肼解法的改进及在测定多肽链碳末端氨基酸的应用

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中图分类号：Q517 文献标志码：A

Improvement of Hydrazinolysis Method and Its Application in Identification of Carbon-terminal Amino Acids of Polypeptide Chain

HAN Wenyu1， LI Kai2 ，WEI Xu⋅1 ， LI Baorui 3 ，WANG Yuanxiu1 （1.School of Biological Science and Technology，University of Jinan，Jinan 25oo22，Shandong，China; 2.Shandong Fupai Ejiao Co.，Ltd.，Jinan 25O4OO，Shandong，China； 3.Shandong Academy of Agricultural Sciences，Jinan 25O1oo，Shandong，China）

Abstract：Tosolvetheproblem that thecarbon-terminalaminoacidsweredificulttoseparateduringtheoperationof the traditionalhydrazinehydrolysismethod，animproved methodforthedeterminationof carbon-terminalaminoacids was proposed basedon the traditional hydrazine hydrolysis method.Hydrazine hydrazide hydrate was used to hydrolyze casein digests with diferent degrees of hydrolysis，and amino acid hydrazide was removed byusing heptanal and benzaldehyde， respectively，and carbon-terminal amino acidsobtainedbyboth methods were determinedbythin-layer chromatography and high-performance liquid chromatography with a t -test.The results show that the carbon-terminal amino acid solutions obtained by using heptanal and benzaldehyde to remove amino acid hydrazide contain lysine andornithine，and the amino acid content increases with the increase of hydrolysis degree，and there is no significant difference （ p>0.05 ）in the contentof lysineandornithineobtainedbythediferentmethods，verifyingthefeasibilityoftheimprovedhydrazide hydrolysis method，and determining the directionalenzymeenzymatic digestion of trypsinonargininesandlysines in tyrosine protein.

Keywords： hydrazolysis；casein； carbon-terminal amino acid；trypsin；directed enzymatic hydrolysis

食物蛋白是活性肽的主要来源，适宜的酶解条件可以有效地将其中的生物活性肽释放出来。（剩余6940字）