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生物化学 1937

柠檬酸在动物组织中间代谢中的作用

汉斯·克雷布斯与威廉·约翰逊

细胞如何燃烧燃料:一圈自我再生的反应,把食物变成 CO₂ 与可用的能量。

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In depth · the introduction

你每一次呼吸,都在喂养细胞里一座小小的化学旋转木马——一圈反应,燃烧你的食物,却从不停转。

核心想法

细胞靠用你吸入的氧来「燃烧」食物获得能量。汉斯·克雷布斯发现,这场燃烧不是一条笔直的流水线,而是一个环。一小块食物的片段,搭上一个载体,绕着一圈八步的环走一遭,把碳以你呼出的 CO₂ 的形式甩掉,沿途交出能量——然后载体被重建,准备好抓住下一个片段。

这环一圈又一圈地转。因为每一圈都重建出自己的起始物,所以环里的化学物质,任何时候都只需极少量——它们是机器,不是燃料。这个自我更新的环,就是柠檬酸循环,也叫克雷布斯循环。

它是如何诞生的

克雷布斯是位德国出生的生物化学家,1933 年被逐出德国,在英格兰重建了自己的事业。他用绞碎的鸽胸肌——一种呼吸很猛的组织——做实验,注意到一件怪事:加入一小撮柠檬酸,竟让肌肉耗掉远超那一撮所能解释的氧。柠檬酸是被用过又重造,而非被烧尽。

把别的化学家留下的线索拼到一起,克雷布斯意识到,这些酸构成了一个自我更新的圆环。他把结果投给顶尖的《自然》杂志——却被退稿。它最终发表在一份较小的期刊《酶学》上。十六年后,1953 年,正是这项工作为他赢得了诺贝尔奖。

它为何重要

这循环,是几乎每一种用氧的生命——一个细菌、一棵树、还有你——从食物里取出能量的方式。它是代谢的大配电盘:糖、脂肪、蛋白质,都汇进同一个环里被燃烧。把它画清楚,就把生命的化学从一团各自为政的反应,变成了一个单一、可理解的系统——一个医学与生物学至今每天都在依赖的系统。

一个可以想象的画面

想象一座水车。溪流——你的食物——泼到水车上;水车转动、做功,把能量收下;用过的水在离开时泼洒而出,就像你呼出的 CO₂。最关键的是,水车本身并不被消耗——它转一圈又回到顶上,等着下一波水花。循环里的那些酸就是水车;只需一点点,因为它会不停地转回来。

可交互的柠檬酸循环:八种中间物排成一圈(草酰乙酸、柠檬酸、异柠檬酸、α-酮戊二酸、琥珀酰辅酶A、琥珀酸、延胡索酸、苹果酸);滑块沿八步酶促反应推进,圆心显示当前分子的碳原子数,旁边实时累计本圈已产出的 CO₂、NADH、FADH₂ 与 GTP,专家面板给出每圈的固定总数与约 10 个 ATP 的产率。

它的位置

这循环是细胞产能的中间一幕:在它之前,食物被拆成小片段;在它之后,被它充满的能量载体(NADH 及其同类)驱动着细胞的主发电厂。它与本馆中其他关于生命分子的发现并肩而立——从霍奇金与赫胥黎的带电神经,到沃森与克里克的 DNA——同属二十世纪「把生命读成化学」的那项事业。

The original document
Original source text
H. A. Krebs & W. A. Johnson · Enzymologia 4 (1937): 148–156 · University of Sheffield
The paper sets out to explain how animal tissues oxidize carbohydrate all the way to carbon dioxide and water. The experimental material is minced pigeon breast muscle — a tissue chosen for its exceptionally vigorous respiration — suspended in a manometer that reads its uptake of oxygen as small amounts of various organic acids are added.
The decisive observation is that the effect of citrate is catalytic. Adding a small, sub-stoichiometric amount of citric acid raises the muscle's total oxygen consumption by far more than the oxidation of that citrate alone could account for. The citrate is therefore not consumed but regenerated — it cycles. The same catalytic potency had been seen for a family of di- and tricarboxylic acids (succinate, fumarate, malate, oxaloacetate, α-ketoglutarate, citrate) that earlier workers had each studied separately.
Krebs's synthesis is to close these fragments into a single ring. A substance derived from carbohydrate condenses with the four-carbon oxaloacetate to form six-carbon citric acid; through a sequence of oxidations and two losses of CO₂ the molecule is whittled back down to oxaloacetate, which is then free to condense again. Because oxaloacetate is continuously regenerated, only a catalytic trace is needed — exactly matching the experiment.
[ … ]
The note is the cycle's first full statement, not its finished biochemistry: the identity of the two-carbon unit that actually enters the ring — the acetyl group carried by coenzyme A — was unknown in 1937 and was established only after Lipmann's later discovery of coenzyme A. Submitted first to the journal Nature, the manuscript was declined, and appeared instead in Enzymologia.
University of Sheffield · 1937