caa a22 4500 605504830 CHVBK 20210128100618.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6812-x doi (NATIONALLICENCE)springer-10.1007/s00253-015-6812-x Imidase catalyzing desymmetric imide hydrolysis forming optically active 3-substituted glutaric acid monoamides for the synthesis of gamma-aminobutyric acid (GABA) analogs [Elektronische Daten] [Masutoshi Nojiri, Makoto Hibi, Hiroaki Shizawa, Nobuyuki Horinouchi, Yoshihiko Yasohara, Satomi Takahashi, Jun Ogawa] The recent use of optically active 3-substituted gamma-aminobutyric acid (GABA) analogs in human therapeutics has identified a need for an efficient, stereoselective method of their synthesis. Here, bacterial strains were screened for enzymes capable of stereospecific hydrolysis of 3-substituted glutarimides to generate (R)-3-substituted glutaric acid monoamides. The bacteria Alcaligenes faecalis NBRC13111 and Burkholderia phytofirmans DSM17436 were discovered to hydrolyze 3-(4-chlorophenyl) glutarimide (CGI) to (R)-3-(4-chlorophenyl) glutaric acid monoamide (CGM) with 98.1% enantiomeric excess (e.e.) and 97.5% e.e., respectively. B. phytofirmans DSM17436 could also hydrolyze 3-isobutyl glutarimide (IBI) to produce (R)-3-isobutyl glutaric acid monoamide (IBM) with 94.9% e.e. BpIH, an imidase, was purified from B. phytofirmans DSM17436 and found to generate (R)-CGM from CGI with specific activity of 0.95U/mg. The amino acid sequence of BpIH had a 75% sequence identity to that of allantoinase from A. faecalis NBRC13111 (AfIH). The purified recombinant BpIH and AfIH catalyzed (R)-selective hydrolysis of CGI and IBI. In addition, a preliminary investigation of the enzymatic properties of BpIH and AfIH revealed that both enzymes were stable in the range of pH 6-10, with an optimal pH of 9.0, stable at temperatures below 40°C, and were not metalloproteins. These results indicate that the use of this class of hydrolase to generate optically active 3-substituted glutaric acid monoamide could simplify the production of specific chiral GABA analogs for drug therapeutics. Springer-Verlag Berlin Heidelberg, 2015 Imidase nationallicence 3-substituted glutarimide nationallicence Desymmetrization nationallicence Bacterial screening nationallicence GABA synthesis nationallicence Stereoselective hydrolysis nationallicence Nojiri Masutoshi Biotechnology Development Laboratories, Kaneka Corporation, 1-8 Miyamae, 676-8688, Takasago, Hyogo, Japan aut Hibi Makoto Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut Shizawa Hiroaki Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut Horinouchi Nobuyuki Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut Yasohara Yoshihiko Biotechnology Development Laboratories, Kaneka Corporation, 1-8 Miyamae, 676-8688, Takasago, Hyogo, Japan aut Takahashi Satomi Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut Ogawa Jun Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 9961-9969 0175-7598 99:23<9961 2015 99 253 https://doi.org/10.1007/s00253-015-6812-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6812-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nojiri Masutoshi Biotechnology Development Laboratories, Kaneka Corporation, 1-8 Miyamae, 676-8688, Takasago, Hyogo, Japan aut NATIONALLICENCE 700 1- Hibi Makoto Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut NATIONALLICENCE 700 1- Shizawa Hiroaki Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut NATIONALLICENCE 700 1- Horinouchi Nobuyuki Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut NATIONALLICENCE 700 1- Yasohara Yoshihiko Biotechnology Development Laboratories, Kaneka Corporation, 1-8 Miyamae, 676-8688, Takasago, Hyogo, Japan aut NATIONALLICENCE 700 1- Takahashi Satomi Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut NATIONALLICENCE 700 1- Ogawa Jun Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 606-8502, Sakyo-ku, Kyoto, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 9961-9969 0175-7598 99:23<9961 2015 99 253