The fed-batch production of a thermophilic 2-deoxyribose-5-phosphate aldolase (DERA) in Escherichia coli by exponential feeding strategy control

Appl Biochem Biotechnol. 2010 Nov;162(5):1423-34. doi: 10.1007/s12010-010-8924-1. Epub 2010 Mar 15.

Abstract

2-Deoxyribose-5-phosphate aldolase (DERA) catalyzes a sequential aldol reaction useful in synthetic chemistry. In this work, the effect of a feeding strategy on the production of a thermophilic DERA was investigated in fed-batch cultures of recombinant Escherichia coli BL21 (pET303-DERA008). The predetermined specific growth rate (micro(set)) was evaluated at 0.20, 0.15, and 0.10 h(-1), respectively. The DERA concentration and volumetric productivity were associated with micro(set). The cells synthesized the enzyme most efficiently at micro(set) = 0.15 h(-1). The maximum enzyme concentration (5.12 g/L) and total volumetric productivity (0.256 g L(-1) h(-1)) obtained were over 10 and five times higher than that from traditional batch cultures. Furthermore, the acetate concentration remained at a relatively low level, less than 0.4 g/L, under this condition which would not inhibit cell growth and target protein expression. Thus, a specific growth rate control strategy has been successfully applied to induce fed-batch cultures for the maximal production of the thermophilic 2-deoxyribose-5-phosphate aldolase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde-Lyases / biosynthesis*
  • Bioreactors / microbiology
  • Biotechnology / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / enzymology*
  • Fermentation / physiology*
  • Temperature*

Substances

  • Aldehyde-Lyases
  • deoxyribose-phosphate aldolase