Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A

Nermeen S. El-Safory, Guan Chiun Lee, Cheng Kang Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Hyaluronate (HA) lyase of Streptococcus pyogenes bacteriophage H4489A, was expressed in Escherichia coli, purified, and characterized. The purified homogeneous preparation of HA lyase had a molecular mass of 40 kDa. The optimum enzymatic activity was achieved at pH ∼ 5.5 and 37 °C, and the enzyme was stable at pH profile from 4 to 7 and temperature range from 25 to 45 °C. The enzymatic activity was vaguely enhanced by Mg2+, slightly inhibited by Ca2+, triton X-100, and Tween 80, strongly inhibited by Zn2+, and completely inhibited by Cu2+, Ni2+, Co2+ and sodium dodecyl sulfate. Kinetic measurements give Michaelis constant of 0.44 mg/ml, maximal velocity of 0.20 μmol ml-1 min-1, and showed that bacteriophage HA lyase degraded the HA efficiently. Light scattering dynamic measurements determined the denaturation temperate of HA lyase of about 46 °C. Circular dichromism and UV-visible absorption spectroscopy estimated the changes in secondary structure of native and denatureated HA lyase.

Original languageEnglish
Pages (from-to)1182-1191
Number of pages10
JournalCarbohydrate Polymers
Volume84
Issue number3
DOIs
Publication statusPublished - 2011 Mar 17

Fingerprint

hyaluronate lyase
Bacteriophages
Denaturation
Sodium dodecyl sulfate
Molecular mass
Dynamic light scattering
Absorption spectroscopy
Escherichia coli
Enzymes
Kinetics
Polysorbates
Octoxynol
Sodium Dodecyl Sulfate
Temperature

Keywords

  • Characterization
  • Hyaluronate lyase
  • Kinetics
  • Purification
  • Structure

ASJC Scopus subject areas

  • Organic Chemistry
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A. / El-Safory, Nermeen S.; Lee, Guan Chiun; Lee, Cheng Kang.

In: Carbohydrate Polymers, Vol. 84, No. 3, 17.03.2011, p. 1182-1191.

Research output: Contribution to journalArticle

El-Safory, Nermeen S. ; Lee, Guan Chiun ; Lee, Cheng Kang. / Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A. In: Carbohydrate Polymers. 2011 ; Vol. 84, No. 3. pp. 1182-1191.
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