摘要
利用超声波破碎法对Delftia sp. X-a12进行细胞破碎,研究超声波输出功率(W)、工作总时间(min)、菌体质量浓度(g/mL)对细胞破碎的效果,以香草酸脱羧酶酶比活为评判标准。通过单因素试验和响应面试验,获得香草酸脱羧酶酶比活较高的提取条件:超声波输出功率450 W、处理总时间18 min、菌体质量浓度0.12 g/mL,此条件下香草酸脱羧酶酶比活为541.27 U/mL。对粗酶液进行(NH4)2SO4分级沉淀,当(NH4)2SO4饱和度达到60%时,酶比活达到最大,为633.34 U/mL。
Cell disruption of Delftia sp. X-a12 was carried out by ultrasonication method. The ultrasonic output power, each irradiation time, total working time, and bacterial mass concentration were studied. The effect of cell disruption was judged by vanillic acid decarboxylase activity. Through single factor test and response surface test, the extraction condition of vanillic acid decarboxylase activity was obtained that ultrasonic output power 450 W, total treatment time 18 min, and cell mass concentration 0.12 g/mL. Under such condition, the vanillic acid decarboxylase enzyme ratio vitality was 541.27 U/mL. The crude enzyme solution was subjected to(NH_4)_2SO_4 fractionation precipitation. When the saturation of(NH_4)_2SO_4 reached60%, the enzyme activity reached the maximum, being 633.34 U/mL.
引文
[1]孙啸涛,王宗元,刘淼,等.涡旋辅助液液微萃取结合GC-MS法检测67种白酒中四甲基吡嗪、4-甲基愈创木酚和4-乙基愈创木酚[J].食品科学,2017,38(18):73-79.
[2]吴建峰.中国白酒中健康功能性成分四甲基吡嗪的研究[J].酿酒科技,2007(1):117-120.
[3]王少磊,曹荣升,沈芳,等.浓香型大曲中4-乙基愈创木酚产生菌的筛选及其鉴定[J].酿酒科技,2018(5):48-52.
[4]蔡瑞.脂环酸芽孢杆菌产愈创木酚代谢途径解析及检测控制方法研究[D].杨凌:西北农林科技大学,2016.
[5]景艳艳,陈茂彬,方尚玲,等.吡嗪类化合物在白云边酒中的变化[J].食品与营养科学,2017,6(4):215-222.
[6]王迪,曹方,栾静,等.超声波破碎法提取发酵性丝孢酵母胞内脂肪酶的条件优化[J].大连工业大学学报,2012,31(6):405-408.
[7]张荆,童群义.超声波破碎提取D-甘露糖异构酶条件的研究[J].食品工业科技,2009,30(10):247-249.
[8]张宽朝,全明吉.黑豆多酚氧化酶的分离纯化及其酶学特性研究[J].中国饲料,2012(19):38-41.
[9]刘亮,王如坤,程慧媛,等.水蜜桃多酚氧化酶分离纯化及其酶学特性研究[J].核农学报,2015,29(6):1121-1128.
[10]黄涛,杨洋,聂静然,等.桑叶多酚氧化酶分离及活性研究[J].食品科学,2008,29(12):473-476.
[11]尚宏丽,孟鑫,张挺.薏米蛋白提取及其SDS-PAGE电泳分析[J].中国农学通报,2012,28(18):260-265.
[12]曾广娟,李春敏,张新忠,等.适于SDS-PAGE分析的苹果叶片蛋白质提取方法[J].华北农学报,2009,24(2):75-78.
[13]杨玉红,杨建,张光宇,等.超声波破碎法提取羊肚菌发酵菌丝体生物活性肽条件的优化[J].食品科技,2014,39(7):252-257.
[14]闵钟熳,贾笑雨,解铁民,等.微生物发酵法提取米糠粕中可溶性膳食纤维的研究[J].中国酿造,2017,36(8):53-56.
[15]郭宇星,潘道东.超声波破碎法提取瑞士乳杆菌氨肽酶条件的优化[J].食品科学,2008(8):140-144.