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 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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. |
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ISSN: | 0144-8617 1879-1344 |
DOI: | 10.1016/j.carbpol.2005.12.037 |