【摘要】：【目的】敲除副產物途徑,提高重組大腸桿菌D-1,2,4-丁三醇(D-1,2,4-Butanetriol,BT)產量�！痉椒ā坷肦ed重組技術敲除木糖途徑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,考察其對重組菌生長、BT生產及副產物積累的影響。【結果】敲除xyl AB基因后,重組菌生物量降低57%,BT產量降低20%,單位菌體產量提高84%,木糖酸積累量提高52%。yag E或yjh H基因單獨缺失重組菌生物量分別提高10%和5%,BT產量提高36%和14%�；蚬餐笔Ш笾亟M菌生物量降低了21%,BT產量提高184%,達到2.44 g/L,單位菌體產量提高258%。而共同敲除兩途徑,生物量降低了72%,雖然單位菌體產量提高了約4倍,但BT產量僅提高43%。p H調控下,重組菌木糖酸積累量下降,BT產量進一步提高,最高達3.11 g/L�！窘Y論】xyl AB基因缺失后,雖有利于提高BT途徑的效率,但由于木糖無法進入PPP途徑及木糖酸積累,造成生物量降低,不利于BT合成。單獨敲除yag E或yjh H后BT產量略有提高,而共同敲除這兩基因更為有效地調整碳流向BT合成偏轉。兩途徑共同敲除利于BT的合成,但由于菌體量的減少,無法大量獲得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】： 江南大學糖化學與生物技術教育部重點實驗室;江南大學工業(yè)生物技術教育部重點實驗室工業(yè)微生物研究中心;江南大學化學與材料工程學院;
【基金】：江蘇省自然科學基金項目(No.BK20140138) 111項目(No.111-2-06) 江蘇省六大人才高峰(No.2014-XCL-017)~~
【分類號】：R378.2

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