Uğur ÇÖMLEKÇİOĞLU, FERİT CAN YAZDIÇ, Sevtap KESER, Bekir Mehmet KELLECİ, Gülay BATTALOĞLU, Emin ÖZKÖSE

Neocallimastix sp. ve orpinomyces sp.’nin enzim üretimleri üzerine karbon kaynaklarının etkisi

Neocallimastix sp. ve polisentrik Orpinomyces sp. 12 ticari karbon kaynağı ve 11 bitkisel karbon kaynağı üzerinde geliştirilerek β-1,4-endoglukanaz, β-1,4-endoksilanaz, β-glukosidaz, β-ksilosidaz üretimleri belirlenmiştir. Enzimler üçüncü günden itibaren besi ortamında toplanmış, en yüksek selülaz aktivite değerleri Neocallimastix sp. ve Orpinomyces sp.’de sırasıyla karboksimetil selüloz ve tekstil atığından elde edilmiştir. Kolay fermente edilebilen şekerler selüloz üretimini 0.5 U/ml altında kalmasına neden olmuştur. Ksilanaz en yüksek Neocallimastix sp.’nin karboksimetil selüloz kültüründe, Orpinomyces sp.’nin fruktoz, maltoz, sükroz, laktoz ve avisel kültürlerinde tespit edilmiş, ayrıca yonca her iki fungus içinde ksilanaz aktivitesini en üst düzeye çıkarmıştır. Neocallimastix sp.’de b-glikosidaz ve b-ksilosidaz üretimi Orpinomyces sp.’ye göre daha fazla olmuştur. İnulin her iki fungus tarafından fermente edilse de fungal gelişimleri dolayısıyla da enzim üretimlerini olumsuz etkileyen tek substrat olmuştur. Ksilanazın sürekli salgılanma doğası anaerobik fungusların konak hayvanın hemiselülozdan faydalanmasında önemli rol oynadığını göstermektedir.

Anahtar Kelimeler:

inülin, beta-glukozidaz, Neocallimastix, kuru ot, yonca, yonca kuru otu, ksilan 1, 4-beta-ksilosidaz, karbon

Effects of carbon sources on enzyme production of neocallimastix sp. ve orpinomyces sp.

Neocallimastix sp. and Orpinomyces sp. was grown by using 12 commercial carbon sources and 11 plant carbon sources. β-1,4-endoglucanase, β-1,4-endoxylanase, β-glucosidase, β-xylosidase synthesis by these fungi were determined. Enzymes accumulated on culture medium after three days. The maximum cellulase activities of Neocallimastix sp. and Orpinomyces sp. was observed in the cultures of carboxymethyl cellulose and textile waste, respectively. In the cultures of readily fermentable sugars, cellulase activity was under 0.5 U/ml. Xylanase activity was found to be maximum in carboxymethyl cellulose culture of Neocallimastix sp. However maximum xylanase activities of Orpinomyces sp. was found in fructose, maltose, sucrose, lactose and avicel cultures. Xylanase production of both fungi was also induced by alfalfa hay. Neocallimastix sp. produced more β-glicosidase and β-xylosidase than Orpinomyces sp. Inulin was fermented by both fungi, however inulin was the only substrate that caused poor growth and enzyme production. The continuous secretion of xylanase reflected the role of anaerobic fungi in hemicellulose degradation in host animal.

Keywords:

inulin, beta-glucosidase, Neocallimastix, hay, lucerne, lucerne hay, xylan 1, 4-beta-xylosidase, carbon,

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1. Canbolat Ö: Potential nutritive value of field Binweed (Convolvulus arvensis L) Hay harvested at three different maturity stages. Kafkas Univ Vet Fak Derg, 18 (2): 331-335, 2012.
2. Thomson JA: Molecular biology of xylan degradation. FEMS Microbiol Rev, 104, 65-82, 1993.
3. Pearce PD, Bauchop T: Glycosidases of the rumen anaerobic fungus Neocallimastix frontalis grown on cellulosic substrates. Appl Environ Microbiol, 49, 1265-1269, 1985.
4. Lee SS, Shin KJ, Kim WY, Ha JK, Han IK: The rumen ecosystem: As a fountain source of nobel enzymes. Asian-Aus J Anim Sci, 12 (6): 988-1001, 1999.
5. Cheng KJ, Forsberg CW, Minato H, Costerton JW: Microbial ecology and physiology of feed degradation within the rumen. In, Tsuda T, Sasaki H, Kawahima R (Eds): Physiological Aspects of Digestion and Metabolism in Ruminants. pp. 595-624, Academic Press, New York, 1991.
6. Akin DE, Gordon GLR, Hogan JP: Rumen bacterial and fungal degradation of Digiteria pentzii grown with or without sulfur. Appl Environ Microbiol, 46, 738-748, 1983.
7. Wood TM, Wilson CA, Mccrea SI, Joblin KN: A highly active extracellular cellulase from the anaerobic rumen fungus Neocallimastix frontalis. FEMS Microbiol Lett, 34, 37-40, 1986.
8. Trinci APJ, Davis DR, Gull K, Lawrence MI, Nielsen BB, Rickers A, Theodorou MK: Anaerobic fungi in herbivorous animals. Mycol Res, 98, 129-152, 1994.
9. Williams AG, Orpin CG: Glycoside hydrolase enzymes present in the zoospore and vegetative growth stages of the rumen fungi Neocallimastix frontalis, Piromonas communis and an unidentified isolate grown on a range of carbohydrate substrates, Can J Microbiol, 33 (5): 427-434, 1987.
10. Çömlekçioğlu U, Akyol İ, Kar B, Özköse E ve Ekinci MS: Anaerobik rumen funguslarının izolasyonu, tanımlanması ve kültür koleksiyonunun oluşturulması. Hayvansal Üretim, 49 (2): 29-35, 2008.
11. Orpin CG: Studies on the rumen flagellate Sphaeromonas communis. J Gen Microbiol, 94, 270-280, 1976.
12. Miller GL: Use of dinitrosalicylic acid regent for determination of reducing sugars. Anal Chem, 31, 426-428, 1959.
13. Comlekcioglu U, Ozkose E, Yazdic FC, Akyol I, Ekinci MS: Poly-saccharidase and glycosidase production of avicel grown rumen fungus Orpinomyces sp. GMLF5. Acta Biol Hung, 61 (3): 333-343, 2010.
14. Borneman WS, Akin DE, Ljungdahl LG: Fermentation products and plant cell wall degrading enzymes produced by monocentric and polycentric ruminal fungi. Appl Environ Microbiol, 9, 285-296, 1989.
15. Comlekcioglu U, Aygan A, Yazdic FC, Ozkose E: Effects of various agro-wastes on xylanase and b-xylosidase production of anaerobic ruminal fungi. J Sci Ind Res, 70, 293-299, 2011.
16. Yanke LJ, Selinger LB, Lynn JR, Cheng KJ: Comparison of the influence of carbon substrates on the fibrolytic activities of anaerobic rumen fungi. Anaerobe, 2, 373-378, 1996.
17. Williams AG, Orpin CG: Polysaccharide-degrading enzymes formed by three species of anaerobic rumen fungi on a range of carbonhydrate substrates. Can J Microbiol, 33, 418-426, 1987.
18. Teunissen MJ, Hermans JMH, Huis In’t Veld JHS, Vogels GD: Purification and characterisation of a complex-bound and a free b-1,4-endoxylanase from the culture fluid of the anaerobic fungus Piromyces sp. Strain E2. Arch Microbiol, 159, 265-271, 1993.
19. Lowe SE, Theodorou MK, Trinci APJ: Cellulases and xylanases of an anaerobic rumen fungus grown on wheat straw, wheat straw holocellulose, cellulose and xylan. Appl Environ Microbiol, 53, 1216-1223, 1987.
20. Williams AG, Withers SE, Orpin CG: Effect of the carbohydrate growth substrate on polysaccharolytic enzyme formation by anaerobic fungi isolated from the foregut and hindgut of nonruminant herbivores and the forestomach of ruminants, Lett Appl Microbiol, 18, 147-151, 1994.
21. Mountfort DO, Asher RA: Production of xylanase by the ruminal anaerobic fungus Neocallimastix frontalis. Appl Environ Microbiol, 55, 1016-1022, 1989.
22. Phillips MW, Gordon GLR: Sugar and polysaccharide fermentation by anaerobic fungi from Australia, Britain and New Zealand. BioSystems 21, 377-383, 1988.
23. Paul SS, Kamra DN, Sastry VRB: Fermentative characteristics and fibrolytic activities of anaerobic gut fungi isolated from wild and domestic ruminants, Arch Anim Nutr, 64, 279-292, 2010.
24. Li X, Calza RE: Purification and characterization of an extracellular b-glucosidase from the rumen fungus Neocallimastix frontalis EB188. Enz Microbial Technol, 13, 1-7, 1991.
25. Chen H, Xinliang L, Ljungdahl LG: Isolation and Properties of an Extracellular b-glucosidase from the polycentric rumen fungus Orpinomyces sp. Strain PC-2. Appl Environ Microbiol, 60 (1): 64-70, 1994.