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不同葡萄糖轉(zhuǎn)運(yùn)途徑相關(guān)基因的過(guò)表達(dá)對(duì)菌株葡萄糖代謝及產(chǎn)物合成的影響

發(fā)布時(shí)間:2019-01-19 18:35
【摘要】:磷酸烯醇式丙酮酸-葡萄糖磷酸轉(zhuǎn)移酶系統(tǒng)(phosphoenolpyruvate-dependent glucose phosphotransferase system,PTS~(Glc))和不依賴于PTS~(Glc)系統(tǒng)葡萄糖轉(zhuǎn)運(yùn)系統(tǒng)在葡萄糖轉(zhuǎn)運(yùn)和磷酸化過(guò)程中起重要作用。該文通過(guò)在葡萄糖代謝緩慢的L-賴氨酸產(chǎn)生菌Corynebacterium glutamicum(C.glutamicum)ZL-8中過(guò)表達(dá)肌醇透性酶基因(iol T1)、葡萄糖激酶基因(ppgk)和EIIGlc基因(pts G),研究葡萄糖轉(zhuǎn)運(yùn)途徑相關(guān)基因?qū)ζ咸烟谴x速率、L-賴氨酸及副產(chǎn)物合成的影響。與出發(fā)菌株C.glutamicum ZL-8相比,重組菌C.glutamicum ZL-8/p ECXK-99E-ppgkiol T1(葡萄糖激酶酶活力提高了768%)和C.glutamicum ZL-8/p DXW-8-pts G(PTS~(Glc)系統(tǒng)酶活力提高了62.7%)葡萄糖消耗速率分別提高了16.7%和27.3%,L-賴氨酸產(chǎn)量分別提高了24.7%和16.4%。此外,有機(jī)酸和氨基酸分析結(jié)果表明,過(guò)表達(dá)不同葡萄糖轉(zhuǎn)運(yùn)途徑基因?qū)τ袡C(jī)酸(丙酮酸、乳酸和乙酸)和副產(chǎn)物氨基酸(丙氨酸、纈氨酸、天冬氨酸)具有不同的影響。
[Abstract]:Phosphoenolpyruvate-glucose phosphotransferase system (phosphoenolpyruvate-dependent glucose phosphotransferase system,PTS~ (Glc) and PTS~ (Glc) independent glucose transport system) plays an important role in the process of glucose transport and phosphorylation. In this paper, inositol permeability enzyme gene (iol T1), glucokinase gene (ppgk) and EIIGlc gene (pts G), were overexpressed in ZL-8 of L- lysine producing bacteria (Corynebacterium glutamicum (C.glutamicum), where glucose metabolism is slow. To study the effects of glucose transporter related genes on glucose metabolism rate, L-lysine and by-product synthesis. Compared with the original strain C.glutamicum ZL-8, Glucose consumption of recombinant bacteria C.glutamicum ZL-8/p ECXK-99E-ppgkiol T1 (increased by 76.8%) and C.glutamicum ZL-8/p DXW-8-pts G (PTS~ (Glc) system increased by 62.7%) The rate increased by 16.7% and 27.3%, respectively. The yield of L-lysine increased by 24. 7% and 16. 4%, respectively. In addition, the analysis of organic acids and amino acids showed that overexpression of different glucose transporter genes had different effects on organic acids (pyruvate, lactic acid and acetic acid) and by-product amino acids (alanine, valine, aspartic acid).
【作者單位】: 江南大學(xué)生物工程學(xué)院工業(yè)生物技術(shù)教育部重點(diǎn)實(shí)驗(yàn)室;
【基金】:江蘇省自然科學(xué)基金青年基金(BK20150149)
【分類號(hào)】:TQ922.3


本文編號(hào):2411638

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