Kinetic studies on the metabolism of short-chain fatty acids and glucose by isolated rat colonocytes.
OBJECTIVE
Although the interest in colonic mucosal metabolism of short-chain fatty acids is steadily increasing, the kinetic parameters Vmax (maximum velocity) and Km (Michaelis constant) of the complete oxidation of these acids into CO2 by colonic epithelial cells have not previously been determined.


METHODS
Isolated rat colonocytes were incubated in the presence of a concentration range of 14C-labeled acetate, propionate, butyrate, and glucose. Oxidation rates were obtained by quantifying the production of 14CO2. Vmax and Km were calculated by computer-fitting of the data to a Michaelis-Menten plot.


RESULTS
The apparent Vmax values were similar comparing acetate, propionate, and butyrate (1.114 +/- 0.061, 0.991 +/- 0.072, and 1.007 +/- 0.070 mumol/min.g, respectively), but significantly lower for glucose (0.339 +/- 0.022 mumol/min.g). The corresponding Km values were all different and in the order of butyrate (0.184 +/- 0.017 mmol/L) < propionate (0.339 +/- 0.025 mmol/L) < acetate (0.487 +/- 0.019 mmol/L) < glucose (0.777 +/- 0.051 mmol/L). In substrate competition experiments, butyrate caused a strong noncompetitive inhibition of acetate oxidation and a mixed type of inhibition of propionate oxidation. Propionate inhibited the oxidation of acetate noncompetitively and that of butyrate competitively. Acetate only slightly inhibited the oxidation of propionate and butyrate.


CONCLUSIONS
Colonic epithelial cells seem to utilize short-chain fatty acids in a preferential order of butyrate > propionate > acetate. Oxidation of propionate or acetate, however, may provide the energy needed for cellular functions if the metabolism of butyrate is impaired or the luminal supply is limited.
Clausen
M R
MR
Department of Medicine A, Rigshospitalet, University of Copenhagen, Denmark.
Mortensen
P B
PB
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism; D003065:Coenzyme A / Q000494:pharmacology; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D005232:Fatty Acids, Volatile / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D066298:In Vitro Techniques; D007657:Ketone Bodies / Q000096:biosynthesis; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D008297:Male; D010368:Pectins / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014280:Triglycerides / Q000494:pharmacology
D005232:Fatty Acids, Volatile; D007657:Ketone Bodies; D007773:Lactates; D014280:Triglycerides; D002245:Carbon Dioxide; D019344:Lactic Acid; D010368:Pectins; D000255:Adenosine Triphosphate; D005947:Glucose; C005830:tributyrin; D003065:Coenzyme A