Preparation and Activity Analysis of DPP-IV Inhibitory Peptides from Pacific Oyster (Crassostrea gigas)

Abstract

Abstract: Dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides were prepared from Pacific oyster (Crassostrea gigas). By comparing the inhibitory activities of the hydrolysates prepared using 5 kinds of proteinases, including papain, alcalase, pancreatin, neutrase and protromex on DPP-IV, the pancreatin hydrolysate revealed the highest inhibitory activity. Optimal hydrolyzing condition was determined to be with 0.8% pancreatin, at pH 8.0, 37?C and hydrolysis for 90 min. The hydrolysate was further purified by ultrafiltration, Sephadex G-15 and reversed-phase high performance liquid chromatography (RP-HPLC). After purification, 2 peptides were obtained and identified as Glu-Ile-Thr-Ala-Leu-Ala-Pro-Ser-Thr-Met-Lys (EITALAPSTMK) and Ile-Leu-Ala-Pro-Pro-Glu-Arg (ILAPPER) by mass spectrometry. The digestibility characteristics of these two peptides in gastrointestinal fluid were analyzed using online simulation of BIOPEP. Two peptides APSTM and ILAPPER were synthesized by solid-phase synthesis. The results showed that IC50 values of APSTM and ILAPPER were 354.81 ?mol/L and 16.98 ?mol/L, respectively. Using oyster as raw material, peptides with effective DPP-IV inhibitory activity were prepared, which may provide a solid basis for comprehensive utilization of oyster in the future.

Key words: oyster, hydrolysis, DPP-IV inhibitory peptide, isolation and purification, inhibitory activity

CLC Number:

TS254.1

References

[1] 农业部渔业渔政管理局.中国渔业统计年鉴[M]. 北京: 中国农业出版社, 2019. [2]李亚, 刘淇, 张娟娟, 等.不同前处理方式对牡蛎肽营养成分的影响[J].食品安全质量检测学报, 2018, 9(13):3298-3302 [3]李茹, 韦玉白, 高永利, 等.牡蛎酶解物中的新型抗氧化肽[J].食品科技, 2017, 42(8):19-25 [4]CHEONG K L, XIA L X, LIU Y.Isolation and characterization of polysaccharides from oysters (Crassostrea gigas) with anti-tumor activities using an aqueous two-phase system[J].Marine Drugs, 2017, 15(11):338-349 [5]邱娟, 沈建东, 翁凌, 等.利用牡蛎制备抑制肽的工艺优化[J].食品科学, 2017, 38(16):165-172 [6]邓嫣容.牡蛎复合酶解提高蚝油风味的工艺探讨[J].现代食品科技, 2011, 27(7):788-791 [7]米顺利, 高程海, 姜舒, 等.榄钱牡蛎软罐头的工艺研究[J].现代食品科技, 2018, 34(1):203-208 [8]孔繁东, 祖国仁, 丁琦, 等.牡蛎酱油的研究与开发[J].中国调味品, 2008, 33(7):34-37 [9]DRUCKER D J.The role of gut hormones in glucose homeostasis[J].The Journal of Clinical Investigation, 2007, 117(1):24-32 [10]DEACON C F, HOLST J J.Dipeptidyl peptidase IV inhibition as an approach to the treatment and prevention of type 2 diabetes: A historical perspective[J].Biochemical & Biophysical Research Communications, 2002, 294(1):1-4 [11]DEACON C F.Dipeptidyl peptidase-4 inhibitors in the treatment of type 2 diabetes: a comparative review[J].Diabetes, Obesity & Metabolism, 2011, 13(1):7-18 [12] LACROIX I M E, LI-CHAN E C Y.Food-derived dipeptidyl-peptidase IV inhibitors as a potential approach for glycemic regulation – Current knowledge and future research considerations[J]. Trends in Food Science & Technology, 2016, 54: 1-16. DOI:10.1016/j.tifs.2016.05.008. [13]LIU R, CHENG J, WU H.Discovery of food-derived dipeptidyl peptidase IV inhibitory peptides: a review[J].International Journal of Molecular Sciences, 2019, 20(3):1-22 [14]JANSEN J, KARGES W, RINK L.Zinc and diabetes--clinical links and molecular mechanisms[J].Journal of Nutritional Biochemistry, 2009, 20(6):399-417 [15]ZHANG Y, CHEN R, CHEN X, et al.Dipeptidyl peptidase IV-inhibitory peptides derived from silver carp (Hypophthalmichthys molitrix Val) proteins[J].Journal of Agricultural and Food Chemistry, 2016, 64(4):831-839 [16] SATO K, MIYASAKA S, TSUJI A, et al.Isolation and characterization of peptides with dipeptidyl peptidase IV (DPPIV) inhibitory activity from natto using DPPIV from Aspergillus oryzae[J]. Food Chemistry, 2018, 261: 51-56. DOI:10.1016/j.foodchem.2018.04.029.[J].Food Chemistry, 2018, 261(2):51-56 [17]WU Q, CAI Q F, TAO Z P, et al.Purification and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from abalone (Haliotis discus hannai Ino) gonads[J].European Food Research & Technology, 2015, 240(1):137-145 [18]ZHANG J, XIN L, SHAN B, et al.PEAKS DB: de novo sequencing assisted database search for sensitive and accurate peptide identification[J].Molecule & Cell Proteomics, 2012, 11(4):M111- [19]谢雪琼, 钟婵, 孙乐常, 等.利用坛紫菜藻红蛋白制备脯氨酰内肽酶抑制肽[J].食品科学, 2019, 40(14):123-129 [20] LE MAUX S, NONGONINERMA A B, FITZGERALD R J.Peptide composition and dipeptidyl peptidase IV inhibitory properties of β-lactoglobulin hydrolysates having similar extents of hydrolysis while generated using different enzyme-to-substrate ratios[J]. Food Research International, 2017, 99: 84-90. DOI:10.1016/j.foodres.2017.05.012. [21]伍强, 蔡秋凤, 翁凌, 等.利用鲍鱼性腺制备抑制肽[J].中国食品学报, 2015, 15(10):97-105 [22] MINKIEWICZ P, IWANIAK A, DAREWICZ M.BIOPEP-UWM database of bioactive peptides: current opportunities[J]. International Journal of Molecular Sciences, 2019, 20(23). DOI:10.3390/ijms20235978. [23]LIU R, ZHOU L, ZHANG Y, et al.Rapid identification of dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides from Ruditapes philippinarum hydrolysate[J].Molecules, 2017, 22(10):1714-1725 [24]POWER O, NONGONIERMA A B, JAKEMAN P, et al.Food protein hydrolysates as a source of dipeptidyl peptidase IV inhibitory peptides for the management of type 2 diabetes[J].Proceedings of the Nutrition Society, 2014, 73(01):34-46 [25] NONGONINERMA A B, FITZGERALD R J.Features of dipeptidyl peptidase IV (DPP-IV) inhibitory peptides from dietary proteins[J]. Journal of Food Biochemistry, 2017: e12451. DOI:10.1111/jfbc.12451. [26]ZHANG Y, CHEN R, MA H, et al.Isolation and identification of dipeptidyl peptidase IV-inhibitory peptides from trypsinchymotrypsin-treated goat milk casein hydrolysates by 2D-TLC and LC-MSMS[J].Journal of Agriccultural and Food Chemistry, 2015, 63(40):8819-8828