Ohhata Naoko, Yoshida Nobuyuki, Egami Hiroshi, Katsuragi Tohoru, Tani Yoshiki, Takagi Hiroshi
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
J Bacteriol. 2007 Oct;189(19):6824-31. doi: 10.1128/JB.00872-07. Epub 2007 Aug 3.
Rhodococcus erythropolis N9T-4, which was isolated from crude oil, showed extremely oligotrophic growth and formed its colonies on a minimal salt medium solidified using agar or silica gel without any additional carbon source. N9T-4 did not grow under CO(2)-limiting conditions but could grow on a medium containing NaHCO(3) under the same conditions, suggesting that the oligotrophic growth of N9T-4 depends on CO(2). Proteomic analysis of N9T-4 revealed that two proteins, with molecular masses of 45 and 55 kDa, were highly induced under the oligotrophic conditions. The primary structures of these proteins exhibited striking similarities to those of methanol: N,N'-dimethyl-4-nitrosoaniline oxidoreductase and an aldehyde dehydrogenase from Rhodococcus sp. These enzyme activities were three times higher under oligotrophic conditions than under n-tetradecane-containing heterotrophic conditions, and gene disruption for the aldehyde dehydrogenase caused a lack of growth on the minimal salt medium. Furthermore, 3-hexulose 6-phosphate synthase and phospho-3-hexuloisomerase activities, which are key enzymes in the ribulose monophosphate pathway in methylotrophic bacteria, were detected specifically in the cell extract of oligotrophically grown N9T-4. These results suggest that CO(2) fixation involves methanol (formaldehyde) metabolism in the oligotrophic growth of R. erythropolis N9T-4.
从原油中分离得到的红平红球菌N9T-4表现出极低营养型生长,并且在不添加任何额外碳源的情况下,能在以琼脂或硅胶固化的基本盐培养基上形成菌落。N9T-4在二氧化碳限制条件下不能生长,但在相同条件下可在含有碳酸氢钠的培养基上生长,这表明N9T-4的极低营养型生长依赖于二氧化碳。对N9T-4的蛋白质组分析表明,有两种分子量分别为45 kDa和55 kDa的蛋白质在极低营养条件下被高度诱导。这些蛋白质的一级结构与甲醇:N,N'-二甲基-4-亚硝基苯胺氧化还原酶和红球菌属的一种醛脱氢酶的一级结构有显著相似性。这些酶活性在极低营养条件下比在含正十四烷的异养条件下高3倍,醛脱氢酶的基因破坏导致在基本盐培养基上无法生长。此外,在极低营养条件下生长的N9T-4的细胞提取物中特异性检测到了6-磷酸-3-己酮糖合酶和磷酸-3-己酮异构酶的活性,这两种酶是甲基营养细菌中磷酸戊糖途径的关键酶。这些结果表明,在红平红球菌N9T-4的极低营养型生长中,二氧化碳固定涉及甲醇(甲醛)代谢。