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一项关于从脱脂米糠中获得的脂肪酶抑制肽的研究。

An study of lipase inhibitory peptides obtained from de-oiled rice bran.

作者信息

Ketprayoon Titima, Noitang Sajee, Sangtanoo Papassara, Srimongkol Piroonporn, Saisavoey Tanatorn, Reamtong Onrapak, Choowongkomon Kiattawee, Karnchanatat Aphichart

机构信息

Program in Biotechnology, Faculty of Science, Chulalongkorn University 254 Phayathai Road, Pathumwan Bangkok 10330 Thailand.

Research Unit in Bioconversion/Bioseparation for Value-Added Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn University 254 Phayathai Road, Pathumwan Bangkok 10330 Thailand

出版信息

RSC Adv. 2021 May 25;11(31):18915-18929. doi: 10.1039/d1ra01411k. eCollection 2021 May 24.

Abstract

De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry. DORB may prove to be an important protein source, and also contains many other micronutrients. This study has the principal aim of optimizing the process of DORB protein hydrolysate preparation, and then testing the hydrolysate to determine its lipase inhibitory activity. DORB underwent hydrolysis using Alcalase® and response surface methodology (RSM). The resulting degree of hydrolysis (DH) was then monitored along with the extent of any lipase inhibitory activity. The optimum levels of lipase inhibition were obtained at a temperature of 49.88 °C, a duration of 150.43 minutes, and 1.53% Alcalase® used for the sample 5% (w/v) solution. In these conditions, the DH value was 35.65%, and the IC value for lipase inhibitory activity was 2.84 μg mL. Five ranges of different molecular weights were obtained fractionation, whereupon it was determined that the highest level of inhibitory activity was achieved by the <0.65 kDa fraction. This fraction was then further purified RP-HPLC, and the resulting peak had a retention time of 21.75 minutes ( sub-fraction) and exhibited high lipase inhibitory activity. Mass spectrometry was used to determine the amino acid sequence for this peak, identified as FYLGYCDY. This particular peptide is categorized as bitter, with a non-toxic profile, and having poor water solubility. The synthesized form of this peptide showed lipase inhibitory activity measured by an IC value of 0.47 ± 0.02 μM. The Lineweaver-Burk plot revealed that FYLGYCDY is a non-competitive inhibitor, while analysis of the docking results provided details of the FYLGYCDY peptide binding site with the porcine pancreatic lipase (PPL) complex, which is a competitive type. It can be inferred from these findings that DORB may prove a useful raw material source for the production of anti-obesity peptides which might enhance the therapeutic and commercial performance of functional foods and healthcare products.

摘要

脱脂米糠(DORB)是米糠油工业一种潜在有用的副产品。DORB可能被证明是一种重要的蛋白质来源,并且还含有许多其他微量营养素。本研究的主要目的是优化DORB蛋白水解物的制备工艺,然后测试该水解物以确定其脂肪酶抑制活性。使用碱性蛋白酶(Alcalase®)和响应面法(RSM)对DORB进行水解。然后监测所得的水解度(DH)以及任何脂肪酶抑制活性的程度。在温度为49.88℃、持续时间为150.43分钟以及用于5%(w/v)样品溶液的1.53%碱性蛋白酶(Alcalase®)的条件下获得了最佳脂肪酶抑制水平。在这些条件下,DH值为35.65%,脂肪酶抑制活性的IC值为2.84μg/mL。通过分级分离获得了五个不同分子量范围,据此确定<0.65 kDa级分实现了最高水平的抑制活性。然后通过反相高效液相色谱(RP-HPLC)对该级分进行进一步纯化,所得峰的保留时间为21.75分钟(亚级分),并表现出高脂肪酶抑制活性。使用质谱法确定该峰的氨基酸序列,鉴定为FYLGYCDY。这种特定的肽被归类为苦味肽,具有无毒特性,并且水溶性较差。该肽的合成形式显示出脂肪酶抑制活性,通过IC值0.47±0.02μM来衡量。Lineweaver-Burk图显示FYLGYCDY是一种非竞争性抑制剂,而对接结果分析提供了FYLGYCDY肽与猪胰脂肪酶(PPL)复合物结合位点的详细信息,这是一种竞争性类型。从这些发现可以推断,DORB可能被证明是生产抗肥胖肽的有用原料来源,这可能会提高功能性食品和保健品的治疗和商业性能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/28e5/9033478/cb72380d3956/d1ra01411k-f1.jpg

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