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一种节杆菌对L-苏氨酸的利用。“氨基丙酮合酶”的一种新的分解代谢作用。

Utilization of L-threonine by a species of Arthrobacter. A novel catabolic role for "aminoacetone synthase".

作者信息

McGilvray D, Morris J G

出版信息

Biochem J. 1969 May;112(5):657-71. doi: 10.1042/bj1120657.

DOI:10.1042/bj1120657
PMID:5821726
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1187769/
Abstract
  1. A species of Arthrobacter (designated Arthrobacter 9759) was isolated from soil by its ability to grow aerobically on l-threonine as sole source of carbon atoms, nitrogen atoms and energy; the organism also grew well on other sources of carbon atoms including glycine, but no growth was obtainable on aminoacetone or dl-1-aminopropan-2-ol. 2. During growth on threonine, (14)C from l-[U-(14)C]threonine was rapidly incorporated into glycine and citrate, and thereafter into serine, alanine, aspartate and glutamate. 3. With extracts of threonine-grown cells supplied with l-[U-(14)C]threonine, evidence was obtained of the NAD and CoA-dependent catabolism of l-threonine to produce acetyl-CoA plus glycine. Short-term incorporation studies in which [2-(14)C]acetate and [2-(14)C]glycine were supplied (a) to cultures growing on threonine, and (b) to extracts of threonine-grown cells, showed that the acetyl-CoA was metabolized via the tricarboxylic acid cycle and glyoxylate cycle whereas the glycine was converted into pyruvate via the folate-dependent ;serine pathway'. 4. The threonine-grown organism contained ;biosynthetic' threonine dehydratase and a potent NAD-linked l-threonine dehydrogenase but possessed no l-threonine aldolase activity. 5. Evidence was obtained that the acetyl-CoA and glycine produced from l-threonine had their immediate origin in the alpha-amino-beta-oxobutyrate formed by the threonine dehydrogenase; the CoA-dependent cleavage of this compound was catalysed by an alpha-amino-beta-oxobutyrate CoA-ligase, which was identified with ;aminoacetone synthase'. A continuous spectrophotometric assay of this enzyme was developed, and it was found to be inducibly synthesized only during growth on threonine and not during growth on acetate plus glycine. 6. By using a reconstituted mixture of separately purified l-threonine dehydrogenase and alpha-amino-beta-oxobutyrate CoA-ligase (i.e. ;aminoacetone synthase'), l-[U-(14)C]threonine was broken down to [(14)C]glycine plus [(14)C]acetyl-CoA (trapped as [(14)C]citrate). 7. There was no evidence of aminoacetone metabolism by Arthrobacter 9759 even though a small amount of this amino ketone appeared in the culture medium during growth on threonine.
摘要
  1. 从土壤中分离出一种节杆菌(命名为节杆菌9759),它能够以L-苏氨酸作为唯一的碳原子、氮原子和能量来源进行有氧生长;该生物体在包括甘氨酸在内的其他碳原子来源上也生长良好,但在氨基丙酮或dl-1-氨基丙醇上无法生长。2. 在以苏氨酸为碳源生长期间,L-[U-(14)C]苏氨酸中的(14)C迅速掺入甘氨酸和柠檬酸中,随后掺入丝氨酸、丙氨酸、天冬氨酸和谷氨酸中。3. 用L-[U-(14)C]苏氨酸供给以苏氨酸生长的细胞提取物,获得了L-苏氨酸经NAD和辅酶A依赖的分解代谢产生乙酰辅酶A和甘氨酸的证据。短期掺入研究中,分别向以苏氨酸生长的培养物和以苏氨酸生长的细胞提取物中供给[2-(14)C]乙酸盐和[2-(14)C]甘氨酸,结果表明乙酰辅酶A通过三羧酸循环和乙醛酸循环进行代谢,而甘氨酸通过依赖叶酸的“丝氨酸途径”转化为丙酮酸。4. 以苏氨酸生长的生物体含有“生物合成型”苏氨酸脱水酶和一种高效的NAD连接的L-苏氨酸脱氢酶,但不具有L-苏氨酸醛缩酶活性。5. 有证据表明,L-苏氨酸产生的乙酰辅酶A和甘氨酸直接来源于苏氨酸脱氢酶形成的α-氨基-β-氧代丁酸;该化合物的辅酶A依赖型裂解由α-氨基-β-氧代丁酸辅酶A连接酶催化,该酶与“氨基丙酮合酶”一致。开发了一种该酶的连续分光光度测定法,发现它仅在以苏氨酸生长期间可诱导合成,而在以乙酸盐加甘氨酸生长期间不合成。6. 通过使用分别纯化的L-苏氨酸脱氢酶和α-氨基-β-氧代丁酸辅酶A连接酶(即“氨基丙酮合酶”)的重组混合物,L-[U-(14)C]苏氨酸被分解为[(14)C]甘氨酸加[(14)C]乙酰辅酶A(捕获为[(14)C]柠檬酸)。7. 即使在以苏氨酸生长期间培养基中出现少量这种氨基酮,也没有证据表明节杆菌9759对氨基丙酮进行代谢。

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本文引用的文献

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