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Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP)

หน่วยงาน Nanyang Technological University, Singapore

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ชื่อเรื่อง : Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP)
นักวิจัย : Chong, Huiqing , Yeow, Jianwei , Wang, Ivy , Song, Hao , Jiang, Rongrong
คำค้น : DRNTU::Engineering::Chemical engineering::Biochemical engineering.
หน่วยงาน : Nanyang Technological University, Singapore
ผู้ร่วมงาน : Isalan, Mark
ปีพิมพ์ : 2556
อ้างอิง : Chong, H., Yeow, J., Wang, I., Song, H., & Jiang, R. (2013). Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP). PLoS ONE, 8(10), e77422-. , 1932-6203 , http://hdl.handle.net/10220/18294 , http://dx.doi.org/10.1371/journal.pone.0077422
ที่มา : -
ความเชี่ยวชาญ : -
ความสัมพันธ์ : PLoS ONE
ขอบเขตของเนื้อหา : -
บทคัดย่อ/คำอธิบาย :

The presence of acetate exceeding 5 g/L is a major concern during E. coli fermentation due to its inhibitory effect on cell growth, thereby limiting high-density cell culture and recombinant protein production. Hence, engineered E. coli strains with enhanced acetate tolerance would be valuable for these bioprocesses. In this work, the acetate tolerance of E. coli was much improved by rewiring its global regulator cAMP receptor protein (CRP), which is reported to regulate 444 genes. Error-prone PCR method was employed to modify crp and the mutagenesis libraries (~3×106) were subjected to M9 minimal medium supplemented with 5–10 g/L sodium acetate for selection. Mutant A2 (D138Y) was isolated and its growth rate in 15 g/L sodium acetate was found to be 0.083 h-1, much higher than that of the control (0.016 h-1). Real-time PCR analysis via OpenArray® system revealed that over 400 CRP-regulated genes were differentially expressed in A2 with or without acetate stress, including those involved in the TCA cycle, phosphotransferase system, etc. Eight genes were chosen for overexpression and the overexpression of uxaB was found to lead to E. coli acetate sensitivity.

บรรณานุกรม :
Chong, Huiqing , Yeow, Jianwei , Wang, Ivy , Song, Hao , Jiang, Rongrong . (2556). Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP).
    กรุงเทพมหานคร : Nanyang Technological University, Singapore.
Chong, Huiqing , Yeow, Jianwei , Wang, Ivy , Song, Hao , Jiang, Rongrong . 2556. "Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP)".
    กรุงเทพมหานคร : Nanyang Technological University, Singapore.
Chong, Huiqing , Yeow, Jianwei , Wang, Ivy , Song, Hao , Jiang, Rongrong . "Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP)."
    กรุงเทพมหานคร : Nanyang Technological University, Singapore, 2556. Print.
Chong, Huiqing , Yeow, Jianwei , Wang, Ivy , Song, Hao , Jiang, Rongrong . Improving acetate tolerance of Escherichia coli by rewiring Its global regulator cAMP receptor protein (CRP). กรุงเทพมหานคร : Nanyang Technological University, Singapore; 2556.