Smith R G, Gauthier D A, Dennis D T, Turpin D H
Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4.
Plant Physiol. 1992 Apr;98(4):1233-8. doi: 10.1104/pp.98.4.1233.
Leucoplasts were isolated from the endosperm of developing castor (Ricinis communis) endosperm using a discontinuous Percoll gradient. The rate of fatty acid synthesis was highest when malate was the precursor, at 155 nanomoles acetyl-CoA equivalents per milligram protein per hour. Pyruvate and acetate also were precursors of fatty acid synthesis, but the rates were approximately 4.5 and 120 times less, respectively, than when malate was the precursor. When acetate was supplied to leucoplasts, exogenous ATP, NADH, and NADPH were required to obtain maximal rates of fatty acid synthesis. In contrast, the incorporation of malate and pyruvate into fatty acids did not require a supply of exogenous reductant. Further, the incorporation of radiolabel into fatty acids by leucoplasts supplied with radiolabeled malate, pyruvate, or acetate was reduced upon coincubation with cold pyruvate or malate. The data suggest that malate and pyruvate may be good in vivo sources of carbon for fatty acid synthesis and that, in these preparations, leucoplast fatty acid synthesis may be limited by activity at or downstream of the acetyl-CoA carboxylase reaction.
利用不连续的Percoll梯度从发育中的蓖麻(Ricinis communis)胚乳中分离出白色体。当以苹果酸为前体时,脂肪酸合成速率最高,为每毫克蛋白质每小时155纳摩尔乙酰辅酶A当量。丙酮酸和乙酸盐也是脂肪酸合成的前体,但速率分别比以苹果酸为前体时低约4.5倍和120倍。当向白色体供应乙酸盐时,需要外源ATP、NADH和NADPH才能获得脂肪酸合成的最大速率。相比之下,苹果酸和丙酮酸掺入脂肪酸不需要供应外源还原剂。此外,与冷丙酮酸或苹果酸共同孵育后,用放射性标记的苹果酸、丙酮酸或乙酸盐供应的白色体将放射性标记掺入脂肪酸的量减少。数据表明,苹果酸和丙酮酸可能是体内脂肪酸合成良好的碳源,并且在这些制剂中,白色体脂肪酸合成可能受到乙酰辅酶A羧化酶反应或其下游活性的限制。