【摘要】：【目的】敲除副產(chǎn)物途徑,提高重組大腸桿菌D-1,2,4-丁三醇(D-1,2,4-Butanetriol,BT)產(chǎn)量�！痉椒ā坷肦ed重組技術(shù)敲除木糖途徑xyl AB基因及2-酮-3-脫氧木糖酸代謝途徑的yag E及yjh H基因,考察其對(duì)重組菌生長(zhǎng)、BT生產(chǎn)及副產(chǎn)物積累的影響�！窘Y(jié)果】敲除xyl AB基因后,重組菌生物量降低57%,BT產(chǎn)量降低20%,單位菌體產(chǎn)量提高84%,木糖酸積累量提高52%。yag E或yjh H基因單獨(dú)缺失重組菌生物量分別提高10%和5%,BT產(chǎn)量提高36%和14%。基因共同缺失后重組菌生物量降低了21%,BT產(chǎn)量提高184%,達(dá)到2.44 g/L,單位菌體產(chǎn)量提高258%。而共同敲除兩途徑,生物量降低了72%,雖然單位菌體產(chǎn)量提高了約4倍,但BT產(chǎn)量?jī)H提高43%。p H調(diào)控下,重組菌木糖酸積累量下降,BT產(chǎn)量進(jìn)一步提高,最高達(dá)3.11 g/L。【結(jié)論】xyl AB基因缺失后,雖有利于提高BT途徑的效率,但由于木糖無(wú)法進(jìn)入PPP途徑及木糖酸積累,造成生物量降低,不利于BT合成。單獨(dú)敲除yag E或yjh H后BT產(chǎn)量略有提高,而共同敲除這兩基因更為有效地調(diào)整碳流向BT合成偏轉(zhuǎn)。兩途徑共同敲除利于BT的合成,但由于菌體量的減少,無(wú)法大量獲得BT。
[Abstract]:[objective] to improve the production of recombinant Escherichia coli D-1O2C4- Butanetriol by knockout by-product pathway. [methods] the yag E and yjh H genes of xylose pathway xyl AB gene and 2-keto-3-deoxyxyxyxylic acid metabolic pathway were knocked out by Red recombinant technique. Effects of BT production and by-product accumulation. [results] after knockout of xyl AB gene, the biomass of recombinant bacteria decreased by 57% and BT yield decreased by 20%, and the yield per cell increased by 84%. The biomass of 52%.yag E or yjh H alone deleted recombinant bacteria increased by 10% and 5%, respectively, and the yield of BT increased by 36% and 14%, respectively. After gene deletion, the biomass of recombinant bacteria decreased by 21% and the yield of BT increased by 1844 g / L to 2.44 g / L, and the yield per unit cell increased by 258%. The biomass decreased by 72 times, but the yield of BT was increased only by 43.pH, and the accumulation of xylose acid was decreased, and the yield of BT was further increased. [conclusion] xyl AB gene deletion can improve the efficiency of BT pathway, but because xylose can not enter PPP pathway and xylose acid accumulation, the biomass is decreased, which is not conducive to BT synthesis. The yield of BT increased slightly after knockout of yag E or yjh H alone, and the co-knockout of these two genes was more effective in regulating the deflections of BT synthesis. The knockout of the two pathways is beneficial to the synthesis of BT, but BT. can not be obtained in large quantities because of the decrease of the number of bacteria.
【作者單位】： 江南大學(xué)糖化學(xué)與生物技術(shù)教育部重點(diǎn)實(shí)驗(yàn)室;江南大學(xué)工業(yè)生物技術(shù)教育部重點(diǎn)實(shí)驗(yàn)室工業(yè)微生物研究中心;江南大學(xué)化學(xué)與材料工程學(xué)院;
【基金】：江蘇省自然科學(xué)基金項(xiàng)目(No.BK20140138) 111項(xiàng)目(No.111-2-06) 江蘇省六大人才高峰(No.2014-XCL-017)~~
【分類號(hào)】：R378.2

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