Accumulation of methylglyoxal in anaerobically grown Escherichia coli and its detoxification by expression of the Pseudomonas putida glyoxalase I gene.
Anaerobic glycerol fermentation by Escherichia coli strains expressing genes from the Klebsiella pneumoniae dha regulon showed that cell growth and 1,3-propanediol (1,3-PD) production are significantly inhibited when 5 g/L or higher of glycerol is initially present. One reason for this inhibition may be methylglyoxal (MG) accumulation. Assays of both intracellular and extracellular MG levels indicated an accumulation of MG in anaerobic glycerol fermentation of transgenic E. coli. Pseudomonas putida glyoxalase I was expressed in the transgenic E. coli to enhance MG detoxification. The activity of glyoxalase I in the transgenic E. coli with the P. putida glyoxalase I under anaerobic conditions was 12-fold higher than that in the control cells. Compared to the control cells, the transgenic cells with the P. putida glyoxalase I displayed a reduction of 35-43% in intracellular MG and a decrease of 30% in extracellular MG. These decreases were statistically significant (P>94). Furthermore, the expression of the P. putida glyoxalase I in the transgenic E. coli markedly improved cell growth and resulted in a 50% increase in 1,3-PD production.
Zhu
M M
MM
Department of Chemical Engineering, University of Wisconsin-Madison, Madison, WI 53706-1691, USA. marie_zhu@groton.pfizer.com
Skraly
F A
FA
Cameron
D C
DC
D000693:Anaerobiosis; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D015870:Gene Expression; D005798:Genes, Bacterial; D008658:Inactivation, Metabolic; D007791:Lactoylglutathione Lyase / Q000235:genetics; D011409:Propylene Glycols / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics; D011765:Pyruvaldehyde / Q000378:metabolism*
D017931:DNA Primers; D011409:Propylene Glycols; C041787:1,3-propanediol; D011765:Pyruvaldehyde; D007791:Lactoylglutathione Lyase