PB试验和BBD响应面优化酶解鱼鳞制备抗菌肽工艺
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摘要
以酶解液的多肽含量和抑菌活性作为考察指标,采用酸性蛋白酶酶解制备草鱼鱼鳞抗菌肽。在单因素的基础上,运用PB试验设计进行影响因素的显著性筛选,确定出显著性影响因素为pH、温度和时间,进而通过BBD试验得到草鱼鱼鳞抗菌肽制备的最佳工艺条件为:pH 2.72、40.22℃、97.33min,在该条件下酶解液对金黄色葡萄球菌的抑菌圈直径为18.49mm,对大肠杆菌的抑菌圈直径为17.31 mm。结果表明,利用酸性蛋白酶酶解制备草鱼鱼鳞抗菌肽是行得通的,为草鱼鱼鳞等下脚料实现低值原料高值化提供了新的思路。
Taking the polypeptide content and bacteriostasis activity of enzymatic hydrolysate as the evaluation index, the antimicrobial peptide of grass carp scale was prepared by enzymatic hydrolysis with acid protease. On the basis of single factor, the Plackett-Burman test design was used to analyze the significance of influencing factors. The influencing factors were determined as enzymatic pH, enzymatic temperature and enzymatic hydrolysis time, and then the antimicrobial peptide of grass carp scale was obtained by Box-Behnken test. The optimum technological conditions were as follows: enzymatic hydrolysis pH 2.72, enzymatic hydrolysis temperature 40.22 C and enzymatic hydrolysis time 97.33 min. Under these conditions, the diameter of the inhibitory zone of the enzymatic hydrolysate against Staphylococcus aureus was 18.49 mm, and the diameter of the inhibitory zone against Escherichia coli was 17.31 mm. The results showed that it was feasible to prepare antimicrobial peptide from grass carp scale by enzymatic hydrolysis of acid protease, which provided a new idea for realizing high value of low value raw materials for grass carp scale and other waste materials.
Taking the polypeptide content and bacteriostasis activity of enzymatic hydrolysate as the evaluation index, the antimicrobial peptide of grass carp scale was prepared by enzymatic hydrolysis with acid protease. On the basis of single factor, the Plackett-Burman test design was used to analyze the significance of influencing factors. The influencing factors were determined as enzymatic pH, enzymatic temperature and enzymatic hydrolysis time, and then the antimicrobial peptide of grass carp scale was obtained by Box-Behnken test. The optimum technological conditions were as follows: enzymatic hydrolysis pH 2.72, enzymatic hydrolysis temperature 40.22 C and enzymatic hydrolysis time 97.33 min. Under these conditions, the diameter of the inhibitory zone of the enzymatic hydrolysate against Staphylococcus aureus was 18.49 mm, and the diameter of the inhibitory zone against Escherichia coli was 17.31 mm. The results showed that it was feasible to prepare antimicrobial peptide from grass carp scale by enzymatic hydrolysis of acid protease, which provided a new idea for realizing high value of low value raw materials for grass carp scale and other waste materials.
引文
[1]李燕,王嘉榕,孙红宾.抗菌肽的制备和抗菌机制研究进展[J].生物医学工程学杂志,2015,32(2):465-469.
[2]蒋妍,熊佳,余训培,等.草鱼鱼鳞蛋白酶解物对鲢鱼鱼丸贮藏及品质特性的影响[J].中国农业大学学报,2013,18(6).
[3]金文刚,吴海涛,朱蓓薇,等.虾夷扇贝生殖腺多肽的制备及分离[J].大连工业大学学报,2011,30(6):391-395.
[4]刘如运.几种常用抑菌试验方法的评价及比较[J].现代企业教育,2013,No.433(14):341-342.
[2]蒋妍,熊佳,余训培,等.草鱼鱼鳞蛋白酶解物对鲢鱼鱼丸贮藏及品质特性的影响[J].中国农业大学学报,2013,18(6).
[3]金文刚,吴海涛,朱蓓薇,等.虾夷扇贝生殖腺多肽的制备及分离[J].大连工业大学学报,2011,30(6):391-395.
[4]刘如运.几种常用抑菌试验方法的评价及比较[J].现代企业教育,2013,No.433(14):341-342.