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Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus

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Summary

Wa have cloned and characterized the cDNA coding for a major component of cellulase, endoglucanase (FI-CMCase), produced by Aspergillus aculeatus. The cDNA was isolated from a A. aculeatus cDNA library using synthetic oligonuceotide mixtures that correspond to the internal amino acid sequence of the mature FI-CMCase protein. Nucleotide sequence analysis of the cloned cDNA insert revealed a 711 bp open reading frame that encoded a protein of 237 amino acid residues. The primary structure of FI-CMCase deduced from the nucleotide sequence of cDNA agreed with that found by amino acid sequencing of peptide fragments obtained by digestion with several proteinases and cyanogen bromide cleavage. There may be a signal peptide sequence of 16 amino acid residues at the N-terminus. The molecular mass of the mature protein calculated from the cDNA is 24002 daltons, which compares favorably with molecular mass estimates of purified FI-CMCase obtained from SDS-PAGE (25000 Da). No distinct homology was found between the amino acid sequence of FI-CMCase and known cellulase sequences of other microorganisms. This study is the first example of cDNA cloning of an endoglucanase from the genus Aspergillus.

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Communicated by M. Yanagida

The sequence reported in this paper has been submitted to the EMBL/GenBank through DDBJ (accession no, X52525).

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Ooi, T., Shinmyo, A., Okada, H. et al. Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus . Curr Genet 18, 217–222 (1990). https://doi.org/10.1007/BF00318384

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