發(fā)布時(shí)間：2018-06-11 11:14

  本文選題：間苯三酚 + MarA　； 參考：《生物技術(shù)通報(bào)》2017年12期

【摘要】：為進(jìn)一步提高生物法合成間苯三酚的效率,探究生物合成間苯三酚的分子機(jī)理以及更加高效的發(fā)酵條件和培養(yǎng)基成分。對(duì)來(lái)自熒光假單胞菌(Pseudomonas fluorescens)的phl D基因進(jìn)行了克隆,在BL21(DE3)中重建間苯三酚的胞內(nèi)合成途徑,超表達(dá)多重抗藥基因正向調(diào)節(jié)因子(mar A)和乙酰輔酶A羧化酶(ACCase),并分析不同碳源及培養(yǎng)基離子環(huán)境對(duì)間苯三酚積累量的影響。結(jié)果顯示,成功構(gòu)建間苯三酚胞內(nèi)合成途徑,產(chǎn)量達(dá)到450 mg/L,mar A基因能夠提高工程菌對(duì)間苯三酚抵抗力,使間苯三酚產(chǎn)量提高到1 060 mg/L;ACCase能夠提高胞內(nèi)丙二酰輔酶A的含量,使間苯三酚產(chǎn)量提高到2 120 mg/L;葡萄糖、甘油和丙酮酸鈉三種碳源的比較中,葡萄糖對(duì)合成間苯三酚最為有利,分別是甘油和丙酮酸鈉的1.8倍和2.4倍;培養(yǎng)基中鈣鎂離子對(duì)間苯三酚的合成沒有影響。mar A和ACCase基因的超表達(dá)能大大提高間苯三酚生物合成的效率,以葡萄糖為碳源的無(wú)鈣鎂培養(yǎng)基為間苯三酚發(fā)酵的最適培養(yǎng)基。
[Abstract]:In order to further improve the efficiency of biosynthesis of benzol three, the molecular mechanism of biosynthesis of benzol and the more efficient fermentation conditions and medium components were explored. The Phl D gene from Pseudomonas fluorescens (Pseudomonas fluorescens) was cloned and the intracellular synthesis pathway was reconstructed in BL21 (DE3), and the overexpression was overexpressed in BL21 (DE3). The effect of multiple anti drug gene positive regulator (Mar A) and acetyl coenzyme A carboxylase (ACCase), and the effects of different carbon sources and medium ion environment on the accumulation of benzol three were analyzed. The results showed that the intracellular synthesis pathway of phenylephol was successfully constructed, the yield reached 450 mg/L, and mar A gene could improve the resistance of engineering bacteria to the phenylephenol three. The yield of three phenol is increased to 1060 mg/L; ACCase can increase the content of intracellular A two acyl coenzyme and increase the yield of interphenolol to 2120 mg/L; in the comparison of three carbon sources of glucose, glycerin and sodium pyruvate, glucose is most favorable to the synthesis of interphenols, 1.8 times and 2.4 times of glycerol and sodium propionate, respectively. The synthesis of benzol three has no effect on the overexpression of.Mar A and ACCase gene, which can greatly improve the efficiency of biosynthesis of phenolol. The non calcium magnesium medium with glucose as the carbon source is the most suitable medium for the fermentation of phenylene three.
【作者單位】： 濰坊科技學(xué)院化工與環(huán)境學(xué)院;壽光富康制藥研發(fā)部;
【基金】：海洋生物醫(yī)藥間苯三酚等制劑國(guó)際化發(fā)展能力建設(shè)項(xiàng)目[魯財(cái)建指2014(No.18)]
【分類號(hào)】：TQ243.12

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