Department of Civil Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802.
Appl Environ Microbiol. 1985 Sep;50(3):573-9. doi: 10.1128/aem.50.3.573-579.1985.
Pathways of glucose catabolism, potentially operational in six strains of obligately aerobic, acidophilic bacteria, including Acidiphilium cryptum strain Lhet2, were investigated by short-term radiorespirometry and enzyme assays. Short-term radiorespirometry was conducted at pH 3.0 with specifically labeled [C]glucose. The high rate and yield of C-1 oxidized to CO(2) indicated that the Entner-Doudoroff, pentose phosphate, or both pathways were operational in all strains. Apparent nonequivalent yields of CO(2) from C-1 and estimated CO(2) from C-4 (C-1 > C-4) were suggestive of simultaneous glucose catabolism by both pathways in all strains tested. Variation in the relative contribution of the two pathways of glucose catabolism appears to account for observed strain differences. Calculation of the actual percent pathway participation was not feasible. Enzyme assays were completed with crude extracts of glucose-grown cells to substantiate the results obtained by radiorespirometry. The key enzymes of the pentose phosphate pathway (6-phosphogluconate dehydrogenase) and the Entner-Doudoroff pathway (2-keto-3-deoxy-6-phosphogluconate aldolase and 6-phosphogluconate dehydrase) were present in all strains examined (PW2, Lhet2, KLB, OP, and QBP). However, none of the strains exhibited detectable levels of the key enzyme of the Embden-Meyerhof-Parnas pathway, 6-phosphofructokinase. All strains contained glucose-6-phosphate dehydrogenase and fructose bisphosphate aldolase. The results of the enzyme study supported the contention that the pentose phosphate and Entner-Doudoroff pathways are operational for glucose catabolism in the acidophilic heterotrophs, and that the Embden-Meyerhof-Parnas pathway is apparently absent.
通过短期放射性呼吸测量法和酶分析,研究了 6 株严格需氧嗜酸细菌(包括嗜酸嗜酸菌 Lhet2 株)中葡萄糖分解代谢的途径,这些细菌潜在地起作用。在 pH 值为 3.0 时,用特定标记的 [C]葡萄糖进行短期放射性呼吸测量。C-1 氧化为 CO(2)的高速度和高产量表明,所有菌株都可以利用 Entner-Doudoroff、磷酸戊糖或两者途径进行葡萄糖分解代谢。从 C-1 产生的 CO(2)的明显不等效产量和估计的 C-4 产生的 CO(2)(C-1>C-4)表明,所有测试菌株都同时通过两种途径进行葡萄糖分解代谢。葡萄糖分解代谢的两种途径的相对贡献的变化似乎解释了观察到的菌株差异。计算实际的途径参与百分比是不可行的。用葡萄糖生长细胞的粗提物完成酶分析,以证实放射性呼吸测量法获得的结果。磷酸戊糖途径的关键酶(6-磷酸葡萄糖酸脱氢酶)和 Entner-Doudoroff 途径(2-酮-3-脱氧-6-磷酸葡萄糖醛酸醛缩酶和 6-磷酸葡萄糖脱水酶)存在于所有被检查的菌株(PW2、Lhet2、KLB、OP 和 QBP)中。然而,没有一种菌株表现出 Embden-Meyerhof-Parnas 途径的关键酶 6-磷酸果糖激酶的可检测水平。所有菌株都含有葡萄糖-6-磷酸脱氢酶和果糖二磷酸醛缩酶。酶研究的结果支持这样的论点,即磷酸戊糖和 Entner-Doudoroff 途径在嗜酸异养菌中起葡萄糖分解代谢的作用,而 Embden-Meyerhof-Parnas 途径显然不存在。