为实现南美白对虾壳资源的绿色、高值化转化,利用碱性蛋白酶对虾壳中的蛋白质进行提取,水解产物与核糖进一步通过美拉德反应改性制备美拉德反应产物(Maillard reaction products, MRPs)。对MRPs进行红外表征,并研究其体外抗氧化活性和细胞内的抗氧化作用。MRPs制备条件为:核糖质量浓度0.025 g/mL、pH 5、反应温度90 ℃、反应时间8 h,此条件下的MRPs具有一定的还原能力,约为58 μmol TE/g,并且对ABTS阳离子自由基、DPPH自由基以及·OH清除率随浓度升高而增加,IC50值分别为(0.024±0.001)、(0.612±0.132)和(2.056 0±0.094)mg/mL;MRPs超滤获得的小分子质量和大分子质量组分均具有一定的抗氧化能力;此外,MRPs对人肾小管上皮细胞(HK-2)表现出良好的生物相容性,并且对H2O2诱导损伤的HK-2细胞具有显著的修复作用。因此,虾壳蛋白质水解物-核糖美拉德反应产物有望作为新型抗氧化剂,为虾壳废弃物资源的高值化利用提供新思路。
To realize the green and high-value utilization of protein resources in the shell of Litopenaeus vannamei, alkaline protease was used for hydrolyzing protein, and the hydrolysate was further modified with ribose by Maillard reaction to prepare Maillard reaction products (MRPs). MRPs were characterized by infrared spectroscopy and the antioxidant activities in vitro and intracellular were investigated. Results showed that the optimum conditions for preparing Maillard reaction products (MRPs) were obtained as follows, 2.5% of ribose concentration, 5.0 of initial pH value, 90 °C of temperature, and 8 h of time. Under these conditions, MRPs were an excellent antioxidant, exhibiting improving activity with increasing concentration, with a reducing capacity of about 58 μmol Trolox/g, and IC50 values of (0.024±0.001) mg/mL, (0.612±0.132) mg/mL, and (2.056 0±0.094) mg/mL for ABTS cationic radicals, DPPH free radicals, and ·OH radical, respectively. The low molecular weight fractions and high molecular weight fractions of MRPs obtained by ultrafiltration contributed to the antioxidant ability. In addition, MRPs showed good biocompatibility with human kidney cells (HK-2) and exhibited significant repair effects on the damage of HK-2 cells induced by H2O2. Therefore, shrimp shell protein hydrolysate-ribose MRPs are expected to be used as a novel antioxidant, which provides a new idea for the high-value utilization of shrimp shell waste resources.
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