裂褶菌Schizophyllum commune之葡聚糖裂解酵素(β-1,3-glucanase) 之基因選殖及其生化活性探討

吳秀芳

doi:10.6342/NTU.2004.02209

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裂褶菌Schizophyllum commune之葡聚糖裂解酵素(β-1,3-glucanase) 之基因選殖及其生化活性探討

Cloning and characterization of β-1,3-glucanase gene from mycoparasitic Schizophyllum commune

吳秀芳 , Masters　　Advisor：曾顯雄　　

繁體中文 DOI： 10.6342/NTU.2004.02209

gene cloning ； 3-glucanase ； biocontrol ； Schizophyllum commune ； mycoparasitism ； β-1 ； 3-glucanase ； β-1 ； Schizophyllum commune ； mycoparasitism biocontrol ； gene cloning

Abstract │ Reference (50) │ Altmetrics

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Schizophyllum commune was enhanced to produce higher yield ofβ-1,3-glucanasewhile the basal medium was amended with the cell wall of  Rhizoctonia solani.β-1,3-glucanasewas partially purified by anion-exchange, hydrophobic-interaction chromatography, which increase 251.5 fold purtification and 13,579 specific activity. The pI of the enzyme was pH 3.45. The partially purified endo-β-1,3-glucanase was assumed to be a tetraisomer, consisting of 63kDa、58 kDa、29 kDa and 27 kDa monomer, respectively, as shown by SDS-PAGE. The N-terminal amino acid residues of 63 kDa and 58 kDa protein were LDNGVGAL and LDNGVGRL, respectively by Edman degradation. By different combination of the degenerate primers from N-terminal amino acid residues plus 7 degenerate primers from C-terminal conservative region to process RT-PCR, a 500 bp cDNA product was obtained, which shows identical N-terminal sequence as the original primers, but however also contained unexpected stop codon . The cDNA sequence will be used to probe the newly constructed Fosmid library to access the possibly hidden β-1,3-glucanasegene.

Reference ( 50 ) 〈TOP〉

Bara, M. T. F., Lima, A. L., and Ulhoa, C. J. 2003. Purification and characterization of an exo-β-1,3-glucanase produced by Trichoderma harzianum. FEMS Microbiology Letters 219:81-85.
連結：
Barnett, H. L. 1964. Mycoparasitism. Mycologia. 56:1-19.
連結：
Barnett, H. L., and Binder, F. L. 1973. The fungal host-parasite relationship. Annu. Rev. Phytopathol. 11:273-292.
連結：
Benhamou, N., and Chet, I. 1997. Cellular and molecular mechanisms involved in the interaction between Trichoderma harzianum and Pythium ultimum. Appl. Environ. Microbiol. 63:2095-2099.
連結：
Broglie, K., Chet, I., Holliday, M., Cressman, R., Biddle, P., Knowlton, S., Mauvais, C. J., and Broglie, R. 1991. Transgentic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254:1194-1197.
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