超声破碎辅助蜗牛酶提取杏鲍菇蛋白工艺优化

doi:10.7506/spkx1002-6630-201622012

摘要/Abstract

摘要： 为显著提高杏鲍菇子实体蛋白提取率，采用超声波破碎辅助蜗牛酶水解充分破碎菌体细胞壁。综合应用响应面、正交设计与人工神经网络模型相结合的试验设计方法，分别对超声破碎处理和蜗牛酶水解条件进行了优化，同时与直接热水碱提蛋白法进行了比较。结果表明，超声处理条件为超声功率300 W、超声时间26 min、水料比1.95∶5（mL/g），在此条件下破碎效果最好，提取率达到了48.82%；最佳蜗牛酶水解条件为温度42.8 ℃、pH 4.8、酶用量7%、时间1.5 h，在该条件下蛋白提取率为75.92%，相较热水碱提法提高了33.35%，结果表明利用超声辅助蜗牛酶水解杏鲍菇细胞壁能显著提高蛋白提取率。

关键词: 杏鲍菇, 蛋白, 超声破碎法, 蜗牛酶, 人工神经网络模型

Abstract: The aim of this work was to enhance the efficiency of protein extraction from fruit bodies of Pleurotus eryngii by cell disruption using ultrasonic irradiation followed by snailase hydrolysis. Response surface methodology, orthogonal array design and artificial neural network model were used in combination to optimize the conditions for ultrasonic irradiation and subsequent enzymatic hydrolysis, respectively. The protein extraction efficiencies of the presented method and the hot alkali method were compared. The best efficiency of cell disruption was achieved upon ultrasonic treatment for 26 min at 300 W power with a solid-to-water ratio of 1.95:5 (mL/g), which resulted in a protein yield of 48.82%. The optimum hydrolysis conditions that provided maximum protein yield of 75.92% were determined as 42.8 ℃, 4.8, 7% and 1.5 h for temperature, pH, enzyme dosage and hydrolysis time, respectively, which was increased by 33.35% compared with hot alkali extraction. These results showed that ultrasonic-assisted snailase hydrolysis could significantly increase the efficiency of protein extraction from P. eryngii fruit bodies.

Key words: Pleurotus eryngii, protein, ultrasonic-assisted extraction, snailase, artificial neural network

中图分类号:

TS255.2

漆倩涯,贠建民*,黄玉琴,白杰,张紊玮,艾对元. 超声破碎辅助蜗牛酶提取杏鲍菇蛋白工艺优化[J]. 食品科学, 2016, 37(22): 85-91.

QI Qianya, YUN Jianmin*, HUANG Yuqin, BAI Jie, ZHANG Wenwei, AI Duiyuan. Optimization of Ultrasonic-Assisted Snailase Hydrolysis for Extraction of Protein from Pleurotus eryngii[J]. FOOD SCIENCE, 2016, 37(22): 85-91.

参考文献

[1] 刁治民, 杨秀玲, 韩彦艳, 等. 蕈菌工程学[M]. 杨凌: 西北农林科技大学出版社, 2011.
[2] 王凤芳. 杏鲍菇中营养成分的分析测定[J]. 食品科学, 2002, 23(4): 132-135. DOI:10.3321/j.issn:1002-6630.2002.04.035.
[3] 宋爱荣, 岳运勇, 徐坤, 等. 4 个杏鲍菇品种的氨基酸分析与比较[J]. 菌物研究, 2005, 3(4): 11-14. DOI:10.3969/j.issn.1672-3538.2005.04.003.
[4] 李昊剑, 杨良嵘, 魏雪团, 等. 功能性短肽的分类及其酶解制备方法[J]. 食品工业科技, 2012, 33(17): 373-377. DOI:10.13386/j.issn1002-0306.2012.17.054.
[5] 励建荣, 封平. 功能肽的研究进展[J]. 食品科学, 2004, 25(11): 415-419. DOI:10.3321/j.issn:1002-6630.2004.11.110.
[6] 王成, 冯婷, 薛晓珍, 等. 新疆阿魏菇与杏鲍菇中氨基酸含量的分析研究[J]. 氨基酸和生物资源, 2013, 35(2): 56-58. DOI:10.14188/j.ajsh.2013.02.011.
[7] KO S C, KANG M C, LEE J K, et al. Effect of angiotensin I-converting enzyme (ACE) inhibitory peptide purified from enzymatic hydrolysates of Styela plicata[J]. European Food Research and Technology, 2011, 233(6): 915-922. DOI:10.1007/s00217-011-1585-7.
[8] 张梦甜, 杨文建, 裴斐, 等. 响应面法优化酶法制备杏鲍菇蛋白及其营养评价[J]. 食品科学, 2015, 36(13): 125-130. DOI:10.7506/spkx1002-6630-201513024.
[9] 马泽青, 张宝善, 赵舒欣, 等. 响应面法超声波辅助提取双孢蘑菇蛋白工艺优化[J]. 食品工业科技, 2012, 33(23): 229-233. DOI:10.13386/j.issn1002-0306.2012.23.044.
[10] 陈建军. 基于MEMS技术的细胞破碎微流控芯片系统研究[D]. 武汉: 华中科技大学, 2010.
[11] HJORTKJAER J. Rhodium complex catalyzed hydroformylation. The relation between effects of CO pressure and phosphine concentration[J]. Journal of Molecular Catalysis, 1979, 5(5): 377-384. DOI:10.1016/0304-5102(79)80013-9.
[12] GESTEIRA T F, COULSOM-THOMAS V J, OGATA F T, et al. A novel approach for the characterisation of proteoglycans and biosynthetic enzymes in a snail model[J]. Biochimicaet Biophysica Acta (BBA)-Proteins and Proteomics, 2011, 1814(12): 1862-1869. DOI:10.1007/s10719-011-9334-5.
[13] 陆晨, 邹雨虹, 张士康, 等. 响应面法优化超声辅助提取茶渣蛋白质的工艺条件[J]. 食品与生物技术学报, 2012, 31(3): 319-325.
[14] 张涛, 吴昌英, 张永模, 等. 蜗牛酶降解壳聚糖制备壳寡糖的研究[J]. 华西药学杂志, 2010, 25(3): 313-315. DOI:10.13375/j.cnki.wcjps.2010.03.053.
[15] 席甜, 冯翠萍, 程菲儿, 等. 杏鲍菇多肽的制备[J]. 山西农业大学学报(自然科学版), 2015, 35(1): 102-107. DOI:10.13842/j.cnki.issn1671-8151.2015.01.019.
[16] BRADFORD M. A rapid and sensitive method for the quantitation of protein using the principle of ptotein-dye binding[J]. Analytical Biochemistry, 1976, 72(1/2): 248-254. DOI:10.1016/0003-2697(76)90527-3.
[17] 桂现才. BP神经网络在MATLAB上的实现与应用[J]. 湛江师范学院学报, 2004, 25(3): 79-83. DOI:10.3969/j.issn.1006-4702.2004.03.021.
[18] 闵惜琳, 刘国华. 用MATLAB神经网络工具箱开发BP网络应用[J]. 计算机应用, 2001, 21(8): 163-164.
[19] 宋勇强, 贠建民, 安志刚, 等. 基于正交设计与人工神经网络模型的醋酸菌A3菌株醋酸发酵条件优化[J]. 食品工业科技, 2013, 34(5): 142-146. DOI:10.13386/j.issn1002-0306.2013.05.059.
[20] CHEN C R, RAMASWAMY H S, PRASHER S O. Dynamic modeling of retort processing using neural networks[J]. Journal of Food Processing and Preservation, 2002, 26(2): 91-112. DOI:10.1111/j.1745-4549.2002.tb00855.x.
[21] 陶然, 何东平, 胡传荣, 等. 超声波辅助提取芝麻蛋白的工艺研究[J]. 中国油脂, 2014, 39(5): 23-26.
[22] 许凤, 王长远. 响应面法优化物理辅助碱法提取米糠蛋白工艺[J]. 食品科学, 2014, 35(20): 11-16. DOI:10.7506/spkx1002-6630-201420003.
[23] 郭宇星. 瑞士乳杆菌蛋白酶的分离纯化及其水解乳蛋白产生ACE抑制肽研究[D]. 南京: 南京师范大学, 2009.
[24] LIU C L, LIN T H, JUANG R S. Optimization of recombinant hexaoligochitin—producing chitinaseproduction with response surface methodology[J]. Carbohydrate Polymers, 2013, 62(10): 518-522.
[25] GHORBEL B O, HAJJI S, YUNES I, et al. Optimization of chitin extraction from shrimp waste with Bacillus pumilus A1 using response surface metho dology[J]. International Journal of Biological Macromolecules, 2013, 61(2): 243-250. DOI:10.1016/j.ijbiomac.2013.07.001.
[26] 张晶. 人工神经网络在响应曲面法实验优化设计中的应用研究[D]. 西宁: 青海师范大学, 2014.