皖浙花猪干腌火腿源血糖调节肽的分离纯化、鉴定及量子化学表征  

Isolation,Purification,Identification and Quantum Chemical Characterization of Blood Glucose-Regulating Peptides Derived from Dry-Cured Ham of Wanzhe Spotted Pigs

作　　者：黄晶晶[1,2] 周迎芹 罗章 刘振东 程秀峰 谢宁宁 HUANG Jingjing;ZHOU Yingqin;LUO Zhang;LIU Zhendong;CHENG Xiufeng;XIE Ningning(Institute of Agro-product Science and Technology,Anhui Academy of Agricultural Sciences,Hefei 230001,China;Anhui Engineering Laboratory of Food Microbial Fermentation and Functional Application,Hefei 230031,China;College of Food Science,Tibet Agriculture and Animal Husbandry University,Nyingchi 860000,China)

机构地区：[1]安徽省农业科学院农产品加工研究所,安徽合肥230001 [2]安徽省食品微生物发酵与功能应用工程实验室,安徽合肥230031 [3]西藏农牧学院食品科学学院,西藏林芝860000

出　　处：《食品科学》2024年第3期25-33,共9页Food Science

基　　金：安徽省自然科学基金青年项目(2108085QC145);安徽省重点研发科技援藏项目(2022j11020001);中央引导地方科技发展资金项目(XZ202202YD0004C);安徽省农业科学院平台项目(2023YL010,2023YL017)。

摘　　要：为探究小肽抑制糖类消化的机理,制备皖浙花猪干腌火腿肌肉的水提物和胃胰酶消化产物,分离纯化出α-淀粉酶和α-葡萄糖苷酶抑制活性较高的组分,鉴定、筛选其中的肽序列,采用量子化学方法计算分子前线轨道分布和能量、静电荷分布、键长等结构和电荷参数,推测活性位点。结果表明:1)酶解后火腿肌肉的粒径减小、降糖活性提高;2)水提物和胃胰酶消化产物经过葡聚糖凝胶分离后分别获得两个组分(S-Ⅰ、S-Ⅱ)和3个组分(WY-Ⅰ、WY-Ⅱ和WY-Ⅲ);3)利用质谱技术从高活性组分WY-Ⅱ中鉴定出104条长度8~24的肽序列,筛选出Peptide Ranker程序评分大于0.7的5条序列;4)前述序列的最高占据轨道多分布在精氨酸的胍基及氨基端附近基团,而最低未占轨道多分布在羧基端及附近基团;5)能级差ΔEL-H较低的序列GPMGPSGPR、LGFGGPSGPNAGR、APAPAPAPAPAPPK可能具有较高活性。根据库伦定律,其活性位点分别定位于精氨酸的C106H108、亮氨酸的C10H12和赖氨酸的C176H177,都位于C—H键。研究为探究肽的血糖调节机制,证明地方品种猪的营养价值提供了理论支撑。To investigate the inhibitory mechanism of small peptides on carbohydrate digestion,α-amylase andα-glucosidase inhibitory fractions from the water extract and the gastropancreatic digest of dry-cured ham muscle of Wanzhe spotted pigs were separated,purified,identified,and screened for peptide sequences.And the quantum chemical calculation was used to calculate structural and charge parameters including the distribution and energy of the frontier orbitals,electrostatic charge distribution and bond length,in order to speculate the active sites.It was found that the particle size of ham muscle decreased and its hypoglycemic activity increased after proteolysis.Two(S-I and S-II)and three fractions(WY-I,WY-II and WY-III)were obtained from the water extract and the digest after Sephadex column chromatography,respectively.Using mass spectrometry,104 peptide sequences consisting of 8–24 amino acids were identified from fraction WY-II andfive sequences with Peptide Ranker scores greater than 0.7 were selected.The highest occupied orbitals of thefive sequences were mainly distributed in the guanidine groups of arginine and the groups close to the amino-terminal end,while the lowest unoccupied orbitals were in the carboxyl terminals and nearby groups.Sequences with lowerΔEL-H values,GPMGPSGPR,LGFGGPSGPNAGR and APAPAPAPAPPK,might be more active.According to Coulomb’s law,the active sites of these three peptides were located at–C106H108 of arginine,–C10H12 of leucine and–C176H177 of lysine,respectively.This study could provide theoretical support for understanding the blood glucose-regulating mechanism of peptides and the nutritional value of local pig breeds.

关 键 词：血糖调节 α-淀粉酶和α-葡萄糖苷酶抑制活性 肽序列 量子化学计算 干腌火腿 

分 类 号：TS201.4[轻工技术与工程—食品科学]