响应面法优化南极磷虾亚铁螯合肽制备工艺及其理化性质
Optimization of Preparation of Iron-chelating Peptides from Antarctic Krill by Response Surface Methodology and Its Physicochemical Properties
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摘要: 以南极磷虾为原料,酶解制备亚铁螯合肽。以亚铁螯合率为主要指标,水解度为辅助指标,在单因素实验基础上,利用响应面优化制备南极磷虾亚铁螯合肽(Antarctic krill iron-chelating peptides,AKIP),并对其理化性质进行分析。结果表明:Alcalase 2.4 L是南极磷虾酶解的最适用酶,最佳酶解条件为:酶解温度57.6 ℃,加酶量5.5%,pH8.8,酶解时间5 h,在此条件下酶解液的亚铁螯合率为77.77%±0.72%,水解度为23.22%±0.16%,实测结果与预测值符合良好。氨基酸组成分析表明,南极磷虾亚铁螯合肽具有良好的亚铁螯合活性位点。AKIP的相对分子质量主要分布在180~1000 Da之间,约占89.28%。体外抗氧化实验表明,AKIP对DPPH自由基和羟自由基均有良好的清除能力(IC50分别为1.92、2.09 mg/mL),且呈现较好的量效关系,在食源性螯合肽和天然抗氧化剂领域具有一定的开发利用价值。Abstract: Iron-chelating peptide was prepared from Antarctic krill by enzymatic hydrolysis. With iron-chelating rate as the main indicator,and the degree of hydrolysis(DH)as indicator,the hydrolysis process was optimized using one-factor-at-a-time method and response surface methodology(RSM). The physicochemical properties of hydrolysates obtained under the optimized conditions were analyzed. Results showed that Alcalase 2.4 L was the suitable enzyme. The optimal hydrolysis conditions were as follows:Hydrolysis temperature 57.6 ℃,enzyme/substrate ratio 5.5%,pH8.8,hydrolysis time 5 h. The actual iron-chelating rate of the peptides prepared under the(optimized condition was 77.77%±0.72%,DH was 23.22%±0.16%,which was in good agreement with the predicted value. Amino acid composition analysis showed that the Antarctic krill iron-chelating peptides(AKIP)had good iron-chelating sites. The relative molecular weight of AKIP was amount of 180~1000 Da and accounted for 89.28%. In vitro antioxidant experiments showed that,AKIP had good scavenging capacity for DPPH free radicals and hydroxyl free radicals(IC50 was 1.92 and 2.09 mg/mL,respectively),and showed a good dose-effect relationship. This study AKIP had a certain value in food-protein derived chelating peptides and natural antioxidants fields.