The Prevalence of Carbon-13 in Respiratory Carbon Dioxide As an Indicator of the Type of Endogenous Substrate

Abstract

Isotope discrimination is a common feature of biosynthesis in nature, with the result that different classes of carbon compounds frequently display different 13C/12C ratios. The 13C/12C ratio of lipid in potato tuber tissue is considerably lower than that for starch or protein. We have collected respiratory CO2 from potato discs in successive periods through 24 hr from the time of cutting—an interval in which the respiration rate rises 3–5-fold. The 13C/12C ratio of the evolved CO2 was determined for each period, and compared with the 13C/12C ratios of the major tissue metabolites. In the first hours the carbon isotope ratio of the CO2 matches that of lipid. With time, the ratio approaches that typical of starch or protein. An estimation has been made of the contribution of lipid and carbohydrate to the total respiration at each juncture. In connection with additional observations, it was deduced that the basal, or initial, respiration represents lipid metabolism—possibly the α-oxidation of long chain fatty acids—while the developed repiration represents conventional tricarboxylic acid cycle oxidation of the products of carbohydrate glycolysis. The true isotopic composition of the respiratory CO2 may be obscured by fractionation attending the refixation of CO2 during respiration, and by CO2 arising from dissolved CO2 and bicarbonate preexisting in the tuber. Means are described for coping with both pitfalls.