棉籽抗菌活性肽的分离纯化及鉴定

doi:10.7506/spkx1002-6630-201711020

食品科学 ›› 2017, Vol. 38 ›› Issue (11): 122-127.doi: 10.7506/spkx1002-6630-201711020

棉籽抗菌活性肽的分离纯化及鉴定

韩晓燕，包郁明，辛凤姣，黄颖，Christophe BLECKER，戴小枫

1.中国农业科学院农产品加工研究所，农业部农产品加工重点实验室，北京 100193；2.列日大学食品学院，比利时列日 999013

出版日期:2017-06-15 发布日期:2017-06-19

Purification and Identification of Antibacterial Peptides from Cottonseed Protein Isolate Hydrolysates

HAN Xiaoyan, BAO Yuming, XIN Fengjiao, HUANG Ying, Christophe BLECKER, DAI Xiaofeng

1. Key Laboratory of Agro-Products Processing, Ministry of Agriculture, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. Department of Food Science, University of Liège, Liège 999013, Belgium

Online:2017-06-15 Published:2017-06-19

摘要/Abstract

摘要： 通过体外模拟消化系统对棉籽分离蛋白（cottonseed protein isolate，CPI）进行酶解，得到具有抗菌活性的酶解产物，并采用超滤（ultrafiltration，UF）、阴离子交换色谱（anion exchange chromatography，AEC）、半制备高效液相色谱（semi-preparation high performance liquid chromatography，semi-P-HPLC）分离技术对棉籽抗菌活性肽进行分离纯化，用电喷雾串联质谱（electrospray ionization-tandem mass spectrometry，ESI-MS/MS）鉴定棉籽抗菌肽的氨基酸序列。在抗菌活性肽分离纯化过程中，用UF对具有抗菌活性的CPI酶解产物进行分离，得到3 个组分U-Ⅰ～U-Ⅲ。抗菌活性检测表明U-Ⅲ的抗菌能力最强；用AEC分离U-Ⅲ得到3 个组分QF-Ⅰ～QF-Ⅲ，其中QF-Ⅱ抗菌能力最强；进一步采用semi-P-HPLC分离QF-Ⅱ得到4 个组分PF-Ⅰ～PF-Ⅳ，其中PF-Ⅲ的抗菌能力最强，经HPLC检测为单一峰，ESI-MS/MS检测分析得到该肽的氨基酸序列为ISGLIYEETR（Ile-Ser-Gly-Leu-Ile-Tyr-Glu-Glu-Thr-Arg）。

关键词: 棉籽分离蛋白酶解产物, 抗菌活性肽, 分离纯化, 氨基酸序列

Abstract: Cottonseed protein isolate (CPI) was digested in vitro to prepare hydrolysates with antibacterial activity. Ultrafiltration (UF), anion exchange chromatography (AEC), and semi preparative high performance liquid chromatography (semi-P-HPLC) were used to isolate and purify antibacterial peptides from CPI hydrolysates, and the purified peptides were sequenced by electrospray ionization-tandem mass spectrometry (ESI-MS/MS). During the purification process, CPI hydrolysates were separated into three fractions: U-Ⅰ, U-Ⅲ and U-III by UF and the obtained U-Ⅲ, with higher antibacterial ability, was further separated into QF-Ⅰ, QF-Ⅱ and QF-Ⅲ by AEC. After filtration, QF-Ⅱ, which showed higher antibacterial ability, was further fractionated using semi-P-HPLC into four subfractions: PF-Ⅰ, PF-Ⅱ, PF-Ⅲ and PF-Ⅳ, among which, PF-Ⅲ was determined to have the highest antibacterial activity. The purity of PF-Ⅲ showed a single peak in HPLC. Finally, by ESI-MS/MS, and the amino acid sequence of the peptide was identified as ISGLIYEETR (Ile-Ser-Gly-Leu-Ile-Tyr-Glu-Glu-Thr-Arg).

Key words: cottonseed protein isolate hydrolysates, peptide with antibacterial activity, isolation and purification, amino acid sequence

中图分类号:

TS201.3

韩晓燕，包郁明，辛凤姣，黄颖，Christophe BLECKER，戴小枫. 棉籽抗菌活性肽的分离纯化及鉴定[J]. 食品科学, 2017, 38(11): 122-127.

HAN Xiaoyan, BAO Yuming, XIN Fengjiao, HUANG Ying, Christophe BLECKER, DAI Xiaofeng. Purification and Identification of Antibacterial Peptides from Cottonseed Protein Isolate Hydrolysates[J]. FOOD SCIENCE, 2017, 38(11): 122-127.

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棉籽抗菌活性肽的分离纯化及鉴定

Purification and Identification of Antibacterial Peptides from Cottonseed Protein Isolate Hydrolysates

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