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当前位置: 首页 > 文章 > 一种天然材料复合体系固定化酒用双功能酶的研究 食品工业科技 2014,35 (14) 186-191+196
Position: Home > Articles > Study on an immobilized bifunctional enzyme applied in rice wine with natural materials Science and Technology of Food Industry 2014,35 (14) 186-191+196

一种天然材料复合体系固定化酒用双功能酶的研究

作  者：
查小红;杨广明;田亚平
单  位：
江南大学工业生物技术教育部重点实验室
关键词：
固定化;双功能酶;黄酒;氨基甲酸乙酯
摘  要：
运用吸附包埋交联的复合方法固定化普罗威登斯菌Providencia sp.JNB815所产的具有降解尿素和氨基甲酸乙酯活性的双功能酶。以尿素降解酶活性为指标,单因素优化固定化条件为:壳聚糖和明胶浓度分别为4.0%、2.0%,京尼平浓度0.4%,30℃恒温水浴振荡6h进行交联,交联后的微球固定化酶的时间为8h。固定化酶催化氨基甲酸乙酯(EC)和尿素反应时,最适pH分别为4.5和4.0,最适温度分别为60、40℃。固定化EC降解酶酶活在温度20~60℃,pH5.0~6.5之间相对稳定;固定化脲酶酶活在温度20~50℃,pH4.0~6.5之间相对稳定。固定化酶重复使用10次后,对尿素和EC的酶活残留率分别为80%和30%。在32℃,摇床转速100r/min的条件下,固定化尿素降解酶加酶量为0.04U/mL黄酒时,固定化酶处理黄酒20h后,尿素去除率可达93.03%;同样条件下,黄酒中固定化EC降解酶加酶量为0.17U/mL时,处理黄酒20h后,EC去除率可达56.60%。GC-MS结果显示固定化酶处理后的黄酒风味物质变化不大。
译  名：
Study on an immobilized bifunctional enzyme applied in rice wine with natural materials
作  者：
ZHA Xiao-hong;YANG Guang-ming;TIAN Ya-ping;Key Laboratory of Industrial Biotechnology,Ministry of Education Jiangnan University;
关键词：
immobilization;;bifunctional enzyme;;wine;;urethane
摘  要：
By using a compound method of adsorption,embedment and crosslink,a bifunctional enzyme produced by Providencia sp. JNB815 with both urease and urethanase activity was immobilized. With urea degrading activity as an indicator,the optimum immobilization conditions through single factor experiments were as follows:chitosan 4.0%,gelatin 2.0%,0.4% of genipin and the crosslink was conducted at 30℃ with concussion for 6h. Then the crosslinked microspheres were used to immobilize the enzyme for 8h. The optimum pH was 4.5 and 4.0,and the optimum temperature was 60℃ and 40℃,respectly when immobilized enzyme catalyzed urethane and urea. The immobilized enzyme activity to EC maintained stability when the temperature was in the range of 20~60℃ and pH 5.0 to 6.5. The immobilized enzyme activity to urea maintained stability when the temperature was in the range of 20~50℃ and pH 4.0 to 6.5.It retained 80% and 30% of the original catalytic avtivity to urea and EC,respectively after being used 10 times. Under the condition of 32℃ and100r/min,0.04U urease was added to per mL rice wine. After being processed for 20h,the removing rate of urea in wine reached 93.03%. Under the same condition,the removing rate of EC in wine was 56.60% after20h,when the amount of urethanase added was 0.17U per mL wine. In addition,the treatment with immobilized enzyme showed neglectable effect on the detected flavor substances in the wine.

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