submerged culture medium. Stability assays showed that the PL was stable till 20 months at 4 ̊C. The highest PL activity was obtained using citrus pectin as carbon source. PL was strongly activated by FeSO4, FeCl3 and ascorbic acid.

References

  • 1. J.N. Be Miller, An introduction to pectins: Structure and properties, In Fishman, M.L., & Jem, J (Eds.), Chemistry and Functions of Pectins, ACS Symposium Series 310. American Chemical Society, Washington, DC, (1986).
  • 2. L. Delgado, B.A. Trejo, C. Huitrón, G. Aguilar, Pectin lyase from Aspergillus sp. CH-Y-1043, Applied Microbiology Biotechnology, 39 (1992) 515-519.
  • 3. R.P. De Vries, and J. Visser, Aspergillus enzymes involved in degradation of plant cell wall polysaccharides, Microbiology and Molecular Biology Reviews, 65 (2001) 497-552.
  • 4. A. Martínez-Trujillo, J.S. Aranda, C. Gómez-Sánchez, B. Trejo-Aguilar, G. Aguilar-Osorio, Constitutive and inducible pectinolytic enzymes from Aspergillus flavipes FP-500 and their modulation by pH and carbon source, Brazilian Journal of Microbiology, 40 (2009) 40-47.
  • 5. D.R. Kashyap, P.K. Vohra, S. Chopra, R. Tewari, Applications of pectinases in the commercial sector: a review, Bioresource Technology, 77 (2001) 215-227.
  • 6. R.S.S. Jayani, S. Saxena, R. Gupta, Microbial pectinolytic enzymes: A review. Process Biochemistry, 40 (2005) 2931–2944.
  • 7. S.R. Nair, S.K. Rakshit, T. Panda, Effect of carbon sources on the synthesis of pectinase by Aspergilli, Bioprocess Engineering, 13 (1995) 37-40.
  • 8. I. Reid, and M. Ricard, Pectinase in papermaking:solving retention problems in mechanical pulps bleached with hydrogen peroxide, Enzyme and Microbial Technology, 26 (2000) 115–123.
  • 9. S. Yadav, P.K. Yadav, D. Yadav, K.D.S. Yadav, Pectin lyase: A review, Process Biochemistry, 44 (2009) 1–10.
  • 10. I.G. Sandri, R.C. Fontana, D.M. Barfknecht, M.M. Silveira, Clarification of fruit juices by fungal pectinases, LWT-Food Science and Technology, 44 (2011) 2217–2222.
  • 11. P. Albersheim, Pectin lyase from fungi, Methods in Enzymology, 8 (1966) 628–631.
  • 12. U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of Bacteriophage T4, Nature, 15 (1970) 680-685.
  • 13. G. Temizkan, S. Yılmazer, M. Ozturk, S. Arı, H. Ertan, A. Topal-Sarıkaya, N. Arda, Moleküler Biyolojide Kullanılan Yöntemler. In: G Temizkan & N Arda (Eds.), Proteinlerin İzolasyonu, Analizi, Saflaştırılması, (2008) (pp. 161-273). İstanbul, Turkey.
  • 14. M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72 (1976) 248-254.
  • 15. D. Silva, E.S. Martins, R. Silva, E. Gomes, Pectinase production by Penicillium viridicatum RFC3 by solid state fermentation using agricultural wastes and agro-industrial by-products, Brazilian Journal of Microbiology, 33 (2002) 318-324.
  • 16. N. Martin, S.R. Souza, R. Silva, E. Gomes, Pectinase production by fungal strains in solid-state fermentation using agro-industrial bioproduct, Brazilian Archives of Biology and Technolgy, 47 (2004) 813-819.
  • 17. A. Alaña, A. Gabilondo, F. Hernando, M.D. Moragues, J.B. Domínguez, M.J. llama, J.L. Serra, Pectin Lyase Production by a Penicillium italicum strain, Applied and Environmental Microbiology, 55 (1989) 1612-1616.
  • 18. S. Koser, Z. Anwar, Z. Iqbal, A. Anjumc, T. Aqil, S. Mehmood, M. Irshada, Utilization of Aspergillus oryzae to produce pectin lyase from various agroindustrial residues, J of Radiation Research and Applied Sciences, 7 (2014) 327-332.
  • 19. S. Solís, J. Loeza, G. Segura, J. Tello, N. Reyes, P. Lappe, L. Guitérrez, F. Ríos, C. Huitrón, Hydrolysis of orange peel by a pectin lyase-overproducing hybrid obtained by protoplast fusion between mutant pectinolytic Aspergillus flavipes and Aspergillus niveus CH-Y-1043, Enzyme and Microbial Techology, 44 (2009) 123–128.
  • 20. C.C.H. Sakiyama, E.M. Paula, P.C. Pereira, A.C. Borges, D.O. Silva, Characterization of pectin lyase produced by an endophytic strain isolated from coffee cherries, Letters in Applied Microbiology, 33 (2001) 117-121.
  • 21. A. Alaña, I. Alkorta, J.B. Domínguez, M.J. Llama, J.L. Serra, Pectin lyase activity in a Penicillium italicum strain, Applied Environmental Microbiology, 56 (1990) 3755-3759.
  • 22. D.R. Kashyap, S. Chandra, A. Kaul, R. Tewari, Production, purification and characterization of pectinase from a Bacillus sp. DT7, World Journal of Microbiology and Biotechnology, 16 (2000) 277-282.
  • 23. S. Yadav, P.K. Yadav, D. Yadav, K.DS. Yadav, Purification and characterization of an alkaline pectin lyase from Aspergillus flavus, Process Biochemistry, 43 (2008) 547–552.
  • 24. S.N. Gummadi, and D.S. Kumar, Optimization of chemical and physical parameters affecting the activity of pectin lyase and pectatelyase from Debaryomycesflavusnepalensis: A statistical approach, Biochemical Engineering Journal, 30 (2006) 130–137.
  • 25. Q.K. Beg, B. Bhushan, M. Kapoor, G.S. Hoondal, Production and characterization of thermostable xylanase and pectinase from Streptomyces sp. QG-11- 3, Journal of Industrial Microbiology & Biotechnology, 24 (2000) 396-402.
  • 26. A. Alaña, M.J. Llama, J.L. Serra, Purification and some properties of the pectin lyase from Penicillium italicum, FEBS Letters, 280 (1991) 335-340.
  • 27. C. Perez-Fuentes, M.C. Ravanal, J.H. Eyzaguirre, Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization, Fungal Biology, 118 (2014) 507-515.
  • 28. S.H. Hamdy, Purification and characterization of the pectin lyase produced by Rhizopus oryzae grown on orange peels, Annals of Microbiology, 55 (2005). 205-211.
  • 29. S. Yadav, A.K. Dubey, G. Anand, R. Kumar, D. Yadav, Purification and biochemical characterization of an alkaline pectin lyase from Fusarium decemcellulare MTCC 2079 suitable for Crotolaria juncea fiber retting, Journal of Basic Microbiology, 53 (2013) 1-9.
  • 30. A. Losonczi, E. Csisza´r, G. Szaka´cs, L. Bezur, Role of the EDTA chelating agent in bioscouring of cotton, Textile Research Journal, 75 (2005). 411–417.
  • 31. W.C. Chen, E.C. Hsieh, T.C. Tseng, Purification and characterization of a pectin lyase from Pythium splendens infected cucumber fruits, Botanical Bulletin Academia Sinica, 39 (1998) 81-186.

Yerel Su İzolatı Aspergillus Fumigatus 2101 Suşu Tarafından Atık Malzemeler ile Pektinaz Üretimi ve Enzimin Saflaştırılması ve Karakterizasyonu

Year 2016, Volume: 44 Issue: 1, 65 - 75, 01.03.2016

Abstract

B u çalışmada, pektin liyaz PL yerel su kaynaklarından izole edilen Aspergillus fumigatus 2101 suşundan elde edildi. PL, amonyum sülfat çöktürmesi, jel filtrasyon ve iyon değişim kromatografisi kullanılarak saflaştırıldı. Bu saflaştırma basamaklarından sonra, 12.4 saflaştırma katsayısı elde edildi. PL’nin optimum pH ve sıcaklığı sırasıyla 8.0 ve 40˚C’de 70 dakikadır. PL’nin molekül ağırlığı 11.5 kDa’dur. Sıvı besi ortamında maksimum PL aktivitesi meyve pulpları ile elde edilmiştir. Stabilite deneyleri, PL’nin 4˚C’de 20 aya kadar kararlı olduğunu göstermiştir. En yüksek PL aktivitesi karbon kaynağı olarak sitrus pectin kullanıldığında elde edilmiştir. PL, FeSO, FeCl ve askorbik asit tarafından güçlü bir şekilde aktive edilmiştir

References

  • 1. J.N. Be Miller, An introduction to pectins: Structure and properties, In Fishman, M.L., & Jem, J (Eds.), Chemistry and Functions of Pectins, ACS Symposium Series 310. American Chemical Society, Washington, DC, (1986).
  • 2. L. Delgado, B.A. Trejo, C. Huitrón, G. Aguilar, Pectin lyase from Aspergillus sp. CH-Y-1043, Applied Microbiology Biotechnology, 39 (1992) 515-519.
  • 3. R.P. De Vries, and J. Visser, Aspergillus enzymes involved in degradation of plant cell wall polysaccharides, Microbiology and Molecular Biology Reviews, 65 (2001) 497-552.
  • 4. A. Martínez-Trujillo, J.S. Aranda, C. Gómez-Sánchez, B. Trejo-Aguilar, G. Aguilar-Osorio, Constitutive and inducible pectinolytic enzymes from Aspergillus flavipes FP-500 and their modulation by pH and carbon source, Brazilian Journal of Microbiology, 40 (2009) 40-47.
  • 5. D.R. Kashyap, P.K. Vohra, S. Chopra, R. Tewari, Applications of pectinases in the commercial sector: a review, Bioresource Technology, 77 (2001) 215-227.
  • 6. R.S.S. Jayani, S. Saxena, R. Gupta, Microbial pectinolytic enzymes: A review. Process Biochemistry, 40 (2005) 2931–2944.
  • 7. S.R. Nair, S.K. Rakshit, T. Panda, Effect of carbon sources on the synthesis of pectinase by Aspergilli, Bioprocess Engineering, 13 (1995) 37-40.
  • 8. I. Reid, and M. Ricard, Pectinase in papermaking:solving retention problems in mechanical pulps bleached with hydrogen peroxide, Enzyme and Microbial Technology, 26 (2000) 115–123.
  • 9. S. Yadav, P.K. Yadav, D. Yadav, K.D.S. Yadav, Pectin lyase: A review, Process Biochemistry, 44 (2009) 1–10.
  • 10. I.G. Sandri, R.C. Fontana, D.M. Barfknecht, M.M. Silveira, Clarification of fruit juices by fungal pectinases, LWT-Food Science and Technology, 44 (2011) 2217–2222.
  • 11. P. Albersheim, Pectin lyase from fungi, Methods in Enzymology, 8 (1966) 628–631.
  • 12. U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of Bacteriophage T4, Nature, 15 (1970) 680-685.
  • 13. G. Temizkan, S. Yılmazer, M. Ozturk, S. Arı, H. Ertan, A. Topal-Sarıkaya, N. Arda, Moleküler Biyolojide Kullanılan Yöntemler. In: G Temizkan & N Arda (Eds.), Proteinlerin İzolasyonu, Analizi, Saflaştırılması, (2008) (pp. 161-273). İstanbul, Turkey.
  • 14. M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72 (1976) 248-254.
  • 15. D. Silva, E.S. Martins, R. Silva, E. Gomes, Pectinase production by Penicillium viridicatum RFC3 by solid state fermentation using agricultural wastes and agro-industrial by-products, Brazilian Journal of Microbiology, 33 (2002) 318-324.
  • 16. N. Martin, S.R. Souza, R. Silva, E. Gomes, Pectinase production by fungal strains in solid-state fermentation using agro-industrial bioproduct, Brazilian Archives of Biology and Technolgy, 47 (2004) 813-819.
  • 17. A. Alaña, A. Gabilondo, F. Hernando, M.D. Moragues, J.B. Domínguez, M.J. llama, J.L. Serra, Pectin Lyase Production by a Penicillium italicum strain, Applied and Environmental Microbiology, 55 (1989) 1612-1616.
  • 18. S. Koser, Z. Anwar, Z. Iqbal, A. Anjumc, T. Aqil, S. Mehmood, M. Irshada, Utilization of Aspergillus oryzae to produce pectin lyase from various agroindustrial residues, J of Radiation Research and Applied Sciences, 7 (2014) 327-332.
  • 19. S. Solís, J. Loeza, G. Segura, J. Tello, N. Reyes, P. Lappe, L. Guitérrez, F. Ríos, C. Huitrón, Hydrolysis of orange peel by a pectin lyase-overproducing hybrid obtained by protoplast fusion between mutant pectinolytic Aspergillus flavipes and Aspergillus niveus CH-Y-1043, Enzyme and Microbial Techology, 44 (2009) 123–128.
  • 20. C.C.H. Sakiyama, E.M. Paula, P.C. Pereira, A.C. Borges, D.O. Silva, Characterization of pectin lyase produced by an endophytic strain isolated from coffee cherries, Letters in Applied Microbiology, 33 (2001) 117-121.
  • 21. A. Alaña, I. Alkorta, J.B. Domínguez, M.J. Llama, J.L. Serra, Pectin lyase activity in a Penicillium italicum strain, Applied Environmental Microbiology, 56 (1990) 3755-3759.
  • 22. D.R. Kashyap, S. Chandra, A. Kaul, R. Tewari, Production, purification and characterization of pectinase from a Bacillus sp. DT7, World Journal of Microbiology and Biotechnology, 16 (2000) 277-282.
  • 23. S. Yadav, P.K. Yadav, D. Yadav, K.DS. Yadav, Purification and characterization of an alkaline pectin lyase from Aspergillus flavus, Process Biochemistry, 43 (2008) 547–552.
  • 24. S.N. Gummadi, and D.S. Kumar, Optimization of chemical and physical parameters affecting the activity of pectin lyase and pectatelyase from Debaryomycesflavusnepalensis: A statistical approach, Biochemical Engineering Journal, 30 (2006) 130–137.
  • 25. Q.K. Beg, B. Bhushan, M. Kapoor, G.S. Hoondal, Production and characterization of thermostable xylanase and pectinase from Streptomyces sp. QG-11- 3, Journal of Industrial Microbiology & Biotechnology, 24 (2000) 396-402.
  • 26. A. Alaña, M.J. Llama, J.L. Serra, Purification and some properties of the pectin lyase from Penicillium italicum, FEBS Letters, 280 (1991) 335-340.
  • 27. C. Perez-Fuentes, M.C. Ravanal, J.H. Eyzaguirre, Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization, Fungal Biology, 118 (2014) 507-515.
  • 28. S.H. Hamdy, Purification and characterization of the pectin lyase produced by Rhizopus oryzae grown on orange peels, Annals of Microbiology, 55 (2005). 205-211.
  • 29. S. Yadav, A.K. Dubey, G. Anand, R. Kumar, D. Yadav, Purification and biochemical characterization of an alkaline pectin lyase from Fusarium decemcellulare MTCC 2079 suitable for Crotolaria juncea fiber retting, Journal of Basic Microbiology, 53 (2013) 1-9.
  • 30. A. Losonczi, E. Csisza´r, G. Szaka´cs, L. Bezur, Role of the EDTA chelating agent in bioscouring of cotton, Textile Research Journal, 75 (2005). 411–417.
  • 31. W.C. Chen, E.C. Hsieh, T.C. Tseng, Purification and characterization of a pectin lyase from Pythium splendens infected cucumber fruits, Botanical Bulletin Academia Sinica, 39 (1998) 81-186.
There are 31 citations in total.

Details

Primary Language English
Journal Section Research Article
Authors

Hacer Yamancı This is me

Kader Poturcu This is me

Ismail Ozmen This is me

Hacı Halil Bıyık This is me

Publication Date March 1, 2016
Published in Issue Year 2016 Volume: 44 Issue: 1

Cite

APA Yamancı, H., Poturcu, K., Ozmen, I., Bıyık, H. H. (2016). Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​. Hacettepe Journal of Biology and Chemistry, 44(1), 65-75.
AMA Yamancı H, Poturcu K, Ozmen I, Bıyık HH. Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​. HJBC. March 2016;44(1):65-75.
Chicago Yamancı, Hacer, Kader Poturcu, Ismail Ozmen, and Hacı Halil Bıyık. “Pectinase Production With Waste Materials by Local Water Isolate of Aspergillus Fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​”. Hacettepe Journal of Biology and Chemistry 44, no. 1 (March 2016): 65-75.
EndNote Yamancı H, Poturcu K, Ozmen I, Bıyık HH (March 1, 2016) Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​. Hacettepe Journal of Biology and Chemistry 44 1 65–75.
IEEE H. Yamancı, K. Poturcu, I. Ozmen, and H. H. Bıyık, “Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​”, HJBC, vol. 44, no. 1, pp. 65–75, 2016.
ISNAD Yamancı, Hacer et al. “Pectinase Production With Waste Materials by Local Water Isolate of Aspergillus Fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​”. Hacettepe Journal of Biology and Chemistry 44/1 (March 2016), 65-75.
JAMA Yamancı H, Poturcu K, Ozmen I, Bıyık HH. Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​. HJBC. 2016;44:65–75.
MLA Yamancı, Hacer et al. “Pectinase Production With Waste Materials by Local Water Isolate of Aspergillus Fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​”. Hacettepe Journal of Biology and Chemistry, vol. 44, no. 1, 2016, pp. 65-75.
Vancouver Yamancı H, Poturcu K, Ozmen I, Bıyık HH. Pectinase Production with Waste Materials by Local Water Isolate of Aspergillus fumigatus Strain 2101 and Purification and Characterization of the Enzyme ​. HJBC. 2016;44(1):65-7.

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