Molecular cloning and determination of the nucleotide sequence of a gene encoding an acid-stable alpha-amylase from Aspergillus kawachii
Molecular cloning and determination of the nucleotide sequence of a gene encoding an acid-stable alpha-amylase from Aspergillus kawachii
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DOI：
10.1016/0922-338x(96)80579-4
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发表时间：
1996-01-01
期刊：
影响因子：
--
通讯作者：
Oba, T
中科院分区：
文献类型：
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作者：
Kaneko, A;Sudo, S;Oba, T
从白曲霉 IFO4308 中克隆了编码酸稳定 α-淀粉酶 (asAA) 的互补 DNA 和基因组 DNA，并测定了它们的核苷酸序列。结构基因 (asaA) 由一个 1,920 bp 的开放阅读框组成，编码 640 个氨基酸。该基因由9个外显子和8个内含子组成。信号肽由21个氨基酸残基组成。从N端到第479位残基的氨基酸序列与黑曲霉酸性α-淀粉酶具有97％的同源性，从第480位残基到C端的氨基酸序列，包括生淀粉亲和位点，即TS区，其氨基酸序列富含苏氨酸和丝氨酸，是asAA的特征。携带该cDNA的酵母转化体产生并分泌能够消化生淀粉的酸稳定的α-淀粉酶。
Complementary and genomic DNAs encoding an acid-stable alpha-amylase (asAA) were cloned from Aspergillus kawachii IFO4308 and their nucleotide sequences were determined. The structural gene (asaA) consisted of a 1,920 bp open reading frame that encoded 640 amino acids. The gene consisted of 9 exons and 8 introns. The signal peptide was composed of 21 amino acid residues. The amino acid sequence from the N-terminus to the 479th residue showed 97% homology with the Aspergillus niger acid alpha-amylase and that from the 480th residue to the C-terminus, including the raw-starch-affinity site, i.e., the TS region, the amino acid sequence of which was rich in threonine and serine, was characteristic of the asAA. A yeast transformant carrying the cDNA produced and secreted an acid-stable alpha-amylase which was capable of digesting raw starch.