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Applied Microbiology and Biotechnology

, Volume 67, Issue 6, pp 778–788 | Cite as

Purification and characterization of a biodegradable plastic-degrading enzyme from Aspergillus oryzae

  • Hiroshi Maeda
  • Youhei Yamagata
  • Keietsu Abe
  • Fumihiko Hasegawa
  • Masayuki Machida
  • Ryoji Ishioka
  • Katsuya Gomi
  • Tasuku Nakajima
Biotechnologically Relevant Enzymes and Proteins

Abstract

We used biodegradable plastics as fermentation substrates for the filamentous fungus Aspergillus oryzae. This fungus could grow under culture conditions that contained emulsified poly-(butylene succinate) (PBS) and emulsified poly-(butylene succinate-co-adipate) (PBSA) as the sole carbon source, and could digest PBS and PBSA, as indicated by clearing of the culture supernatant. We purified the PBS-degrading enzyme from the culture supernatant, and its molecular mass was determined as 21.6 kDa. The enzyme was identified as cutinase based on internal amino acid sequences. Specific activities against PBS, PBSA and poly-(lactic acid) (PLA) were determined as 0.42 U/mg, 11 U/mg and 0.067 U/mg, respectively. To obtain a better understanding of how the enzyme recognizes and hydrolyzes PBS/PBSA, we investigated the environment of the catalytic pocket, which is divided into carboxylic acid and alcohol recognition sites. The affinities for different substrates depended on the carbon chain length of the carboxylic acid in the substrate. Competitive inhibition modes were exhibited by carboxylic acids and alcohols that consisted of C4-C6 and C3-C8 chain lengths, respectively. Determination of the affinities for different chemicals indicated that the most preferred substrate for the enzyme would consist of butyric acid and n-hexanol.

Keywords

Succinic Acid Hexanoic Acid Biodegradable Plastic Inhibition Mode Butylene Succinate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work has been supported by Innovation Plaza Miyagi of JST (Japan Science and Technology Agency), and was supported in part by a grant-in-aid (Bio Design Program) from the Ministry of Agriculture, Forestry and Fisheries of Japan (BDP-03-VI-1-6)

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Hiroshi Maeda
    • 1
    • 2
    • 3
  • Youhei Yamagata
    • 2
    • 3
  • Keietsu Abe
    • 2
    • 3
  • Fumihiko Hasegawa
    • 3
  • Masayuki Machida
    • 4
  • Ryoji Ishioka
    • 5
  • Katsuya Gomi
    • 3
    • 6
  • Tasuku Nakajima
    • 2
  1. 1.Tohoku TechnoarchSendaiJapan
  2. 2.Laboratory of Molecular Enzymology, Division of Life Science, Graduate School of Agricultural ScienceTohoku UniversitySendaiJapan
  3. 3.New Industry Creation Hatchery CenterTohoku UniversitySendaiJapan
  4. 4.Gene Regulation Group, Molecular and Cell BiologyNational Institute of Advanced Industrial Science and TechnologyTsukubaJapan
  5. 5.Showa HighpolymerKanda Chuo Bldg. 20TokyoJapan
  6. 6.Laboratory of Bioindustrial Genomics, Division of Bioscience and Biotechnology for Future Bioindustries, Graduate School of Agricultural ScienceTohoku UniversitySendaiJapan

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