Robust trypsin coating on electrospun polymer nanofibers in rigorous conditions and its uses for protein digestion

Hye Kyung Ahn, Byoung Chan Kim, Seung Hyun Jun, Mun Seock Chang, Daniel Lopez-Ferrer, Richard D. Smith, Man Bock Gu, Sang Won Lee, Beom Soo Kim, Jungbae Kim

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

An efficient protein digestion in proteomic analysis requires the stabilization of proteases such as trypsin. In the present work, trypsin was stabilized in the form of enzyme coating on electrospun polymer nanofibers (EC-TR), which crosslinks additional trypsin molecules onto covalently attached trypsin (CA-TR). EC-TR showed better stability than CA-TR in rigorous conditions, such as at high temperatures of 40 and 50°C, in the presence of organic co-solvents, and at various pH's. For example, the half-lives of CA-TR and EC-TR were 1.42 and 231 h at 40°C, respectively. The improved stability of EC-TR can be explained by covalent linkages on the surface of trypsin molecules, which effectively inhibits the denaturation, autolysis, and leaching of trypsin. The protein digestion was performed at 40°C by using both CA-TR and EC-TR in digesting a model protein, enolase. EC-TR showed better performance and stability than CA-TR by maintaining good performance of enolase digestion under recycled uses for a period of 1 week. In the same condition, CA-TR showed poor performance from the beginning and could not be used for digestion at all after a few usages. The enzyme coating approach is anticipated to be successfully employed not only for protein digestion in proteomic analysis but also for various other fields where the poor enzyme stability presently hampers the practical applications of enzymes.

Original languageEnglish
Pages (from-to)917-923
Number of pages7
JournalBiotechnology and Bioengineering
Volume107
Issue number6
DOIs
Publication statusPublished - 2010 Dec 15

Keywords

  • Electrospun nanofibers
  • Enzyme coating
  • Protein digestion
  • Trypsin

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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  • Cite this

    Ahn, H. K., Kim, B. C., Jun, S. H., Chang, M. S., Lopez-Ferrer, D., Smith, R. D., Gu, M. B., Lee, S. W., Kim, B. S., & Kim, J. (2010). Robust trypsin coating on electrospun polymer nanofibers in rigorous conditions and its uses for protein digestion. Biotechnology and Bioengineering, 107(6), 917-923. https://doi.org/10.1002/bit.22895