Jacobson B S, Smith B N, Epstein S, Laties G G
J Gen Physiol. 1970 Jan;55(1):1-17. doi: 10.1085/jgp.55.1.1.
Isotope discrimination is a common feature of biosynthesis in nature, with the result that different classes of carbon compounds frequently display different (13)C/(12)C ratios. The (13)C/(12)C ratio of lipid in potato tuber tissue is considerably lower than that for starch or protein. We have collected respiratory CO(2) 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 (13)C/(12)C ratio of the evolved CO(2) was determined for each period, and compared with the (13)C/(12)C ratios of the major tissue metabolites. In the first hours the carbon isotope ratio of the CO(2) 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 alpha-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 CO(2) may be obscured by fractionation attending the refixation of CO(2) during respiration, and by CO(2) arising from dissolved CO(2) and bicarbonate preexisting in the tuber. Means are described for coping with both pitfalls.
同位素分馏是自然界生物合成的一个常见特征,结果是不同种类的碳化合物常常呈现出不同的碳-13/碳-12比率。马铃薯块茎组织中脂质的碳-13/碳-12比率明显低于淀粉或蛋白质的该比率。我们从切割之时起连续24小时收集马铃薯圆片呼出的二氧化碳,在这段时间内呼吸速率提高了3至5倍。测定了每个时间段呼出二氧化碳的碳-13/碳-12比率,并与主要组织代谢物的碳-13/碳-12比率进行比较。在最初几个小时,二氧化碳的碳同位素比率与脂质的比率相符。随着时间推移,该比率接近淀粉或蛋白质的典型比率。已经估算出每个时间点脂质和碳水化合物对总呼吸作用的贡献。结合其他观察结果推断,基础呼吸或初始呼吸代表脂质代谢,可能是长链脂肪酸的α-氧化,而增强的呼吸代表碳水化合物糖酵解产物的传统三羧酸循环氧化。呼吸过程中二氧化碳重新固定引起的分馏以及块茎中预先存在的溶解二氧化碳和碳酸氢盐产生的二氧化碳可能会掩盖呼出二氧化碳的真实同位素组成。文中描述了应对这两个问题的方法。