红毛藻抗氧化肽的菠萝蛋白酶酶解制备工艺优化及其活性评价

Optimization of Preparation of Antioxidant Peptides from Bangia fuscopurpurea (Dillw.) Lyngb. Enzymolysised by Bromelain and Evaluation of Its Activity

以红毛藻(Bangia fuscopurpurea (Dillw.) Lyngb.)蛋白为原料,经菠萝蛋白酶水解制备抗氧化肽,为红毛藻的深加工提供理论依据.以酶解液对DPPH·的?清除率为响应值,考察酶解温度、酶解时间、酶解pH、酶底比对酶解液抗氧化活性的影响.在单因素试验基础上,通过L_9(3~4)正交试验优化红毛藻抗氧化肽的酶解制备工艺,并以酶解产物对O_2~ˉ·的清除能力和还原力为指标评价酶解产物的抗氧化活性.结果表明,用菠萝蛋白酶酶解红毛藻蛋白制备抗氧化肽的最优工艺参数为:酶解温度45℃、酶解时间90 min、酶解pH7.5、酶底比1200U/g.在最优酶解工艺条件下,红毛藻蛋白的水解度达到(10.74±0.13)%,酶解液中多肽浓度为(6.98±0.07) mg/mL,酶解产物对DPPH·和O_2~ˉ·的半数清除浓度(IC_(50))值分别为(0.4403±0.0142) mg/mL和(4.2600±0.0180) mg/mL,且具备一定的还原力.

Antioxidant peptides were prepared from the proteins of Bangia fuscopurpurea(Dillw.)Lyngb.enzymolysised by bromelain to provide a theoretical basis for the deep processing of B.fuscopurpurea.Using the scavenging rate of 1,1-Diphenyl-2-picrylhydrazyl radical(DPPH·)by the enzymatic hydrolysates as response value,the effects of single-factors including enzymolysis temperature,time,and pH and enzyme-to-substrate ratio([E][/S])on the antioxidant activity of the enzymatic hydrolysates were investigated.On the basis of a single factor experiment,L9(34)orthogonal experiment was used to optimize the B.fuscopurpurea antioxidant peptide enzyme preparation technology.This was accomplished by enzymolysis of hydrolysates to(O2ˉ·)removal ability and reducing power as an index and evaluation of the antioxidant activity of enzyme hydrolysates.The optimal parameters for hydrolyzing the proteins of B.fuscopurpurea to obtain antioxidant peptides by bromelain were found to be as follows:enzymolysis temperature 45℃,time 90 min,ands pH7.5 and[E][/S]1200 U/g.Under the optimal conditions,the degree of hydrolysis of B.fuscopurpurea protein reached(10.74±0.13)%,and the concentration of peptides was(6.98±0.07)mg/mL.The half-scavenging concentration(IC50)values of the enzymatic hydrolysates against DPPH·and O2ˉ·were(0.4403±0.0142)mg/mL and(4.2600±0.0180)mg/mL respectively,and the enzymatic hydrolysates had a certain degree of reducing power.