化工学报
化工學報
화공학보
JOURNAL OF CHEMICAL INDUSY AND ENGINEERING (CHINA)
2015年
7期
2620-2627
,共8页
马鹏飞%蒙坚%周静%高海军
馬鵬飛%矇堅%週靜%高海軍
마붕비%몽견%주정%고해군
D-1,2,4-丁三醇%PTS系统%代谢%生物催化%合成生物学
D-1,2,4-丁三醇%PTS繫統%代謝%生物催化%閤成生物學
D-1,2,4-정삼순%PTS계통%대사%생물최화%합성생물학
D-1,2,4-butanetriol%PTS system%metabolism%biocatalysis%synthetic biology
1,2,4-丁三醇(1,2,4-butanetriol, BT)是一种重要的有机合成中间体。通过克隆表达恶臭假单胞菌(Pseudomonas putida ATCC12633)2-酮酸脱羧酶(mdlC)和新月柄杆菌(Caulobacter crescentus CB15)D-木糖脱氢酶(xdh),敲除木糖利用和 D-1,2,4-丁三醇合成中间代谢物分解途径中关键基因木糖异构酶(xylA)和2-酮酸醛缩酶(yjhH和yagE),重构大肠杆菌代谢网络,得到了能够将D-木糖转化为D-1,2,4-丁三醇的重组菌株。考察了温度、装液量、pH控制等条件对重组菌株合成D-1,2,4-丁三醇的影响,在适宜条件下发酵36 h后D-1,2,4-丁三醇产量达到3.96 g·L?1。探讨了葡萄糖利用与丁三醇合成的关系,通过敲除编码酶 IICBGlc的 ptsG 基因改造重组菌株的磷酸烯醇式丙酮酸葡萄糖转移酶(phosphoenolpyruvate:sugar phosphotransferase system, PTS)系统,菌株可以在利用葡萄糖生长的同时进行木糖的转化,具有更高的合成能力。
1,2,4-丁三醇(1,2,4-butanetriol, BT)是一種重要的有機閤成中間體。通過剋隆錶達噁臭假單胞菌(Pseudomonas putida ATCC12633)2-酮痠脫羧酶(mdlC)和新月柄桿菌(Caulobacter crescentus CB15)D-木糖脫氫酶(xdh),敲除木糖利用和 D-1,2,4-丁三醇閤成中間代謝物分解途徑中關鍵基因木糖異構酶(xylA)和2-酮痠醛縮酶(yjhH和yagE),重構大腸桿菌代謝網絡,得到瞭能夠將D-木糖轉化為D-1,2,4-丁三醇的重組菌株。攷察瞭溫度、裝液量、pH控製等條件對重組菌株閤成D-1,2,4-丁三醇的影響,在適宜條件下髮酵36 h後D-1,2,4-丁三醇產量達到3.96 g·L?1。探討瞭葡萄糖利用與丁三醇閤成的關繫,通過敲除編碼酶 IICBGlc的 ptsG 基因改造重組菌株的燐痠烯醇式丙酮痠葡萄糖轉移酶(phosphoenolpyruvate:sugar phosphotransferase system, PTS)繫統,菌株可以在利用葡萄糖生長的同時進行木糖的轉化,具有更高的閤成能力。
1,2,4-정삼순(1,2,4-butanetriol, BT)시일충중요적유궤합성중간체。통과극륭표체악취가단포균(Pseudomonas putida ATCC12633)2-동산탈최매(mdlC)화신월병간균(Caulobacter crescentus CB15)D-목당탈경매(xdh),고제목당이용화 D-1,2,4-정삼순합성중간대사물분해도경중관건기인목당이구매(xylA)화2-동산철축매(yjhH화yagE),중구대장간균대사망락,득도료능구장D-목당전화위D-1,2,4-정삼순적중조균주。고찰료온도、장액량、pH공제등조건대중조균주합성D-1,2,4-정삼순적영향,재괄의조건하발효36 h후D-1,2,4-정삼순산량체도3.96 g·L?1。탐토료포도당이용여정삼순합성적관계,통과고제편마매 IICBGlc적 ptsG 기인개조중조균주적린산희순식병동산포도당전이매(phosphoenolpyruvate:sugar phosphotransferase system, PTS)계통,균주가이재이용포도당생장적동시진행목당적전화,구유경고적합성능력。
1,2,4-Butanetriol (BT) is an important organic synthetic intermediate. In this study, the metabolic network of Escherichia coli was reconstructed by heterogeneously expressing a keto acid decarboxylase (mdlC) from Pseudomonas putida ATCC12633 and a D-xylose dehydrogenase (xdh) from Caulobacter crescentus CB15, and knocking out xylA, yjhH and yagE which were the genes of xylose utilization pathway and intermediary metabolite pathway for D-1,2,4-butanetriol synthesis. The recombinant strain could synthesize D-1,2,4-butanetriol directly using D-xylose as precursor. Culture conditions such as temperature, medium volume, pH of fermentation broth were investigated at the titer of D-1,2,4-butanetriol of 3.96 g·L?1 under suitable fermentation conditions. The relationship between glucose utilization and D-1,2,4-butanetriol synthesis was discussed. After modifying the phosphoenolpyruvate:sugar phosphotransferase system (PTS) by knocking out ptsG the reconstructed E.coli could utilize glucose and xylose simultaneously, leading to a higher D-1,2,4-butanetriol productivity.