固定化酶与游离酶相比,具有良好的稳定性和重复使用性,可用于酶抑制剂的快速筛选。该研究合成了一种新型的固定化α-葡萄糖苷酶3-MPBA/AG@aZIF-7/PDA,并对其进行了表征。此外,以Ile-Tyr-Gly(ITG,非竞争型抑制肽)与Cys-Tyr-Gly(CTG,竞争型抑制肽)为吸附模型,研究3-MPBA/AG@aZIF-7/PDA的吸附性能。结果显示,3-MPBA/AG@aZIF-7/PDA在优化的吸附条件下,对ITG和CTG的吸附量分别为191.06、129.50 mg/g。吸附动力学和吸附等温线研究表明,3-MPBA/AG@aZIF-7/PDA对ITG和CTG的吸附均符合拟一级吸附动力学模型,主要为物理吸附过程,吸附过程符合以单分子层吸附为主的Langmuir等温吸附模型,此外,研究发现该固定化酶能从复杂体系中有效分离出AG抑制肽。这些结果表明,固定化酶在分离纯化方面对功能肽的分离和富集具有重要的价值,而3-MPBA/AG@aZIF-7/PDA是一种很有潜力的高效分离α-葡萄糖苷酶抑制剂的材料。
Immobilized enzymes have good stability and reusability compared to free enzymes and can be used for rapid screening of enzyme inhibitors.In this study, a novel immobilized α-glucosidase 3-MPBA/AG@aZIF-7/PDA was synthesized and characterized.In addition, Ile-Tyr-Gly (ITG, non-competitive inhibitory peptide) and Cys-Tyr-Gly (CTG, competitive inhibitory peptide) were used as adsorption models to study the adsorption properties of 3-MPBA/AG@aZIF-7/PDA.Results showed that the adsorption capacity of 3-MPBA/AG@aZIF-7/PDA for ITG and CTG was 191.06 mg/g and 129.50 mg/g, respectively, under the optimized adsorption conditions.The adsorption kinetics and adsorption isotherm studies indicated that the adsorption of both ITG and CTG by 3-MPBA/AG@aZIF-7/PDA conformed to the proposed one-stage adsorption kinetic model, which was mainly a physical adsorption process, and the adsorption process conformed to the Langmuir isothermal adsorption model that was dominated by the adsorption of monomolecular layers.Moreover, it was found that the immobilized enzyme could efficiently isolate the AG inhibitory peptides from the complex system.These results indicate that the immobilized enzyme is of great value in isolation and enrichment of functional peptides, and 3-MPBA/AG@aZIF-7/PDA is a promising material for the efficient isolation of α-glucosidase inhibitors.
[1] WAN G Z, MA X H, JIN L, et al.α-glucosidase immobilization on magnetic core-shell metal-organic frameworks for inhibitor screening from traditional Chinese medicines[J].Colloids and Surfaces B:Biointerfaces, 2021, 205:111847.
[2] TAN K M, TESAR C, WILTON R, et al.Interaction of antidiabetic α-glucosidase inhibitors and gut bacteria α-glucosidase[J].Protein Science, 2018, 27(8):1498-1508.
[3] YIN Z H, ZHANG W, FENG F J, et al.α-Glucosidase inhibitors isolated from medicinal plants[J].Food Science and Human Wellness, 2014, 3(3-4):136-174.
[4] 刘平, 舒文娟, 冯秀, 等.阿卡波糖片治疗2型糖尿病的应用研究进展[J].中国处方药, 2020, 18(3):13-14.
LIU P, SHU W J, FENG X, et al.Research progress of acarbose tablets in the treatment of type 2 diabetes mellitus[J].Journal of China Prescription Drug, 2020, 18(3):13-14.
[5] 杨洁. 伏格列波糖片对2型糖尿病空腹血糖、餐后2 h血糖、糖化血红蛋白的影响[J].中国医药指南, 2019, 17(29):70-71.
YANG J.Effect of voglibose tablets on fasting blood glucose, 2 h postprandial blood glucose and glycosylated hemoglobin in type 2 diabetes mellitus[J].Guide of China Medicine, 2019, 17(29):70-71.
[6] WAN G Z, MA X H, JIN L, et al.Fabrication of a magnetic porous organic polymer for α-glucosidase immobilization and its application in inhibitor screening[J].Langmuir, 2023, 39(15):5239-5249.
[7] LEE H J, LEE H S, CHOI J W, et al.Novel tripeptides with α-glucosidase inhibitory activity isolated from silk cocoon hydrolysate[J].Journal of Agricultural and Food Chemistry, 2011, 59(21):11522-11525.
[8] ZHANG Y P, WU F H, HE Z P, et al.Optimization and molecular mechanism of novel α-glucosidase inhibitory peptides derived from Camellia seed cake through enzymatic hydrolysis[J].Foods, 2023, 12(2):393.
[9] YU Z P, YIN Y G, ZHAO W Z, et al.Novel peptides derived from egg white protein inhibiting alpha-glucosidase[J].Food Chemistry, 2011, 129(4):1376-1382.
[10] LI X H, SU Z R, LUO Y S, et al.Modelling of oligodextran production via an immobilized enzyme membrane reactor:Bioreaction-separation coupling mechanism[J].Separation and Purification Technology, 2022, 282:120024.
[11] MUJTABA N, JAHAN N, SULTANA B, et al.Isolation and characterization of antihypertensive peptides from soy bean protein[J].Brazilian Journal of Pharmaceutical Sciences, 2021, 57.DOI:10.1590/s2175-97902020000419061.
[12] LAN X D, LIAO D K, WU S G, et al.Rapid purification and characterization of angiotensin converting enzyme inhibitory peptides from lizard fish protein hydrolysates with magnetic affinity separation[J].Food Chemistry, 2015, 182:136-142.
[13] 孙美玲. 基于亲和色谱的皮氏蛾螺ACE抑制肽分离及其特性评价[D].大连:大连工业大学, 2017.
SUN M L.Isolation and characterization of ACE inhibitory peptides from Oncomelania pipiens based on affinity chromatography[D].Dalian:Dalian Polytechnic University, 2017.
[14] HIPPAUF F, HUETTNER C, LUNOW D, et al.Towards a continuous adsorption process for the enrichment of ACE-inhibiting peptides from food protein hydrolysates[J].Carbon, 2016, 107:116-123.
[15] LIU L, QIAO Z W, CUI X F, et al.Amino acid imprinted UiO-66 s for highly recognized adsorption of small angiotensin-converting-enzyme-inhibitory peptides[J].ACS Applied Materials & Interfaces, 2019, 11(26):23039-23049.
[16] HUETTNER C, HAGEMANN D, TROSCHKE E, et al.Tailoring the adsorption of ACE-inhibiting peptides by nitrogen functionalization of porous carbons[J].Langmuir, 2019, 35(30):9721-9731.
[17] 刘思媛, 刘精杏, 王则奋, 等.水相中GOx@ZIF-7/PDA的制备及酶学性能研究[J].食品与发酵工业, 2023, 49(18):157-164.
LIU S Y, LIU J X, WANG Z F, et al.Preparation of GOx@ZIF-7/PDA in aqueous phase and its enzymatic properties[J].Food and Fermentation Industries, 2023, 49(18):157-164.
[18] KHAN S N, SHAHEEN F, ALEEM U, et al.Peptide conjugates of 18β-glycyrrhetinic acid as potent inhibitors of α-glucosidase and AGEs-induced oxidation[J].European Journal of Pharmaceutical Sciences, 2022, 168:106045.
[19] 苏会东, 黄维华.D113大孔树脂吸附Ni2+的动力学与热力学研究[J].环境科学与技术, 2009, 32(10):49-52.
SU H D, HUANG W H.Kinetics and thermodynamics of sorption of Ni2+ macroporous adsorption resin D113[J].Environmental Science & Technology, 2009, 32(10):49-52.
[20] GEORGIANOS P, POURNARA A D, ANDREOU E K, et al.Composite materials based on a Zr4+ MOF and aluminosilicates for the simultaneous removal of cationic and anionic dyes from aqueous media[J].Molecules, 2023, 28(2):815.
[21] REN Y, LIANG K, JIN Y Q, et al.Identification and characterization of two novel α-glucosidase inhibitory oligopeptides from hemp (Cannabis sativa L.) seed protein[J].Journal of Functional Foods, 2016, 26:439-450.
[22] LIN Z X, RICHARDSON J J, ZHOU J J, et al.Direct synthesis of amorphous coordination polymers and metal-organic frameworks[J].Nature Reviews.Chemistry, 2023, 7(4):273-286.
[23] JIAO R, WANG Y M, PANG Y X, et al.Construction of macroporous β-glucosidase@MOFs by a metal competitive coordination and oxidation strategy for efficient cellulose conversion at 120 ℃[J].ACS Applied Materials & Interfaces, 2023, 15(6):8157-8168.
[24] MOHAMMAD M, RAZMJOU A, LIANG K, et al.Metal-organic-framework-based enzymatic microfluidic biosensor via surface patterning and biomineralization[J].ACS Applied Materials & Interfaces, 2019, 11(2):1807-1820.
[25] WANG Z F, ZHOU Q, LIU S Y, et al.Anchoring of polymer loops on enzyme-immobilized mesoporous ZIF-8 enhances the recognition selectivity of angiotensin-converting enzyme inhibitory peptides[J].Molecules, 2023, 28(7):3117.
[26] 肖志红, 吴红, 李昌珠, 等.固定化假丝酵母脂肪酶对光皮树油的吸附性能[J].中南林业科技大学学报, 2014, 34(2):117-121.
XIAO Z H, WU H, LI C Z, et al.Adsorption kinetics of immoblized Candida lipase to Swida wilsoniana oil[J].Journal of Central South University of Forestry & Technology, 2014, 34(2):117-121.
