Molecular cloning and characterization of a cDNA encoding cellobiose dehydrogenase from the wood-rotting fungus Grifola frondosa.
Molecular cloning and characterization of a cDNA encoding cellobiose dehydrogenase from the wood-rotting fungus Grifola frondosa.
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编码来自腐木真菌 Grifola frondosa 的纤维二糖脱氢酶的 cDNA 的分子克隆和表征。
DOI：
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发表时间：
2002
影响因子：
2.1
通讯作者：
M. Samejima
中科院分区：
文献类型：
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作者：
Makoto Yoshida;T. Ohira;K. Igarashi;H. Nagasawa;M. Samejima
利用反转录-聚合酶链反应（RT-PCR）和cDNA末端快速扩增技术，从灰树花的木材腐朽菌灰树花中克隆了一个编码纤维二糖脱氢酶（CDH）的cDNA。CDH cDNA全长2469 bp，包括一个开放阅读框，编码N端18个氨基酸的信号肽和750个氨基酸的成熟蛋白，预测分子量为79.6 kDa，pI值为4.32。氨基酸序列分析表明，它包含一个黄素结合基序，两个葡萄糖-甲醇-胆碱氧化还原酶基序，和两个可能的残基血红素配体结合（Met 61和His 58）。G. frondosa CDH（GfrCDH）与3个已知的担子菌CDH具有高度的同源性，但与2个子囊菌CDH没有同源性。此外，CDH基因在G.通过RT-PCR分析在几种碳源上生长的frondosa。在纤维二糖和纤维素上生长的菌丝体中检测到GfrCDH的mRNA，但在葡萄糖上没有。因此，GfrCDH基因的转录似乎在有利于纤维素降解的条件下得到促进，并受到碳分解代谢产物阻遏的调节。
Cloning of a cDNA encoding cellobiose dehydrogenase (CDH) from the wood-rotting fungus Grifola frondosa, which produces the edible maitake mushroom, was performed using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The CDH cDNA consisted of 2469 bp, including an open reading frame encoding the 18-amino acid signal peptide at the N-terminal region and the 750-amino acid mature protein with a predicted molecular mass of 79.6 kDa and a pI value of 4.32. Analysis of the amino acid sequence revealed that it contains a flavin-binding motif, two glucose-methanol-choline oxidoreductase motifs, and two possible residues for heme ligand binding (Met61 and His58). The amino acid sequence of G. frondosa CDH (GfrCDH) has a high degree of identity with three known CDHs from basidiomycetes, but not with two CDHs from ascomycetes. In addition, transcription of the CDH gene in G. frondosa grown on several carbon sources was analyzed by RT-PCR. mRNA of GfrCDH was detected from mycelia grown on cellobiose and cellulose, but not on glucose. Consequently, transcription of the GfrCDH gene seems to be promoted under conditions favoring cellulose degradation, and to be regulated by carbon catabolite repression.