泡盛曲霉KT-11中可消化生淀粉的α-淀粉酶的分子克隆及核苷酸序列测定

Molecular cloning and determination of the nucleotide sequence of raw starch digesting alpha-amylase from Aspergillus awamori KT-11.

作者信息

Matsubara Takayoshi, Ben Ammar Youssef, Anindyawati Trisanti, Yamamoto Satoru, Ito Kazuo, Iizuka Masaru, Minamiura Noshi

机构信息

Laboratory of Enzyme Chemistry, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan.

出版信息

J Biochem Mol Biol. 2004 Jul 31;37(4):429-38. doi: 10.5483/bmbrep.2004.37.4.429.

DOI:10.5483/bmbrep.2004.37.4.429

PMID:15469730

Abstract

Complementary DNAs encoding alpha-amylases (Amyl I, Amyl III) and glucoamylase (GA I) were cloned from Aspergillus awamori KT-11 and their nucleotide sequences were determined. The sequence of Amyl III that was a raw starch digesting alpha-amylase was found to consist of a 1,902 bp open reading frame encoding 634 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. On the other hand, the sequence of Amyl I, which cannot act on raw starch, consisted of a 1,500 bp ORF encoding 499 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. The sequence of GA I consisted of a 1,920 bp ORF that encoded 639 amino acids. The signal peptide was composed of 24 amino acids. The amino acid sequence of Amyl III from the N-terminus to the amino acid number 499 showed 63.3% homology with Amyl I. However, the amino acid sequence from the amino acid number 501 to C-terminus, including the raw-starch-affinity site and the TS region rich in threonine and serine, showed 66.9% homology with GA I.

摘要

从泡盛曲霉KT - 11中克隆出编码α - 淀粉酶（淀粉酶I、淀粉酶III）和糖化酶（糖化酶I）的互补DNA，并测定了它们的核苷酸序列。发现可消化生淀粉的α - 淀粉酶淀粉酶III的序列由一个1902 bp的开放阅读框组成，编码634个氨基酸。该酶的信号肽由21个氨基酸组成。另一方面，不能作用于生淀粉的淀粉酶I的序列由一个1500 bp的开放阅读框组成，编码499个氨基酸。该酶的信号肽由21个氨基酸组成。糖化酶I的序列由一个1920 bp的开放阅读框组成，编码639个氨基酸。信号肽由24个氨基酸组成。从N端到第499位氨基酸的淀粉酶III的氨基酸序列与淀粉酶I的同源性为63.3%。然而，从第501位氨基酸到C端的氨基酸序列，包括生淀粉亲和位点和富含苏氨酸和丝氨酸的TS区域，与糖化酶I的同源性为66.9%。