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Structural and Molecular Characterization of Extracellular Polysaccharides Produced by a New Fungal Strain, Trichoderma erinaceum DG-312  

JOO JI-HOON (Department of Biotechnology, Daegu University)
YUN JONG-WON (Department of Biotechnology, Daegu University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.6, 2005 , pp. 1250-1257 More about this Journal
Abstract
Two groups of exopolysaccharides (designated as Fr-I EPS and Fr-II EPS) were isolated from the culture filtrate of new fungal strain Trichoderma erinaceum DG-312 by Sepharose CL-6B chromatography. The structures of the exopolysaccharides were investigated using gas chromatography (GC), Fourier transform-infrared (FT-IR) spectroscopy, GCMS analysis, and NMR. GC analysis indicated that Fr-I EPS was composed of mainly mannose ($78.9\%$) and galactose ($21.1\%$), whereas Fr-II EPS contained mannose ($68.4\%$), galactose ($26.2\%$), and glucose ($5.4\%$). In the anomeric region ($950-700cm_{-1}$) of the FT-IR spectrum, both EPSs exhibited obvious characteristic absorption of $810\;cm_{-1}$, indicating the existence of mannose. The spectra of $\alpha-and\;\beta$-configurations were assigned at 880 and $914\;cm_{-1}$, respectively. The results of GC-MS analyses confirmed that both EPSs were complex heteropolysaccharides with a ($1{\rightarrow}3$)-linked mannan backbone. The C-1 region that appeared in the $^{13}C-NMR$ spectra of these EPSs indicated a typical anomeric carbon signal. The Fr-I EPS showed two anomeric carbon signals at 102.6 and 99.6 ppm, whereas the Fr-II EPS displayed four anomeric carbon signals at 102.5, 99.6, 98.5, and 94.3 ppm. The molecular characteristics of the EPSs were further investigated using a size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system. The SEC/MALLS system revealed that the average molar masses of the EPSs were $6.592{\times}10^{4}$ (Fr-I EPS) and $1.920{\times}10^{4}$ (Fr-II EPS) g/mol, and the molecular conformation of both EPSs in aqueous solution was random coils.
Keywords
Exopolysaccharides; GC-MS; NMR; SEC/MALLS; structural analys; Trichoderma erinaceum;
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1 Ahrazem, O., A. Prieto, J. A. Leal, J. J. Barbero, and M. Bernabe. 2002. Fungal cell-wall galactomannans isolated from Geotrichum spp. and their teleomorphs, Dipodascus and Galactomyces. Carbohydr. Res. 337: 2347-2351   DOI   ScienceOn
2 Duncan, C. J. G., N. Pugh, D. S. Pasco, and S. A. Ross. 2002. Isolation of a galactomannan that enhances macrophage activation from the edible fungus Morchella esculenta. J. Agric. Food Chem. 50: 5683-5685   DOI   ScienceOn
3 Ganter, J. L. M. S. and F. Reicher. 1999. Water-soluble galactomannans from seeds of Mimosaceae spp. Bioresource Technol. 68: 55-62   DOI   ScienceOn
4 Joshi, H. and V. P. Kapoor. 2003. Cassia grandis Linn. f. seed galactomannan: Structural and crystallographical studies. Carbohydr. Res. 338: 1907-1912   DOI   ScienceOn
5 Waytt, P. J. 1993. Light scattering and the absolute characterization of macromolecules. Anal. Chim. Acta 272: 1-40   DOI   ScienceOn
6 Zhang, M., L. Zhang, and P. C. K. Cheung. 2003. Molecular mass and chain conformation derivatives of $\beta$-glucan from sclerotia of Pleurotus tuber-regium. Biopolymers 68: 150-159   DOI   ScienceOn
7 Chaubey, M. and V. P. Kapoor. 2001. Structure of a galactomannan from the seeds of Cassia angustifolia Vahl. Carbohydr. Res. 332: 439-444   DOI   ScienceOn
8 Dubois, M., K. A. Gilles, J. K. Hamiton, P. A. Rebers, and F. Smith. 1956. Colorimetric method for determination of sugar and related substance. Anal. Chem. 28: 350-356   DOI
9 Reis, A. V., O. A. Cavalcanti, A. F. Rubrira, and E. C. Muniz. 2003. Synthesis and characterization of hydrogels formed from a glycidyl methacrylate derivative of galactomannan. Int. J. Pharmaceutics 267: 13-25   DOI   PUBMED   ScienceOn
10 Arja, M.-O., P. Marja, L. Raija, and S. Pirkko. 2005. Enhanced production of cellobiohydrolases in Trichoderma reesei and evaluation of the new preparations in biofinishing of cotton. J. Biotechnol. 116: 305-317   DOI   ScienceOn
11 Hakomori, S. I. 1964. A rapid permethylation of glycolipid and polysaccharide catalyzed by methysulfinyl carbanion in dimethyl sulfoxide. J. Biochem. 55: 205-208
12 Morris, E., O.P.E. Doyle, and K. J. Clancy. 1995. A profile of Trichoderma species. I. Mushroom compost production. Mushroom Sci. 14: 611-618
13 Eufrocinio, C. M., B. Takeshi, I. K. Shin, F. Ei-ichiro, and K. Akio. 2002. Biosynthetic studies of the tetramic acid antibiotic trichosetin. Tetrahedron 58: 6655-6658   DOI   ScienceOn
14 Grosev, V. M., R. Bozac, and G. J. Puppels. 2001. Vibrational spectroscopic characterization of wild growing mushroom and toadstools. Spectrochim. Acta Part A Mol. Biomol. Spectrosc. 57: 2815-2829   DOI   ScienceOn
15 Akerholm, M. and L. Salmen. 2001. Interactions between wood polymers studied by dynamic FT-IR spectroscopy. Polymer 42: 963-969   DOI   ScienceOn
16 Galichet, A., G. D. Sockalingum, A. Belarbi, and M. Manfait. 2001. FTIR spectroscopic analysis of Saccharomyces cerevisiae cell walls: Study of an anomalous strain exhibiting a pinkcolored cell phenotype. FEMS Microbiol. Lett. 197: 179-186   DOI   ScienceOn
17 Morris, E., O.P.E. Doyle, and K. J. Clancy. 1995. A profile of Trichoderma species. II. Mushroom growing units. Mushroom Sci. 14: 619-626
18 Zhang, P., L. Zhang, and S. Cheng. 2002. Solution properties of an $\alpha$-(1$\rightarrow$3)-D-glucan from Lentinus edodes and its sulfated derivatives. Carbohydr. Res. 337: 155-160   DOI   ScienceOn
19 Lowry, O. H., N. J. Rosebrough, L. Farr, and R. J. Randall. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 265-275
20 Waeghe, T., A. Darvill, M. Mcnell, and P. Albershim. 1983. Determanation, by methylation analysis, of the glycosyllinkage compositions of microgram quantities of complex carbohydrates. Carbohydr. Res. 123: 281-304   DOI   ScienceOn
21 Kim, S. W., H. J. Hwang, J. P. Park, C. H. Song, and J. W. Yun. 2002. Mycelial growth and exo-biopolymer production by submerged culture of various edible mushrooms under different media. Lett. Appl. Microbiol. 34: 56-61   DOI   ScienceOn
22 Carbone, I. and L. M. Kohn. 1993. Ribosomal DNA sequence divergence within internal transcribed spacer 1 of the Selerotiniaccae. Mycologia 85: 415-427   DOI
23 Xu, C. P. and J. W. Yun. 2004. Influence of aeration on the production and the quality of the exopolysaccharides from Paecilomyces tenuipes C240 in a stirred-tank fermenter. Enzyme Microb. Technol. 35: 33-39   DOI   ScienceOn
24 Lucey, J. A., M. Srimivasan, H. Singh, P. A. Munro. 2000. Characterization of commercial and experimental sodium caseinates by multiangle laser light scattering and sizeexclusion chromatography. J. Agric. Food Chem. 48: 1610-1616   DOI   ScienceOn
25 Hwang, H. J., S. W. Kim, C. P. Xu, J. W. Choi, C. W. Kim, and J. W. Yun. 2003. Production and molecular characteristics of four groups of exopolysaccharides from submerged culture of Phellinus gilvus. J. Appl. Microbiol. 94: 708-719   DOI   ScienceOn
26 Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalies and weight matrix choice. Nucleic Acids Res. 22: 4673-4680   DOI   ScienceOn
27 Kim, G. Y., H. S. Park, B. H. Nam, S. J. Lee, and J. D. Lee. 2003. Purification and characterization of acidic proteoheteroglycan from the fruiting body of Phellinus linteus (Berk. & M.A. Curtis) Teng. Bioresource Technol. 89: 81-87   DOI   ScienceOn
28 Peng, Y., L. Zhang, F. Zeng, and Y. Xu. 2003. Structure and antitumor activity of extracellular polysaccharides from mycelium. Carbohydr. Polym. 54: 297-303   DOI   ScienceOn
29 Sweet, D. P., R. H. Shapiro, and P. Albersheim. 1975. Quantitative analysis by various G.L.C. response-factor theories for partially methylated and partially ethylated alditol acetates. Carbohydr. Res. 40: 217-225   DOI   ScienceOn
30 Bao, X., C. Liu, J. Fang, and X. Li. 2001. Structural and immunological studies of a major polysaccharide from spores of Ganoderma lucidum (Fr.) Karst. Carbohydr. Res. 332: 67-74   DOI   ScienceOn
31 Theresa A. B. and J. B. Greg. 2003. A review of the nontarget effects of fungi used to biologically control plant diseases. Agric. Ecosyst. Environ. 100: 3-16   DOI   ScienceOn
32 Domer, J. E. and R. E. Garner. 1991. Fungal wall components and immunostimulation, pp 157-167. In Latge, J. P. and Boucias, D. (eds.). Fungal Cell Wall and Immune Response. Springer-Verlag; Berlin
33 Jin, Y., L. Zhang, L. Chen, Y. Chen, P. C. K. Cheung, and L. Chen. 2003. Effect of culture media on the chemical and physical characteristics of polysaccharides isolated from Poria cocos mycelia. Carbohydr. Res. 338: 1507-1515   DOI   ScienceOn
34 Fedonenko, Y. P., O. N. Konnova, G. V. Zatonsky, A. S. Shashkov, S. A. Konnova, E. L. Zdorovenko, V. V. Ignatov, and Y. A. Knirel. 2004. Structural of the O-polysaccharide of the lipopolysaccharide of Azospirillum irakense KBC1. Carbohydr. Res. 339: 1813-1816
35 Gutierrez, A., A. Prieto, and A. T. Martínez. 1996. Structural characterization of extracellular polysaccharides produced by fungi from the genus Pleurotus. Carbohydr. Res. 281: 143-154   DOI   ScienceOn
36 Sone, Y., R. Okuda, N. Wada, E. Kishida, and A. Misaki. 1985. Structures and antitumor activities of the polysaccharides isolated fruiting body and the growing culture of mycelium of Ganoderma lucidum. Agric. Biol. Chem. 49: 2641-2653   DOI
37 Kim, S. W., C. P. Xu, H. J. Hwang, J. W. Choi, C. W. Kim, and J. W. Yun. 2003. Production and characterization of exopolysaccharides from an enthomopathogenic fungus Cordyceps militaris NG3. Biotechnol. Prog. 19: 428-435   DOI   ScienceOn
38 Whiteny, S. E. C., J. E. Brigham, A. H. Darke, J. S. G. Reid, and M. J. Gidley. 1998. Structural aspects of the interaction of mannan-based polysaccharides with bacterial cellulose. Carbohydr. Res. 307: 299-309   DOI   ScienceOn
39 Bohn, J. A. and J. N. BeMiller. 1995. (1$\rightarrow$3)-$\beta$-D-glucans as biological response modifiers: A review of structurefunctional activity relationships. Carbohydr. Polym. 28: 3-14   DOI   ScienceOn
40 Park, Y. H., E. K. Moon, Y. K. Shin, M. A. Bae, J. K. Kim, and Y. H. Kim. 2000. Antitumor activity of Paecilimyces japonica is mediated by apoptotic cell death. J. Microbiol. Biotechnol. 10: 16-20