本文選題：嗜熱果糖苷酶 + 基因克隆��； 參考：《中國食品添加劑》2017年04期

【摘要】：從嗜熱菌Thermotoga neapolitana DSM4359得到嗜熱果糖苷酶基因片段,以p ET-28a(+)為表達載體,在Escherichia.coil BL21(DE3)中高效表達。由SDS-PAGE檢測可得構建后嗜熱果糖苷酶的分子量約為60 KDa。通過單因素試驗和響應面法對該嗜熱果糖苷酶的產(chǎn)酶培養(yǎng)基進行優(yōu)化選擇,最終確定最佳產(chǎn)酶培養(yǎng)基為:蔗糖10.11 g/L,硝酸銨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。相比優(yōu)化前,經(jīng)優(yōu)化后的培養(yǎng)基所產(chǎn)嗜熱果糖苷酶酶活提高了近2.1倍。達到了提高嗜熱果糖苷酶表達量的目的。
[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.
【作者單位】： 吉林農(nóng)業(yè)大學食品科學與工程學院;
【基金】：吉林省教育廳“十二五”科學技術研究項目(吉教科合字[2015]第207號) 吉林省科技廳科技創(chuàng)新人才培育計劃項目(項目編號:20140519011JH)
【分類號】：Q55;Q78
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本文編號：1843899