The transformation of indole by Pseudomonas putida SH1

• Main Authors: Yi-Tsen Chiang, 蔣依岑
• Other Authors: Shir-Ly Huang
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/12665641774671427674

碩士 === 國立中央大學 === 生命科學研究所 === 87 === Pseudomonas putida SH1 was capable to utilize naphthalene and phenol as sole carbon source and energy to grow. This study was focused on the transformation of indole by P. putida SH1 grown from naphthalene or phenol. The major intermediates and products from indole tranaformation were characterized and the pathways were proposed. The optimal concentration of naphthalene in MSB (minimal salts basal medium) for P. putida SH1 to grow was 0.1% and that for phenol was 0.05%. According to the major products, indigo, the optimal biotransformation concentration of indole in the preseuce of 0.1% naphthalene was 2.5 mM. When the carbon source was changed to 0.05% phenol, the optimal concentration of indole was 1 mM. In order to compare the intermediates and products of indole transformation induced by naphthalene or phenol, the concentration of carbon source and indole was 0.05% and 1 mM, respectively, in both the analysis by thin layer chromatography and high pressure liquid chromatography. The products of indole transformation by P. putida SH1 grown in naphthalene were oxindole, indigo, isatin, and seven other unknown colored compounds. The indole was used up at 10 hours and the products increased up to 25 hours. The products of indole transformation by P. putida SH1 induced by phenol were 2-hydroxy indole (oxindole), indigo, isatin, and three other unknown color compounds. The varieties of the color products induced under the presence of phenol were less than those induced by naphthalene. The indole was not used up at the transformation experiment for 100 hours. The proposed enzyme induced by naphthalene for indole transformation was naphthalene dioxygenase in P. putida SH1. Indole may be oxidized by naphthalene dioxygenase to form 2,3-dihydroxy-2,3-dihydroindole, which was then spontaneously dehydrated to give hydroxy indoles. Condensation of two molecules of 3-hydroxy indole (indoxyl) followed by air oxidation or oxidation of isatin led to the production of indigo. The condensation of 2-hydroxy indole and 3-hydroxy indole yield indirubin. Isatin may be formed by the oxidiation of 2,3-dihydroxy-2,3-dihydroindole, 2-hydroxy indole, or 3-hydroxy indole or by the decomposition of indigo or indirubin. No evidence was found for the formation of other color products and they may be formed from hydroxy indoles. Indole may be oxidized by phenol hydroxylase to form hydroxy indoles in the phenol-grown SH1 cells. The formation of indigo and isatin may go through the same pathway as descirbed above.