本文選題：響應(yīng)面法 + 短小芽孢桿菌��； 參考：《應(yīng)用與環(huán)境生物學(xué)報》2016年03期

【摘要】：短小芽孢桿菌SCU11是由本實驗室分離的野生菌株BA06經(jīng)過復(fù)合誘變得到的高產(chǎn)堿性蛋白酶菌株,其產(chǎn)生的堿性蛋白酶在生物制革領(lǐng)域具有很好的應(yīng)用前景.為進一步提高菌株的蛋白酶產(chǎn)量以滿足工業(yè)生產(chǎn)的需求,本研究利用響應(yīng)面法優(yōu)化菌株產(chǎn)蛋白酶的發(fā)酵培養(yǎng)條件.在前期單因素實驗的基礎(chǔ)上,通過Plackett-Burman實驗設(shè)計、最陡爬坡實驗、中心組合實驗設(shè)計和響應(yīng)面分析法,確定了當黃豆粉濃度為53.3 g/L,溫度為28℃時,蛋白酶活有理論最大值8 884 U/m L,搖瓶實驗驗證酶活為8 768 U/m L,達到理論預(yù)測值的98.7%,比優(yōu)化前酶活提高了1倍.為了解優(yōu)化的發(fā)酵條件使蛋白酶產(chǎn)量提高的原因,對短小芽孢桿菌蛋白酶基因及相關(guān)調(diào)控基因的表達情況進行熒光定量PCR分析,結(jié)果顯示,優(yōu)化后5種蛋白酶基因的表達量上調(diào),1種表達量下調(diào);5種蛋白酶相關(guān)調(diào)控基因的表達量增加,2種基因表達量降低.推測是優(yōu)化后正調(diào)控基因表達量的增加以及負調(diào)控基因表達量的降低,促進了短小芽孢桿菌胞外蛋白酶基因轉(zhuǎn)錄水平的提高,導(dǎo)致胞外蛋白酶產(chǎn)量增加.本研究采用響應(yīng)面法優(yōu)化發(fā)酵條件使短小芽孢桿菌SCU11蛋白酶產(chǎn)量提高了1倍,并為闡明其表達調(diào)控機制奠定了基礎(chǔ).(圖3表7參24)
[Abstract]:Bacillus pumilus SCU11 is a high-yielding alkaline protease strain isolated from the wild strain BA06 in our laboratory. The alkaline protease produced by SCU11 has a good application prospect in the field of biological tannery. In order to further improve the protease production of the strain to meet the needs of industrial production, the response surface method was used to optimize the fermentation and culture conditions of the protease production of the strain. On the basis of the previous single-factor experiments, the Plackett-Burman experiment, the steepest climbing experiment, the central combination experiment design and the response surface analysis, the results show that when the concentration of soybean powder is 53.3 g / L and the temperature is 28 鈩，

本文編號：2112406