Transcription levels of key metabolic genes are the cause for different glucose utilization pathways in E. coli B (BL21) and E. coli K (JM109).
Acetate accumulation is a common problem observed in aerobic high cell density cultures of Escherichia coli. It has been hypothesized in previous reports that the glyoxylate shunt is active in E. coli BL21, the low acetate producer, and inactive in E. coli JM109, the high acetate producer. This hypothesis was further strengthened by incorporating 13C from uniformly labeled glucose into TCA cycle intermediates. Using northern blot analyses, the current report demonstrates that the reason for the inactivity of the glyoxylate pathway in E. coli JM109 is the no apparent transcription of isocitrate lyase (aceA) and malate synthase (aceB), and transcription of the isocitrate lyase repressor (iclR). The reverse is seen in E. coli BL21 where the glyoxylate pathway is active due to constitutive transcription of aceA and aceB and no transcription of the iclR. In addition, there is a difference between the two strains in the transcription of the acetyl-CoA synthetase (acs), phosphotransacetylase-acetate kinase (pta-ackA) pathway, and pyruvate oxidase (poxB), pathway. The transcript of acs is higher in E. coli BL21 and lower in the E. coli JM109, while the reverse is true for poxB transcription.
Phue
Je-Nie
JN
Biotechnology Unit, NIH, NIDDK, Building 14A Rm 173, Bethesda, MD 20892, USA.
Shiloach
Joseph
J
D000084:Acetate Kinase / Q000235:genetics; D000106:Acetate-CoA Ligase / Q000235:genetics; D000085:Acetates / Q000378:metabolism; D002952:Citric Acid Cycle / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D005285:Fermentation; D005947:Glucose / Q000378:metabolism*; D006038:Glyoxylates / Q000378:metabolism; D010709:Phosphate Acetyltransferase / Q000235:genetics; D011771:Pyruvate Oxidase / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic
D000085:Acetates; D006038:Glyoxylates; D012333:RNA, Messenger; D011771:Pyruvate Oxidase; D010709:Phosphate Acetyltransferase; D000084:Acetate Kinase; D000106:Acetate-CoA Ligase; D005947:Glucose; C031150:glyoxylic acid