Suraini Abd-Aziz
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Teoh Lay Sin
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Noorjahan Alitheen
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Neelam Shahab
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Kamarulzaman Kamaruddin
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
ABSTRACT
The current increase in amount of seafood wastes produced by the shrimp industry has lead to the finding of new methods for shrimp waste disposal or waste reused. For this respect, chitinase-producing fungi have been extensively studied as biocontrol agents. Locally isolated Trichoderma viren UKM1 was used in this study. From preliminary study, commercialized Trichoderma Minimal Medium (TMM) was selected for the degradation study. The substrates used were colloidal chitin as control substrate, sun dried ground and unground shrimp shells. Scanning Electron Microscopy (SEM) studies showed penetration of fungus mycelium into the colloidal chitin as compare to sun dried ground and unground. This observation suggested that colloidal chitin was the best carbon source for modeling the degradation of chitin materials. Stereo microscope studies suggested that the fungus removed (degraded) the chitinous materials layer by layer as indicated by the significant reduction in shell thickness. Shrimp shells were further evaluated for end products in the crude medium using High Performance Liquid Chromatography (HPLC). A simple, rapid, selective and specific HPLC method was developed to quantify glucosamine indirectly using the value of total N-acetyl-glucosamine (NAG) produced which the production of chitooligomer was used as marker. Results showed that the Trichoderma virens UKM1 secretes a significant amount of exochitinase compared to endochitinase by the identification of monomeric N-acetyl-glucosamine (NAG) from the chitinous substrate. The highest specific enzyme activity obtained using colloidal chitin was 14.59 U mg-1. Percentage of residual chitooligomer in impure chitinases samples was 86%.
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How to cite this article
Suraini Abd-Aziz, Teoh Lay Sin, Noorjahan Alitheen, Neelam Shahab and Kamarulzaman Kamaruddin, 2008. Microbial Degradation of Chitin Materials by Trichoderma virens UKM1. Journal of Biological Sciences, 8: 52-59.
DOI: 10.3923/jbs.2008.52.59
URL: https://scialert.net/abstract/?doi=jbs.2008.52.59
DOI: 10.3923/jbs.2008.52.59
URL: https://scialert.net/abstract/?doi=jbs.2008.52.59
REFERENCES
- Andronopoulou, E. and C.E. Vorgias, 2004. Multiple components and induction mechanism of the chitinolytic system of the hyperthermophilic archaeon Thermococcus chitonophagus. Applied Microb. Biotechnol., 65: 694-702.
Direct Link - Bolar, J.P., J.L. Norelli, G.E. Harman, S.K. Brown and H.S. Aldwinckle, 2001. Synergistic activity of endochitinase and exochitinase from Trichoderma atroviride (T. harzianum) against the pathogenic fungus (Venturia inaequalis) in transgenic apple plants. Trans. Res., 10: 533-543.
Direct Link - Dahiya, N., R. Tewari, R.P. Tiwari and G.S. Hoondal, 2005. Chtinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern. Electronic J. Biotechnol., 8: 134-145.
Direct Link - Dahiya, N., R. Tewari and G.S. Hoondal, 2005. Biotechnological aspects of chitinolytic enzymes: A review. Applied Microb. Biotechnol., 71: 773-782.
Direct Link - Deane, E.E., J.M. Whipps, J.M. Lynch and J.F. Peberdy, 1998. The purification and characterization of a Trichoderma harzianum exochitinase. Biochim. Biophys., Act., 1383: 101-110.
CrossRefDirect Link - Donzelli, B.G.G., G. Ostroff and G.E. Harman, 2003. Enhanced enzymatic hydrolysis of langostino shell chitin with mixtures of enzymes from bacterial and fungal sources. Carbohydr. Res., 338: 1833-1923.
CrossRefPubMedDirect Link - Ekblad, A. and T. Näsholm, 1996. Determination of chitin in fungi and mycorrhizal roots by an improved HPLC analysis of glucosamine. Plant Soil., 178: 29-35.
CrossRefDirect Link - Felse, P.A. and T. Panda, 2000. Production of microbial chitinases: A revisit. Bioprocess Eng., 23: 127-134.
Direct Link - Gómez-Ramírez, M., L.I. Rojas-Avelizapa, N.G. Rojas-Avelizapa and R. Cruz-Camarillo, 2004. Colloidal chitin stained with Remazol Brilliant Blue R®, a useful substrate to select chitinolytic microorganisms and to evaluate chitinases. J. Microbiol. Methods, 56: 213-219.
Direct Link - Havukkala, I., C. Mitamura, S. Hara, K. Hirayae, Y. Nishizawa and T. Hibi, 1993. Induction and purification of Beauveria bassiana chitinolytic enzymes. J. Invertbr. Pathol., 61: 97-102.
CrossRefDirect Link - Horn, S.J., A. Sorbotten, B. Synstad, P. Sikorski, M. Sorlie, K.M. Varum and V.G. Eijsink, 2006. Endo/exo mechanism and processivity of family 18 chitinases produced by Serratia marcescens. FEBS J., 273: 491-503.
Direct Link - Kumar, M.N.V.R., 2000. A review of chitin and chitosan applications. React. Funct. Polym., 46: 1-27.
CrossRefDirect Link - Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall, 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275.
CrossRefPubMedDirect Link - Madhavan, N.K. and P. Ashok, 1996. Solid state fermentation for L-glutamic acid production using Brevibacterium sp. Biotechnol. Lett., 18: 199-204.
Direct Link - Mislovicova, D., J. Masarova, K. Bendzalova, L. Soltes and E. Machova, 2000. Sonication of chitin-glucan, preparation of water-soluble fractions and characterization by HPLC. Ultrason. Sonochem., 7: 63-68.
Direct Link - Park, J.K., K. Morita, I. Fukumoto, Y. Yamasaki, T. Nakagawa, M. Kawamukai and H. Matsuda, 1997. Purification and characterization of the Chitinase (ChiA) from Enterobacter sp. G-l. Biosci. Biotechnol. Biochem., 61: 684-689.
CrossRefDirect Link - Punin-Crespo, M.O., M. Vilasoa-Martinez, J. Lopez-Hernandez and M.A. Lage-Yusty, 2006. High-performance liquid chromatographic determination of chitin in the snow crab, Chionoecetes opilio. J. Chromatogr., 1116: 189-192.
Direct Link - Ren, Y., K.E. Wee and F.N. Chang, 2000. Deficiency of current methods in assaying endochitinase activity. Biochem. Biophys. Res. Commun., 268: 302-305.
Direct Link - Studer, M., K. Fluck and W. Zimmermann, 1992. Production of chitinases by Aphanocladium album grown on crystalline and colloidal chitin. FEMS Microbiol. Lett., 99: 213-216.
Direct Link - Suginta, W., A. Vongsuwan, C. Songsiriritthigul and J. Svasti, 2005. Enzymatic properties of wild-type and active site mutants of chitinase A from Vibrio carchariae, as revealed by HPLC-MS. FEBS J., 272: 3376-3386.
Direct Link - Takayanagi, T., K. Ajisaka, Y. Takiguchi and K. Shimahara, 1991. Isolation and characterization of thermostable chitinases from Bacillus licheniformis X-7u. Biochem. Biophys. Acta (BBA)-Protein Struct. Mol. Enzymol., 1078: 404-410.
CrossRefDirect Link - Wang, S.L. and J.R. Hwang, 2000. Microbial reclamation of shellfish wastes for the production of chitinases. Enzyme. Microb. Technol., 28: 376-382.
Direct Link - Wang, S.Y., A.L. Moyne, G. Thottappilly, S.J. Wu, R.D. Locy and N.K. Singh, 2001. Purification and characterisation of a Bacillus cereus exochitinase. Enzyme. Microbial. Technol., 28: 492-498.
Direct Link - Wang, S.L., T.Y. Lin, Y.H. Yen, H.F. Liao and Y.J. Chen, 2006. Bioconversion of shellfish chitin wastes for the production of Bacillus subtilis W-118 chitinase. Carbohydrate Res., 341: 2507-2515.
CrossRef - Zhang, J. and R. Greasham, 1999. Chemically defined media for commercial fermentations. Applied Microbiol. Biotechnol., 51: 407-421.
CrossRef - Zhu, X., J. Cai, J. Yang and Q. Su, 2005. Determination of glucoamine in impure chitin samples by high-performance liquid chromatography. Carbohydrate Res., 340: 1732-1738.
Direct Link