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Isolation of a bacterium that degrades urethane compounds and characterization of its urethane hydrolase

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Abstract

A bacterium which degrades urethane compounds was isolated and identified as Rhodococcus equi strain TB-60. Strain TB-60 degraded toluene-2,4-dicarbamic acid dibutyl ester (TDCB) and accumulated toluene diamine as the degradation product. The enzyme which cleaves urethane bond in TDCB was strongly induced by acetanilide. The purified enzyme (urethane hydrolase) was found to be homogeneous on sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The molecular weight was estimated to be 55 kDa. The optimal temperature and pH were 45°C and 5.5, respectively. The enzyme hydrolyzed aliphatic urethane compound as well as aromatic ones. The activity was inhibited by HgCl2, p-chrolomercuribenzoic acid, and phenylmethylsulfonyl fluoride, suggesting that cysteine and/or serine residues play an important role in the activity. The enzyme catalyzed the hydrolysis of anilides, amides, and esters as well as TDCB. It was characterized as a novel amidase/esterase, differing in some properties from other known amidases/esterases.

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Abbreviations

EPC:

Ethyl N-phenylcarbamate

HDCB:

Hexamethylene dicarbamic acid dibutyl ester

HDA:

Hexamethylene diamine

HFBA:

Heptafluorobutyric acid anhydride

MDCB:

Methylene bisphenyl dicarbamic acid dibutyl ester

PMSF:

Phenylmethylsulfonyl fluoride

PUR:

Polyurethane

TDA:

Toluene diamine

TDI:

Tolylene diisocyanate

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Acknowledgement

We thank Mr. Daisuke Nishiguchi of Mitsui Takeda Chemicals Co., Ltd. for supplying urethane compounds.

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Authors and Affiliations

  1. SORST, Japan Science and Technology Agency, 4-1-8 Honcho Kawaguchi, Saitama, Japan

    Yukie Akutsu-Shigeno, Kieko Toyoshima & Toshiaki Nakajima-Kambe

  2. Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan

    Yukie Akutsu-Shigeno, Yusuke Adachi, Chise Yamada, Nobuhiko Nomura, Hiroo Uchiyama & Toshiaki Nakajima-Kambe

  3. Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki, 305-8572, Japan

    Toshiaki Nakajima-Kambe

Authors
  1. Yukie Akutsu-Shigeno
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  2. Yusuke Adachi
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  3. Chise Yamada
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  4. Kieko Toyoshima
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  5. Nobuhiko Nomura
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  6. Hiroo Uchiyama
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  7. Toshiaki Nakajima-Kambe
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Correspondence to Toshiaki Nakajima-Kambe.

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Akutsu-Shigeno, Y., Adachi, Y., Yamada, C. et al. Isolation of a bacterium that degrades urethane compounds and characterization of its urethane hydrolase. Appl Microbiol Biotechnol 70, 422–429 (2006). https://doi.org/10.1007/s00253-005-0071-1

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  • DOI: https://doi.org/10.1007/s00253-005-0071-1

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