Hopper D J, Chapman P J, Dagley S
Department of Biochemistry, University of Minnesota, St. Paul, Minnesota 55101.
J Bacteriol. 1970 Dec;104(3):1197-202. doi: 10.1128/jb.104.3.1197-1202.1970.
Extracts of a fluorescent species of Pseudomonas grown with m-cresol, degrade gentisic acid without isomerization of the ring-fission compound, maleylpyruvate, to give eventually d-malate and pyruvate. d-Malate is also a growth substrate. l-Malate but not d-malate is oxidized by a particulate enzyme not requiring nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP). NAD- or NADP-linked malate dehydrogenases are absent but cells contain an NADP-dependent l-malic enzyme. Exposure of cells to exogenous d-malate induces an NAD-dependent d-malic enzyme, not present when d-malate is formed endogenously. Succinate- or m-cresol-grown cells, containing no d-malic enzyme, rapidly oxidize d-malate in the presence of chloramphenicol at a concentration suffient to inhibit protein synthesis. An NADP-dependent cell-free system, prepared from succinate-grown cells which oxidized d-malate, is described.
用间甲酚培养的荧光假单胞菌提取物可降解龙胆酸,且环裂解化合物马来酰丙酮酸不会发生异构化,最终生成d-苹果酸和丙酮酸。d-苹果酸也是一种生长底物。l-苹果酸可被一种不依赖烟酰胺腺嘌呤二核苷酸(NAD)或烟酰胺腺嘌呤二核苷酸磷酸(NADP)的颗粒酶氧化,但d-苹果酸不行。细胞中不存在NAD或NADP连接的苹果酸脱氢酶,但含有一种NADP依赖的l-苹果酸酶。将细胞暴露于外源性d-苹果酸会诱导产生一种NAD依赖的d-苹果酸酶,而当d-苹果酸内源性形成时该酶不存在。在含有足以抑制蛋白质合成浓度的氯霉素存在下,不含d-苹果酸酶的琥珀酸盐或间甲酚培养的细胞能迅速氧化d-苹果酸。本文描述了一种由氧化d-苹果酸的琥珀酸盐培养细胞制备的NADP依赖的无细胞体系。