| Project/Area Number |
18580332
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Boundary agriculture
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| Research Institution | University of Miyazaki |
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Principal Investigator |
OHTA Kazuyoshi University of Miyazaki, Department of Biochemistry and Applied Biosciences, Professor (70112315)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Sachio UNIVERSITY OF MIYAZAKI, Department of Applied Chemistry, Professor (90148916)
HUROSE Jyun UNIVERSITY OF MIYAZAKI, Department of Applied Chemistry, Associate Professor (60264363)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2006: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | Penicillium citrinum / Aspergillus japonicus / xylan / xylanase / β-xylosidase / filamentous fungi / biomass / ethanol / xylan / xylanase / β-xylosidase / filamentous fungi / biomass / ethanol / 発酵 / 大腸菌 |
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| Research Abstract |
1)An extracellular endo-1,4-β-xylanase was purified from the culture filtrate of a filamentous fungus, Penicillium citrinum strain FERM P-15944. The purified enzyme showed a single band on SDS-PAGE with an apparent M_r of 31.6 kDa. An open reading frame of the xylanase gene ^<xynB was interrupted by nine introns and encoded a presumed prepropeptide of 25 amino acids and a mature protein of 302 amino acids. Sequence alignment showed that the P. citrinum enzyme belongs to glycoside hydrolase family 10.
2)An extracellular protein exhibiting β-xylosidase activity was purified from the culture filtrate of a filamentous fungus Aspergillus japonicus strain MU-2. The purified enzyme was a glycoprotein with an apparent M_r of 113.2 kDa The enzyme also had hydrolytic activities toward p-nitropheny1-β-D-glucopyranoside and p-nitropheny1-β-L-arabinofuranoside. An open reading frame of the β-xylo, sidase gene ^<xyl>A, consisting of 2412 bp, was not interrupted by introns, and it encoded a presumed s
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ignal peptide of 17 amino acids and a mature protein of 787 amino acids. The deduced amino acid sequence showed a high degree of identity (69%)to Aspergillus nigerβ-xylosidase XlnD that belongs to glycoside hydrolase family 3.
3)An extracellular xylanase from A. japonicus was purified with a yield of 28.7% of the activity and a 21-fold increase in specific activity. The xylanase showed a single band on SDS-PAGE with an apparent M_r of 25.1 kDa. A pair of degenerate PCR primers was synthesized according to the internal amino acid sequence (EDYGEYN)and a highly conserved sequence (NHFNAWA)among GH family-11 xylanases from Aspergillus spp. Two distinct internal sequences were amplified from genomic DNA of A. japonicus as a template by PCR with the primer pair. Two genomic DNA segments encoding xylanase were cloned using two DIG-labeled amplified fragments The deduced amino acid sequences of two xylanase genes, ^<xynG1 and ^<xynG2, revealed that purified xylanase corresponded to ^<xynG2 gene product. Less
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