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

作　　者：韩晓燕[1] 包郁明[1] 辛凤姣[1] 黄颖[1] Christophe BLECKER 戴小枫[1] HAN Xiaoyan;BAO Yuming;XIN Fengjiao;HUANG Ying;DAI Xiaofeng(Key Laboratory of Agro-Products Processing, Ministry of Agriculture, Institute of Food Science and Technology,Chinese Academy of Agricultural Sciences, Beijing 100193, China;Department of Food Science, University of Liège, Liège 999013, Belgium)

机构地区：[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　　处：《食品科学》2017年第11期122-127,共6页Food Science

基　　金：科技部国际科技合作项目(2016-X31)

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)。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 peptideswere sequenced by electrospray ionization-tandem mass spectrometry(ESI-MS/MS).During the purification process,CPI hydrolysates were separated into three fractions:U-Ⅰ,U-Ⅲand U-III by UF and the obtained U-Ⅲ,with higherantibacterial ability,was further separated into QF-Ⅰ,QF-Ⅱand QF-Ⅲby AEC.After filtration,QF-Ⅱ,which showedhigher antibacterial ability,was further fractionated using semi-P-HPLC into four subfractions:PF-Ⅰ,PF-Ⅱ,PF-ⅢandPF-Ⅳ,among which,PF-Ⅲwas determined to have the highest antibacterial activity.The purity of PF-Ⅲshowed a singlepeak 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).

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 

分 类 号：TS201.3[轻工技术与工程—食品科学]