Purification of recombinant hyaluronan lyase of Streptococcus pyogenes bacteriophage H4489A expressed in Escherichia coli and its application for the specific determination of hyaluronan concentration

Hyaluronan (HA) lyase of Streptococcus pyogenes bacteriophage was expressed in Escherichia coli and purified to homogeneity by immobilized metal affinity chromatography (IMAC). Unlike most bacterial HA lyases, the phage enzyme specifically cleaved HA to unsaturated oligosaccharides which has an opti...

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Veröffentlicht in:Carbohydrate polymers 2006-07, Vol.65 (2), p.159-164
Hauptverfasser: Yang, Pei-Fen, Lee, Cheng-Kang
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Lee, Cheng-Kang
description Hyaluronan (HA) lyase of Streptococcus pyogenes bacteriophage was expressed in Escherichia coli and purified to homogeneity by immobilized metal affinity chromatography (IMAC). Unlike most bacterial HA lyases, the phage enzyme specifically cleaved HA to unsaturated oligosaccharides which has an optimum absorption at 232 nm. The absorbance of the digestion product reached a limiting value as reaction time increased. The limiting absorbance showed linearity in the range of concentrations 0.05–0.5 mg/mL. Based on this fact, a specific, simple, easy to apply, low cost, and fast enough method was developed for routine determination of HA concentration of a microbial HA production process. This phage HA lyase-based limiting absorbance method has same accuracy and sensitivity as conventional carbazole method for the determination of HA produced from Streptococcus zooepidemicus cultivation.
doi_str_mv 10.1016/j.carbpol.2005.12.037
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source Elsevier ScienceDirect Journals Complete
subjects Biological and medical sciences
Biotechnology
Carbazole method
Escherichia coli
Fundamental and applied biological sciences. Psychology
Hyaluronan
Hyaluronan determination
Limiting absorbance
Phage ha
Phage hyaluronan lyase
Streptococcus pyogenes
title Purification of recombinant hyaluronan lyase of Streptococcus pyogenes bacteriophage H4489A expressed in Escherichia coli and its application for the specific determination of hyaluronan concentration
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