Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A

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

doi:10.1016/j.carbpol.2011.01.019

Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A

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

^*Corresponding author for this work

Department of Life Science

Research output: Contribution to journal › Article › peer-review

11 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 language	English
Pages (from-to)	1182-1191
Number of pages	10
Journal	Carbohydrate Polymers
Volume	84
Issue number	3
DOIs	https://doi.org/10.1016/j.carbpol.2011.01.019
Publication status	Published - 2011 Mar 17

Keywords

Characterization
Hyaluronate lyase
Kinetics
Purification
Structure

ASJC Scopus subject areas

Organic Chemistry
Polymers and Plastics
Materials Chemistry

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10.1016/j.carbpol.2011.01.019

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title = "Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A",

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.",

keywords = "Characterization, Hyaluronate lyase, Kinetics, Purification, Structure",

author = "El-Safory, {Nermeen S.} and Lee, {Guan Chiun} and Lee, {Cheng Kang}",

year = "2011",

month = mar,

day = "17",

doi = "10.1016/j.carbpol.2011.01.019",

language = "English",

volume = "84",

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journal = "Carbohydrate Polymers",

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T1 - Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A

AU - El-Safory, Nermeen S.

AU - Lee, Guan Chiun

AU - Lee, Cheng Kang

PY - 2011/3/17

Y1 - 2011/3/17

N2 - 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.

AB - 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.

KW - Characterization

KW - Hyaluronate lyase

KW - Kinetics

KW - Purification

KW - Structure

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DO - 10.1016/j.carbpol.2011.01.019

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SN - 0144-8617

VL - 84

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JO - Carbohydrate Polymers

JF - Carbohydrate Polymers

IS - 3

ER -

Characterization of hyaluronate lyase from Streptococcus pyogenes bacteriophage H4489A

Abstract

Keywords

ASJC Scopus subject areas

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