食品科学2018，Vol.39Issue(6)：155-161,7.DOI:10.7506/spkx1002-6630-201806025
响应面试验优化双酶酶解法制备鱼鳞抗菌肽工艺及其抑菌性能分析
Optimization of Preparation of Antimicrobial Peptides by Two-Step Enzymatic Hydrolysis of Fish Scales Using Response Surface Methodology and Antimicrobial Activity of Purified Antimicrobial Peptide
摘要
Abstract
This study aimed to prepare antimicrobial peptides (AMPs) by two-step enzymatic hydrolysis of crucian carp scales and to evaluate the antimicrobial activity of purified AMPs.Hydrolysis conditions were optimized by one-factor-at-a-time method and response surface methodology based on the diameter of inhibition zones against tested bacteria.The enzymatic hydrolysate prepared using optimized conditions was then purified by Sephadex G-25 column chromatography,yielding only a single peak (G2) with antimicrobial activity.The minimum inhibitory concentration (MIC) of G2 was tested.The results indicated that a solid-to-liquid ratio of 30 g/100 mL and sequential hydrolysis with alcalase at pH 9.5 and 55 ℃ for 62 min followed by acid protease at pH 3.0 and 34.4 ℃ for another 3 h were found to be the optimal conditions to obtain a greater diameter of inhibition zone against Hbrio parahemolyticus of 27.72 mm,which was well matched with the predicted value (27.37 mm).The MIC of G2 was 1.56 μg/mL against Pseudomonas and Shewanella putrefaciens,and 6.25 μg/mL against Escherichia coli,Staphylococcus aureus,Salmonella choleraescens,V.parahemolyticus and Bacillus subtilis.In conclusion,AMPs derived from freshwater fish scales by stepwise enzymatic hydrolysis possess strong antimicrobial activity.关键词
酶解/鱼鳞/抗菌肽/响应面法/抑菌性Key words
enzymatic hydrolysis/fish scales/antimicrobial peptides/response surface methodology/antimicrobial activity分类
轻工纺织引用本文复制引用
施永清,王巧巧,吴丹丽,李晓玉,蔡路昀,励建荣..响应面试验优化双酶酶解法制备鱼鳞抗菌肽工艺及其抑菌性能分析[J].食品科学,2018,39(6):155-161,7.基金项目
食品科学与工程浙江省重中之重一级学科资助项目(2017SIAR221) （2017SIAR221）
浙江工商大学大学生科研创新基金项目(CX201710009) （CX201710009）
浙江工商大学2015年食品学院卓越食品人才(学生)科技培育项目(20151214) （学生）
辽宁省科技厅农业攻关及成果产业化项目(2015103020) （2015103020）
