首页|蛋白核小球藻胰脂肪酶抑制肽的分离纯化、鉴定及其降脂活性

蛋白核小球藻胰脂肪酶抑制肽的分离纯化、鉴定及其降脂活性

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
  • 维普
以蛋白核小球藻为原料制备小球藻蛋白酶解物,采用超滤和葡聚糖凝胶对其进行分离纯化,以胰脂肪酶活性抑制率为指标筛选具有降脂活性的肽组分,获得蛋白核小球藻胰脂肪酶抑制肽(记为PES).通过测定高糖饮食秀丽隐杆线虫休内的脂肪沉积、甘油三酯和总胆固醇水平评价纯化肽PES的降脂活性.采用液相色谱-串联质谱鉴定PES肽序列,结合分子对接技术筛选出潜在的胰脂肪酶抑制肽段,进一步验证合成多肽的胰脂肪酶抑制活性.结果表明,PES具有较好的降脂活性,质量浓度为1mg/mL时能够抑制高糖饮食线虫体内22.5％的脂肪沉积,降低27.4％甘油三酯和29.4％总胆固醇水平;经鉴定得到999条肽段,进一步筛选得到4条潜在的胰脂肪酶抑制肽段,其中胰脂肪酶活性抑制率最高的肽段是FLGPF,在8mg/mL时抑制率达50.12％,抑制类型为可逆非竞争性抑制,抑制常数为5.23 mg/mL;分子对接表明FLGPF可与人胰脂肪酶以π-氢键、π-阳离子和氢键作用更好地结合.该研究可为蛋白核小球藻降脂肽的开发利用提供理论参考.
Isolation,Purification,Identification and Hypolipidemic Activity of Lipase Inhibitory Peptide from Chlorellapyrenoidosa
In this study,pancrelipase inhibitory peptides(PES)from an enzymatic protein hydrolysate of Chlorella pyrenoidosa were isolated and purified by ultrafiltration and Sephadex gel chromatography.The in vivo hypolipidemic activity of PES was evaluated by fat deposition and the levels of triglyceride(TG)and total cholesterol(TC)in Caenorhabditis elegans fed a high sugar diet.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was used to identify the peptide sequence of PES,and molecular docking was used to select potential pancreatic lipase inhibitory peptides,and the pancreatic lipase inhibitory activity of the synthesized peptides was verified.The results showed that PES had good hypolipidemic activity at a concentration of 1 mg/mL;it inhibited lipid deposition by 22.5％,and reduced the levels of TG and TC by 27.4％and 29.4％,respectively.In total,999 peptides were identified,and four potential lipase inhibitory peptides were obtained.Among them,FLGPF had the best inhibitory effect on pancreatic lipase,with an inhibition rate of 50.12％at 8 mg/mL.The inhibition was reversible and non-competitive,with an inhibition constant of 5.23 mg/mL.Molecular docking showed that FLGPF could better bind to human pancreatic triacylglycerol lipase(PTL)viaπ-hydrogen,π-cation and hydrogen bond interactions.This study can provide a theoretical reference for the development and utilization of C.pyrenoidosa protein-derived hypolipidemic peptide.

Chlorella pyrenoidosalipase inhibitory peptideCaenorhabditis eleganshypolipidemicmolecular dockinginhibitory kinetics

林娈、柳雯郡、黄俊媛、贾瑷菁、汪登谜、刘斌、赵超

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福建农林大学食品科学学院,福建福州 350002

泉州师范学院海洋与食品学院,福建泉州 362000

福建省海洋藻类活性物质制备与功能开发重点实验室,福建泉州 362000

蛋白核小球藻 胰脂肪酶抑制肽 秀丽隐杆线虫 降脂 分子对接 抑制动力学

福建省自然科学基金面上项目泉州市科技计划国家级大学生创新创业训练计划

2020J017872019N025201910399014

2023

10.7506/spkx1002-6630-20230302-021

食品科学
北京食品科学研究院

食品科学

CSTPCDCSCD北大核心
影响因子：1.327
ISSN：1002-6630
年,卷(期)：2023.44(24)
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