Mikami B, Nomura K, Morita Y
J Biochem. 1986 Aug;100(2):513-6. doi: 10.1093/oxfordjournals.jbchem.a121741.
The blocked N-terminus and N-terminal sequence of soybean beta-amylase were determined by analyzing the acidic peptides derived on peptic digestion of the enzyme. The acidic peptides were separated from the digest on a Dowex 50 X 2 column and purified by reversed phase-high performance liquid chromatography (RP-HPLC). The major acidic peptide, Pep-4, was a heptapeptide with a molecular weight of 766. Forty-eight hundredths mol acetyl group and 0.61 mol acetyl-Ala per mol of Pep-4 were detected on RP-HPLC analysis. The N-terminal 9 amino acid sequence of soybean beta-amylase was deduced to be acetyl-Ala-Thr-Ser-Asp-Ser-Asn-Met-(Gly-Leu) from the results of sequence analysis of Pep-4 and amino acid analysis of other acidic peptides.
通过分析大豆β-淀粉酶经胃蛋白酶消化产生的酸性肽段,确定了该酶的封闭N端和N端序列。酸性肽段在Dowex 50 X 2柱上从消化物中分离出来,并通过反相高效液相色谱(RP-HPLC)进行纯化。主要的酸性肽段Pep-4是一种七肽,分子量为766。RP-HPLC分析检测到每摩尔Pep-4含有0.48摩尔乙酰基和0.61摩尔乙酰丙氨酸。根据Pep-4的序列分析结果和其他酸性肽段的氨基酸分析结果,推断大豆β-淀粉酶的N端9个氨基酸序列为乙酰丙氨酸-苏氨酸-丝氨酸-天冬氨酸-丝氨酸-天冬酰胺-甲硫氨酸-(甘氨酸-亮氨酸)。