[Effect of overexpression of nicotinic acid phosphoribosyl transferase on succinic acid production in Escherichia coli NZN111]

Sheng Wu Gong Cheng Xue Bao. 2011 Oct;27(10):1438-47.
[Article in Chinese]

Abstract

Escherichia coli strain NZN111 is a promising candidate for the fermentative production of succinate. However, because lactate dehydrogenase and pyruvate formate lyase were inactivated in NZN111, this strain had an unbalanced NADH/NAD+ ratio and could not use glucose under anaerobic conditions. In this study, a recombinant strain E. coli NZN111/pTrc99a-pncB was constructed to overexpress the nicotinic acid phosphoribosyl transferase gene (pncB). Under anaerobic conditions with the addition of 0.5 mmol/L nicotinic acid and 0.3 mmol/L isopropyl beta-D-thiogalactopyranoside (IPTG), the specific nicotinic acid phosphoribosyl transferase (NAPRTase, EC 2.4.2.11) activity in the recombinant strain was 11-fold higher than that in E. coli NZN111, the concentration of NAD(H) was increased by 3.85-fold, especially the concentration of NAD+ was increased by 5.17-fold and NADH/NAD+ was decreased from 0.640 to 0.125. The recombinant strain regained the capability of growth and glucose utilization under anaerobic conditions.

MeSH terms

  • Acetyltransferases / genetics
  • Acetyltransferases / metabolism
  • Anaerobiosis
  • Escherichia coli / classification
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Fermentation*
  • Genetic Enhancement / methods*
  • Glucose / metabolism
  • L-Lactate Dehydrogenase / genetics
  • L-Lactate Dehydrogenase / metabolism
  • NAD / metabolism
  • Nicotinamide Phosphoribosyltransferase / biosynthesis*
  • Nicotinamide Phosphoribosyltransferase / genetics
  • Succinic Acid / metabolism*

Substances

  • NAD
  • Succinic Acid
  • L-Lactate Dehydrogenase
  • Acetyltransferases
  • formate C-acetyltransferase
  • Nicotinamide Phosphoribosyltransferase
  • Glucose