Cloning, nucleotide sequence, and transcriptional analysis of Aspergillus aculeatus no. F-50 cellobiohydrolase I (cbhI) gene

Journal of Fermentation and Bioengineering Pub Date : 1998-01-01 DOI:10.1016/S0922-338X(97)80345-5

Goro Takada, Takashi Kawaguchi, Jun-Ichi Sumitani, Motoo Arai

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引用次数: 22

Abstract

The cbhI gene, coding for a major cellobiohydrolase (CBHI) of Aspergillus aculeatus, was cloned and sequenced. The gene consists of 1620-bp and encodes a protein containing 540 amino acids with a calculated molecular mass of 56,723 Da. CBHI, composed of an N-terminal catalytic domain belonging to family 7 of the glycosyl hydrolases, and a C-terminal cellulose-binding domain (CBD) belonging to family I of the CBDs, showed high similarity with other fungal CBHIs, especially with that of Penicillium janthinellum. The cbhI gene transcription start points in A. aculeatus were defined by primer extension, and the putative promoter sequence was analyzed. This sequence was found to be closely related to the consensus sequences of various fungal genes. Transcription analysis by ribonuclease protection assay revealed that the cbhI gene is induced by low-molecular-weight cellooligosaccharide and repressed by glucose. The results emphasize the possibility that in the A. aculeatus cellulase system, cellobiose is the true inducer and the role of the cbhI gene lies within the cascade regulating cellulase induction.

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刺曲霉的克隆、核苷酸序列及转录分析。F-50纤维生物水解酶I (cbhI)基因

克隆并测序了针状曲霉(Aspergillus aculeatus)一种主要纤维生物水解酶(cbhI)的基因。该基因由1620 bp组成，编码一种含有540个氨基酸的蛋白质，计算出的分子质量为56,723 Da。cbi由糖基水解酶家族7的n端催化结构域和家族I的c端纤维素结合结构域(CBD)组成，与其他真菌的CBHIs具有高度的相似性，尤其是与青霉菌(Penicillium janthinellum)。通过引物延伸确定了针叶鱼cbi基因的转录起始点，并对可能的启动子序列进行了分析。该序列与多种真菌基因的共识序列密切相关。核糖核酸酶保护实验转录分析表明，cbi基因受低分子低聚糖诱导，受葡萄糖抑制。这些结果强调了在刺草纤维素酶系统中，纤维素二糖是真正的诱导剂的可能性，而cbhI基因的作用在于级联调节纤维素酶的诱导。

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