Lundgren D W, Fales H M
J Biol Chem. 1980 May 25;255(10):4481-6.
Incubation of 2-[14C]pyrrolidone with sliced rat liver and analysis of the incubation medium by silica gel chromatography revealed that 2-[14C]pyrrolidone is metabolized to an unknown. It was previously shown by Lundgren and Hankins ((Lundgren, D.W, and Hankins, J. (1978) J. Biol. Chem. 253, 7130-7133) that slices of rat liver readily synthesized 2-pyrrolidone from putrescine. The unknown metabolite was partially purified by methanol/chloroform extraction, activated charcoal column chromatography, and two-dimensional thin layer chromatography on silica gel plates. The 2-pyrrolidone metabolite was derivatized with bis(trimethylsilyl)trifluroacetamide and analyzed by gas chromatography-mass spectrometry. The mass of the molecular ion (245) and fragment ions suggests that the 2-pyrrolidone metabolite is 5-hydroxy-2-pyrrolidone. The mass spectrum of synthetic 5-hydroxy-2-pyrrolidone was identical to that of the unknown metabolite. Synthetic 5-hydroxy-2-[3H]pyrrolidone co-chromatographed on silica gel sheets with the unknown 2-[14C]pyrrolidone metabolite obtained directly from incubation media. Under appropriate conditions (pH 7.5, no acid treatment of medium), putrescine is metabolized to 5-hydroxy-2-pyrrolidone via 2-pyrrolidone. Several effector compounds, but not necessarily the same ones, inhibit or enhance, or both, the conversion of putrescine to 2-pyrrolidone and of 2-pyrrolidone to 5-hydroxy-2-pyrrolidone. This is the first demonstration of the biosynthesis of 5-hydroxy-2-pyrrolidone.
将2-[¹⁴C]吡咯烷酮与大鼠肝脏切片一起孵育，并通过硅胶色谱法对孵育培养基进行分析，结果显示2-[¹⁴C]吡咯烷酮代谢生成了一种未知物。伦德格伦和汉金斯此前曾表明（伦德格伦，D.W.，和汉金斯，J.（1978年）《生物化学杂志》253卷，7130 - 7133页），大鼠肝脏切片能轻易地从腐胺合成2-吡咯烷酮。该未知代谢物通过甲醇/氯仿萃取、活性炭柱色谱法以及硅胶板上的二维薄层色谱法进行了部分纯化。2-吡咯烷酮代谢物用双（三甲基硅基）三氟乙酰胺进行衍生化，并通过气相色谱 - 质谱联用仪进行分析。分子离子（245）和碎片离子的质量表明该2-吡咯烷酮代谢物是5-羟基-2-吡咯烷酮。合成的5-羟基-2-吡咯烷酮的质谱与未知代谢物的质谱相同。合成的5-羟基-2-[³H]吡咯烷酮与直接从孵育培养基中获得的未知2-[¹⁴C]吡咯烷酮代谢物在硅胶板上共色谱。在适当条件下（pH 7.5，不对培养基进行酸处理），腐胺通过2-吡咯烷酮代谢生成5-羟基-2-吡咯烷酮。几种效应化合物，但不一定是相同的化合物，会抑制或增强，或同时抑制和增强腐胺向2-吡咯烷酮的转化以及2-吡咯烷酮向5-羟基-2-吡咯烷酮的转化。这是首次证明5-羟基-2-吡咯烷酮的生物合成。
