Abstract
We investigated the optimum conditions for the formation of nitrile hydratase (NHase), which acts on indole-3-acetonitrile, in Agrobacterium tumefaciens. Good inducers for enzyme formation have been found to be roughly classified into three representative types of amides such as pivalamide, crotonamide and e-caprolactam. When the strain was cultivated in the optimum culture medium containing ε-caprolactam as an inducer, in particular, the specific activity of NHase in the culture was 13 000 times higher than that without addition of amides, nitriles or acids. In this case, NHase formed accounted for 12% of the total cellular soluble protein. The purified NHase did not act on ε-caprolactam, and ε-caprolactam was not degraded during the cultivation by the strain, suggesting that ε-caprolactam seems to keep driving the NHase induction mechanism.
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References
Duran R, Nishiyama M, Horinouchi S, Beppu T (1993) Characterization of nitrile hydratase genes cloned by DNA screening from Rhodococcus erythropolis. Biosci Biotechnol Biochem 57: 1323–1328
Kobayashi M, Shimizu S (1994) Versatile nitrilases: nitrile-hydrolyzing enzymes. FEMS Microbiol Lett 120: 217–224
Kobayashi M, Nagasawa T, Yamada H (1992) Enzymatic synthesis of acrylamide: a success story not yet over. Trends Biotechnol 10: 402–408
Kobayashi M, Izui H, Nagasawa T, Yamada H (1993) Nitrilase in biosynthesis of the plant hormone indole-3-acetic acid from indole-3-acetonitrile: cloning of the Alcaligenes gene and site-directed mutagenesis of cysteine residues. Proc Natl Acad Sci USA 90: 247–251
Kobayashi M, Suzuki T, Fujita T, Masuda M, Shimizu S (1995) Occurrence of enzymes involved in biosynthesis of indole-3-acetic acid from indole-3-acetonitrile in plant-associated bacteria, Agrobacterium and Rhizobium. Proc Natl Acad Sci USA 92: 714–718
Nagasawa T, Yamada H (1989) Microbial transformations of nitriles. Trends Biotechnol 7: 153–158
Nagasawa T, Kobayashi M, Yamada H (1988) Optimum culture conditions for the production of benzonitrilase by Rhodococcus rhodochrous J1. Arch Microbiol 150: 89–94
Nagasawa T, Nakamura T, Yamada H (1990) ε-Caprolactam, a new powerful inducer for the formation of Rhodococcus rhodochrous J1 nitrilase. Arch Microbiol 155: 13–17
Sugiura T, Kuwahara J, Nagasawa T, Yamada H (1987) Nitrile hydratase: the first non-heme iron enzyme with a typical low-spin Fe(III) active center. J Am Chem Soc 109: 5848–5850
Thimann KV, Mahadevan S (1964) Nitrilase: I. Occurrence, preparation and general properties of the enzyme. Arch Biochem Biophys 105: 133–141
Thomashow MF, Hugly S, Buchholz WG, Thomashow LS (1986) Molecular basis for the auxin-independent phenotype of crown gall tumor tissues. Science 231: 616–618
Yamada H, Ryuno K, Nagasawa T, Enomoto K, Watanabe I (1986) Optimum culture conditions for production by Pseudomonas chlororaphis B23 of nitrile hydratase. Agric Biol Chem 50: 2859–2865
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Kobayashi, M., Fujita, T. & Shimizu, S. Hyperinduction of nitrile hydratase acting on indole-3-acetonitrile in Agrobacterium tumefaciens . Appl Microbiol Biotechnol 45, 176–181 (1996). https://doi.org/10.1007/s002530050667
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DOI: https://doi.org/10.1007/s002530050667