Small F J, Ensign S A
Department of Chemistry and Biochemistry, Utah State University, Logan 84322-0300, USA.
J Bacteriol. 1995 Nov;177(21):6170-5. doi: 10.1128/jb.177.21.6170-6175.1995.
Evidence for a requirement for CO2 in the productive metabolism of aliphatic alkenes and epoxides by the propylene-oxidizing bacterium Xanthobacter strain Py2 is presented. In the absence of CO2, whole-cell suspensions of propylene-grown cells catalyzed the isomerization of propylene oxide (epoxypropane) to acetone. In the presence of CO2, no acetone was produced. Acetone was not metabolized by suspensions of propylene-grown cells, in either the absence or presence of CO2. The degradation of propylene and propylene oxide by propylene-grown cells supported the fixation of 14CO2 into cell material, and the time course of 14C fixation correlated with the time course of propylene and propylene oxide degradation. The degradation of glucose and propionaldehyde by propylene-grown or glucose-grown cells did not support significant 14CO2 fixation. With propylene oxide as the substrate, the concentration dependence of 14CO2 fixation exhibited saturation kinetics, and at saturation, 0.9 mol of CO2 was fixed per mol of propylene oxide consumed. Cultures grown with propylene in a nitrogen-deficient medium supplemented with NaH13CO3 specifically incorporated 13C label into the C-1 (major labeled position) and C-3 (minor labeled position) carbon atoms of the endogenous storage compound poly-beta-hydroxybutyrate. No specific label incorporation was observed when cells were cultured with glucose or n-propanol as a carbon source. The depletion of CO2 from cultures grown with propylene, but not glucose or n-propanol, inhibited bacterial growth. We propose that propylene oxide metabolism in Xanthobacter strain Py2 proceeds by terminal carboxylation of an isomerization intermediate, which, in the absence of CO2, is released as acetone.
本文提供了关于氧化丙烯细菌黄杆菌属菌株Py2在脂肪族烯烃和环氧化物的生产性代谢中对二氧化碳需求的证据。在没有二氧化碳的情况下,以丙烯培养的细胞的全细胞悬浮液催化环氧丙烷(氧化丙烯)异构化为丙酮。在有二氧化碳的情况下,不产生丙酮。无论有无二氧化碳,以丙烯培养的细胞悬浮液都不能代谢丙酮。以丙烯培养的细胞对丙烯和环氧丙烷的降解支持将(^{14}CO_2)固定到细胞物质中,并且(^{14}C)固定的时间进程与丙烯和环氧丙烷降解的时间进程相关。以丙烯培养或葡萄糖培养的细胞对葡萄糖和丙醛的降解不支持显著的(^{14}CO_2)固定。以环氧丙烷为底物时,(^{14}CO_2)固定的浓度依赖性表现出饱和动力学,在饱和时,每消耗1摩尔环氧丙烷固定0.9摩尔二氧化碳。在补充有(NaH^{13}CO_3)的缺氮培养基中用丙烯培养的培养物将(^{13}C)标记特异性地掺入内源性储存化合物聚-β-羟基丁酸酯的C-1(主要标记位置)和C-3(次要标记位置)碳原子中。当用葡萄糖或正丙醇作为碳源培养细胞时,未观察到特异性标记掺入。用丙烯培养但不是用葡萄糖或正丙醇培养的培养物中二氧化碳的消耗会抑制细菌生长。我们提出,黄杆菌属菌株Py2中的环氧丙烷代谢是通过异构化中间体的末端羧化进行的,在没有二氧化碳的情况下,该中间体以丙酮的形式释放。