Microbial Fixation of CO_2 to Organic Compounds and Its Application to the Production of useful materials

当红球红球菌An-13在苯胺上生长时，在培养液中积累了一种荧光物质。得到晶体，鉴定为邻氨基苯甲酸(Ana)。在与生长在苯胺上的细菌的休眠细胞孵育后，也可以从苯胺中产生ANA。加热的细胞失去了产生它的活性，而苯胺是产生它的必需品。碳酸氢钠促进了ANA的产生，当在培养混合物中加入^&lt；14&gt；C-碳酸氢钠时，会生成^&lt；14&gt；C-Ana。静息细胞产生ANA的最适pH为7.0~7.5。这些结果表明，红假单胞菌An-13的微生物活性催化了苯胺合成ANA。对40株细菌在有无苯胺的情况下进行了生产研究。所有苯胺同化菌的休眠细胞都比其他苯胺同化菌产生更多的ANA。几株非苯胺同化S…的休眠细胞更多的火车在没有苯胺的情况下产生了ANA。然而，这些菌株的产量远远低于红球菌菌株。脂肪单羧酸盐、三磷酸腺苷、生物素和辅酶A可促进苯胺同化红曲霉An-13细胞合成ANA，而邻苯二酚类似物N-乙基马来酰亚胺和碘乙酸酯则抑制其合成ANA。另一方面，利用非苯胺类同化假单胞菌进行发酵生产。AN21受葡萄糖、甘露糖和一些氨基酸的抑制，红球藻AN-13细胞的无细胞提取液保持了从苯胺产生ANA的活性。这些提取物中的催化因子(S)经硫酸链霉素和硫酸铵分级沉淀和Tyopearl HW55s凝胶过滤后得到部分纯化。对这一催化因子(S)的进一步纯化过程正在进行中。分离到一株通过歧解途径代谢苯胺的细菌FK-8-2，经鉴定为假单胞菌。假单胞菌属FK-8-2细胞产生大量邻苯二酚2，3-双加氧酶。对该酶进行了纯化，得到了均一酶。纯化后的酶与恶臭假单胞菌Mt-2在化学性质和催化性质上有所不同。还分离到了另一株在细胞内产生邻苯二酚1，2双加氧同工酶的苯胺同化细菌。这些菌株为研究苯胺法生产苯丙氨酸提供了有用的材料。较少

When Rhodococcus erythropolis AN-13 grew on aniline, a fluorescent substance accumulated in the cultural fluid. It was obtained as crystals and identified as anthranilic acid (AnA). AnA was also produced from aniline following incubation with resting cells of the bacterium grown on aniline. Heated cells lost the activity to produce it, and aniline was essential for its production. The production of AnA was promoted by sodium bicapbonate ; when ^<14>C-sodium bicarbonate was added to the incubation mixture, ^<14>C-AnA formed. the optimal pH for AnA production by the resting cells was 7.0 to 7.5. These results suggest that microbial activities of R. erythropolis AN-13 catalyzed the formation of AnA from aniline.The production was investigated for 40 bacterial strains in the presence and absence of aniline. Resting cells of all aniline-assimilating R. erythropolis strains produced more AnA than those of other anilineassimilating bacteria. Resting cells of several non-aniline-assimilating s … More trains produced AnA in the absence of aniline. However, its production by these strains was much lower than that by the Rhodococcus strains. The production of AnA by cells of aniline-assimilating R. erythropolis AN-13 was promoted by aliphatic monocarboxylates, ATP, biotin and coenzyme A, and repressed by catechol analogues, N-ethylmaleimide and iodoacetate. on the other hand, its production by non-aniline-assimilating PseudoNonas sp. AN21 was repressed by glucose, mannose and some amino acids.Cell-free extracts prepared from R. erythropolis AN-13 cells retained the activity capable of producing AnA from aniline. The catalytic factor(s) in these extracts was partially purified by fractionation with streptomycin sulfate and anunonium sulfate and gel filtration on a column of Tyopearl HW55s. Further purification procedures of this catalytic factor(s) are in progress.A bacterial strain, FK-8-2, that metabolized aniline through a metacleavage pathway was isolated and identified as Pseudomonas. Pseudomonas sp. FK-8-2 produced much catechol 2, 3-dioxygenase in its cells. This enzyme was purified to homogeneity. The purified enzyme differed in chemical and catalytic properties from that obtained from Pseudomonas putida mt-2 growing on benzoate. Another aniline-assimilating bacteria producing isozymes of catechol 1, 2dioxygense in their cells were also isolated. These isolates will be useful materials for investigating on the bacterial production of AnA from aniline. Less