嗜热果糖苷酶的基因克隆及表达优化

发布时间：2018-05-04 17:20

  本文选题：嗜热果糖苷酶 + 基因克隆　； 参考：《中国食品添加剂》2017年04期

【摘要】：从嗜热菌Thermotoga neapolitana DSM4359得到嗜热果糖苷酶基因片段,以p ET-28a(+)为表达载体,在Escherichia.coil BL21(DE3)中高效表达。由SDS-PAGE检测可得构建后嗜热果糖苷酶的分子量约为60 KDa。通过单因素试验和响应面法对该嗜热果糖苷酶的产酶培养基进行优化选择,最终确定最佳产酶培养基为:蔗糖10.11 g/L,硝酸铵12.30 g/L,安琪酵母浸粉FM905 8.24 g/L,MgSO_4·7H_2O 5.61 mmol/L,NaCl 10 g/L。相比优化前,经优化后的培养基所产嗜热果糖苷酶酶活提高了近2.1倍。达到了提高嗜热果糖苷酶表达量的目的。
[Abstract]:The thermophilic fructosidase gene fragment was obtained from thermophilic bacteria Thermotoga neapolitana DSM4359 and was highly expressed in Escherichia.coil BL21DE3 using pET-28a () as expression vector. The molecular weight of the constructed thermophilic glucosidase was about 60 KDa. detected by SDS-PAGE. Single factor test and response surface method were used to optimize the enzyme production medium of the thermophilic fructosidase. The optimum medium was determined as follows: sucrose 10.11 g / L, ammonium nitrate 12.30 g / L, An Qi yeast extract FM905 8.24 g / L, MgSO4 7H_2O 5.61 mmol / L NaCl 10 g / L. Compared with that before optimization, the thermotropic fructosidase activity of the optimized medium was increased by nearly 2.1 times. The purpose of increasing the expression of thermophilic fructosidase was achieved.
【作者单位】： 吉林农业大学食品科学与工程学院;
【基金】：吉林省教育厅“十二五”科学技术研究项目(吉教科合字[2015]第207号) 吉林省科技厅科技创新人才培育计划项目(项目编号:20140519011JH)
【分类号】：Q55;Q78
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本文编号：1843899