Combination of Derivatization-HPLC-MS and Enzymatic Hydrolysis-Edman Degradation for Amino Acid Sequence and Configuration of Polymyxin B Components

doi:10.1007/s10337-021-04091-2

	Combination of Derivatization-HPLC-MS and Enzymatic Hydrolysis-Edman Degradation for Amino Acid Sequence and Configuration of Polymyxin B Components
	Zhang, Hanzhi 1,2; Li, Feng 3; Dun, Junling 4; Sun, Ning5 ; Liu, Hao 1; Chen, Guiliang 6
	2021-11
发表期刊	CHROMATOGRAPHIA (IF:1.2[JCR-2023],1.7[5-Year])
ISSN	0009-5893
EISSN	1612-1112
卷号	84 期号:11 页码:1057-1064
DOI	10.1007/s10337-021-04091-2
摘要	An analysis strategy that can simultaneously determine the amino acid site information and configuration for polymyxin (PM) components is reported in this study. The main components of PMB1, PMB2, and PMB1-I (the Leucine at position 7 changed to be Isoleucine) were purified from the mixture of PMB. The PMB component was hydrolyzed by hydrochloric acid to be a mixture of amino acids, which were derivatized with Marfey's reagent of N-alpha-(5-fluoro-2,4-dinitrophenyl)-l-alanine amide (l-FDAA), and then, the configurations of amino acids were easily determined by HPLC-MS. However, the aforementioned method could not offer the information of amino acid at each site, a specific enzymatic hydrolysis method combined Edman degradation was developed. The fatty acyl-diaminobutyric acid (position 1) of PMB component was removed by ficin, and the PMB components were transformed to be PMB nonapeptide. The amino acids at each site of the nonapeptide were determined by N-terminal sequencing. Combining the results of the above methods, the amino acid information and configuration of each site in PM components can be determined, especially the l-Leucine and l-Isoleucine in PMB1 and PMB1-I. The workflow was useful for other PM analogues to identify the sequence and configuration for quality control.
关键词	Polymyxin B Amino acid sequence and configuration Derivatization with Marfey's reagent Enzymatic hydrolysis Edman degradation
收录类别	SCIE
语种	英语
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS记录号	WOS:000695763600002
出版者	SPRINGER HEIDELBERG
原始文献类型	Article; Early Access
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/128230
专题	免疫化学研究所_特聘教授组_抗体化学实验室
通讯作者	Zhang, Hanzhi; Chen, Guiliang
作者单位	1.Shanghai Inst Food & Drug Control, Dept Antibiot & Microorganisms, Shanghai 201203, Peoples R China; 2.Abiochem Biotechnol Co Ltd, Shanghai 200003, Peoples R China; 3.Xian Univ, Sch Chem Engn, Xian 710065, Peoples R China; 4.Shimadzu China Co Ltd, Shanghai 200233, Peoples R China; 5.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai 201210, Peoples R China; 6.Shanghai Ctr Drug Evaluat & Inspect, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Hanzhi,Li, Feng,Dun, Junling,et al. Combination of Derivatization-HPLC-MS and Enzymatic Hydrolysis-Edman Degradation for Amino Acid Sequence and Configuration of Polymyxin B Components[J]. CHROMATOGRAPHIA,2021,84(11):1057-1064.
APA	Zhang, Hanzhi,Li, Feng,Dun, Junling,Sun, Ning,Liu, Hao,&Chen, Guiliang.(2021).Combination of Derivatization-HPLC-MS and Enzymatic Hydrolysis-Edman Degradation for Amino Acid Sequence and Configuration of Polymyxin B Components.CHROMATOGRAPHIA,84(11),1057-1064.
MLA	Zhang, Hanzhi,et al."Combination of Derivatization-HPLC-MS and Enzymatic Hydrolysis-Edman Degradation for Amino Acid Sequence and Configuration of Polymyxin B Components".CHROMATOGRAPHIA 84.11(2021):1057-1064.