沙棘籽粕蛋白肽的稳定性及分离纯化

摘要/Abstract

摘要： 近年来，肥胖症患者人数逐年增加，预防和治疗肥胖成为迫切需要解决的社会问题。目前市面常见的减肥产品效果不佳，且部分药品副作用较多，因此探寻一种食源类抗肥胖产品具有重要意义。前期研究表明，来源于沙棘籽粕蛋白的酶解产物具有初步抗肥胖效果。本文以沙棘籽粕蛋白为原料，酶解制备具有抑制胰脂肪酶活性的多肽，通过超滤、大孔树脂分离、超高压液相色谱-质谱联用和分子对接对沙棘籽粕蛋白肽（SSPA）进行分离纯化、结构鉴定和筛选。以猪胰脂肪酶（PPL）抑制率为指标，稳定性分析表明，SSPA具有热稳定性和pH稳定性，适量Na+和Mg2+可显著提高SSPA的PPL抑制活性，SSPA在短期贮藏过程中不会因暴露于空气而影响PPL抑制活性。UPLC-MS/MS鉴定得到31种多肽，结合分子对接筛选得到6个肽段，小于3 kDa组分中上述寡肽含量分别为VR（2.90%）、FR（7.40%）、RDR（1.10%）、APYR（1.50%）、NLLHR（1.40%）和EEAASLR（1.10%）。固相合成测定其IC50值分别为VR（371.07 μg/mL）、FR（243.07 μg/mL）、RDR（250.50 μg/mL）、APYR（350.41 μg/mL）、NLLHR（220.70 μg/mL）、EEAASLR（510.55 μg/mL）。分子对接表明以上多肽通过氢键、π-π相互堆积与PPL结合。Pearson相关性分析表明，分子对接结合能高低与沙棘籽粕蛋白肽的PPL抑制率强弱之间存在显著正相关（r=0.865，p<0.05）。

关键词: 沙棘籽粕蛋白肽, 胰脂肪酶抑制率, 稳定性, 分离, 分子对接

Abstract: The number of obese patients was increasing recently. At present, the common anti-obesity products in the market were less effective and parts of drugs even caused side effects. Therefore, it was of great significance to explore food-derived anti-obesity product. According to the previous result, the seabuckthorn seed protein (SSP) hydrolysates possessed anti-obesity ability. In this study, the peptides were purified by ultrafiltration, macroporous resin separation, combining with autodock and ultra-high pressure liquid chromatography mass spectrometry to identify and screen. SSPA showed higher thermal stability and pH stability. Moreover, the PPL inhibitory rate of SSPA could be significantly improved by adding appropriate amount of Na+ and Mg2+, and which couldn’t be influenced by exposure to air during short-term storage. The component with molecular weight less than 3 kDa was obtained by ultrafiltration, then it was separated combined with dynamic adsorption by HPD826 resin. UPLC-MS/MS analysis revealed 31 peptides in SSPA, and six peptides with higher peptides content were predicted via molecular docking as anti-obesity peptides. The peptides (<3 kDa peptides part) content was as follows: VR (2.90%), FR (7.40%), RDR (1.10%), APYR (1.50%), NLLHR (1.40%) and EEAASLR (1.10%), respectively. The IC50 value of PPL inhibition rate was as follows: VR (371.07 μg/mL), FR (243.07 μg/mL), RDR (250.50 μg/mL), APYR (350.41 μg/mL), NLLHR (220.70 μg/mL), EEAASLR (510.55 μg/mL), respectively. The PPL-inhibiting actions of these peptides involved hydrogen bonding and π-π stacking. Pearson correlation analysis showed that there was a positive correlation between molecular binding energy and PPL inhibition rate of peptides from seabuckthorn seed protein (r=0.865, p<0.05).

Key words: peptide from seabuckthorn seed protein, porcine pancreatic lipase inhibitory rate, stability, separation, molecular docking

中图分类号:

TS209

相欢崔春. 沙棘籽粕蛋白肽的稳定性及分离纯化[J]. 食品科学, 0, (): 0-0.

Huan XIANG. Study on Stability and Sepration of Peptide from Seabuckthorn Seed Protein[J]. FOOD SCIENCE, 0, (): 0-0.

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