Metabolic engineering of Escherichia coli to enhance acetol production from glycerol

Ruilian Yao, Qing Liu, Hongbo Hu, Thomas Keith Wood, Xuehong Zhang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Acetol, a C3 keto alcohol, is an important intermediate used to produce polyols and acrolein. To enhance acetol production from glycerol by Escherichia coli, a mutant (HJ02) was constructed by replacing the native glpK gene with the allele from E. coli Lin 43 and overexpression of yqhD, which encodes aldehyde oxidoreductase YqhD that converts methylglyoxal to acetol. Compared to the control strain without the glpK replacement, HJ02 had 5.5 times greater acetol production and a 53.4 % higher glycerol consumption rate. Then, glucose was added as a co-substrate to enhance NADPH availability and the ptsG gene was deleted in HJ02 (HJ04) to alleviate carbon catabolite repression, which led to a 30 % increase in the NADPH level and NADPH/NADP+. Consequently, HJ04 accumulated up to 1.20 g/L of acetol, which is 69.0 % higher than that of HJ02. Furthermore, the gapA gene in HJ04 was silenced by antisense RNA (HJ05) to further enhance acetol production. The acetol concentration produced by HJ05 reached 1.82 g/L, which was 2.1 and 1.5 times higher than that of HJ02 and HJ04. Real-time PCR analysis indicates that glucose catabolism was rerouted from glycolysis to the oxidative pentose phosphate pathway in HJ05.

Original languageEnglish (US)
Pages (from-to)7945-7952
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume99
Issue number19
DOIs
StatePublished - Oct 10 2015

Fingerprint

Metabolic Engineering
Glycerol
Escherichia coli
NADP
Aldehyde Oxidoreductases
Genes
Catabolite Repression
Pyruvaldehyde
Acrolein
Glucose
Antisense RNA
Pentose Phosphate Pathway
Glycolysis
acetol
Real-Time Polymerase Chain Reaction
Alleles
Alcohols

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Yao, Ruilian ; Liu, Qing ; Hu, Hongbo ; Wood, Thomas Keith ; Zhang, Xuehong. / Metabolic engineering of Escherichia coli to enhance acetol production from glycerol. In: Applied Microbiology and Biotechnology. 2015 ; Vol. 99, No. 19. pp. 7945-7952.
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abstract = "Acetol, a C3 keto alcohol, is an important intermediate used to produce polyols and acrolein. To enhance acetol production from glycerol by Escherichia coli, a mutant (HJ02) was constructed by replacing the native glpK gene with the allele from E. coli Lin 43 and overexpression of yqhD, which encodes aldehyde oxidoreductase YqhD that converts methylglyoxal to acetol. Compared to the control strain without the glpK replacement, HJ02 had 5.5 times greater acetol production and a 53.4 {\%} higher glycerol consumption rate. Then, glucose was added as a co-substrate to enhance NADPH availability and the ptsG gene was deleted in HJ02 (HJ04) to alleviate carbon catabolite repression, which led to a 30 {\%} increase in the NADPH level and NADPH/NADP+. Consequently, HJ04 accumulated up to 1.20 g/L of acetol, which is 69.0 {\%} higher than that of HJ02. Furthermore, the gapA gene in HJ04 was silenced by antisense RNA (HJ05) to further enhance acetol production. The acetol concentration produced by HJ05 reached 1.82 g/L, which was 2.1 and 1.5 times higher than that of HJ02 and HJ04. Real-time PCR analysis indicates that glucose catabolism was rerouted from glycolysis to the oxidative pentose phosphate pathway in HJ05.",
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Metabolic engineering of Escherichia coli to enhance acetol production from glycerol. / Yao, Ruilian; Liu, Qing; Hu, Hongbo; Wood, Thomas Keith; Zhang, Xuehong.

In: Applied Microbiology and Biotechnology, Vol. 99, No. 19, 10.10.2015, p. 7945-7952.

Research output: Contribution to journalArticle

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AB - Acetol, a C3 keto alcohol, is an important intermediate used to produce polyols and acrolein. To enhance acetol production from glycerol by Escherichia coli, a mutant (HJ02) was constructed by replacing the native glpK gene with the allele from E. coli Lin 43 and overexpression of yqhD, which encodes aldehyde oxidoreductase YqhD that converts methylglyoxal to acetol. Compared to the control strain without the glpK replacement, HJ02 had 5.5 times greater acetol production and a 53.4 % higher glycerol consumption rate. Then, glucose was added as a co-substrate to enhance NADPH availability and the ptsG gene was deleted in HJ02 (HJ04) to alleviate carbon catabolite repression, which led to a 30 % increase in the NADPH level and NADPH/NADP+. Consequently, HJ04 accumulated up to 1.20 g/L of acetol, which is 69.0 % higher than that of HJ02. Furthermore, the gapA gene in HJ04 was silenced by antisense RNA (HJ05) to further enhance acetol production. The acetol concentration produced by HJ05 reached 1.82 g/L, which was 2.1 and 1.5 times higher than that of HJ02 and HJ04. Real-time PCR analysis indicates that glucose catabolism was rerouted from glycolysis to the oxidative pentose phosphate pathway in HJ05.

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