Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris

Zhenya Chen; Ye Li; Qipeng Yuan

doi:10.1016/j.ijbiomac.2014.07.040

Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris

Zhenya Chen, Ye Li^*, Qipeng Yuan

^*Corresponding author for this work

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

23 Citations (Scopus)

Abstract

Chondroitinase ABC I (ChSase ABC I) which can degrade chondroitin sulfate (CS) and other glycosaminoglycan to oligosaccharide or unsaturated disaccharide, was fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3) (E. coli BL21(DE3)) and purified for the first time in this study. The result showed that the productivity of recombinant MBP-ChSase ABC I was 3180IU/(L fermentation liquor) with CS A as substrate, and the productivity might be the highest level when compared to the reported ones. The specific activity of recombinant MBP-ChSase ABC I was 76IU/(mg protein) after purification. The V_max, K_m and k_cat were 18.7±0.3μmol/Ls, 73.1±4.1μmol/L and 586.7±10.8s^-1, respectively. Enzyme activity of the purified enzyme remained about 78% after 210min when the enzyme incubated at 30°C. This study introduces a rapid method for highly expressing ChSase ABC I, and the method could be adopted in the process of industrial production. Furthermore the investigation of thermostability might lead to an important guide in clinical treatment.

Original language	English
Pages (from-to)	6-10
Number of pages	5
Journal	International Journal of Biological Macromolecules
Volume	72
DOIs	https://doi.org/10.1016/j.ijbiomac.2014.07.040
Publication status	Published - Jan 2015
Externally published	Yes

Keywords

Characterization
Expression
MBP-chondrotinase ABC I
Productivity
Thermostability

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10.1016/j.ijbiomac.2014.07.040

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title = "Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris",

abstract = "Chondroitinase ABC I (ChSase ABC I) which can degrade chondroitin sulfate (CS) and other glycosaminoglycan to oligosaccharide or unsaturated disaccharide, was fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3) (E. coli BL21(DE3)) and purified for the first time in this study. The result showed that the productivity of recombinant MBP-ChSase ABC I was 3180IU/(L fermentation liquor) with CS A as substrate, and the productivity might be the highest level when compared to the reported ones. The specific activity of recombinant MBP-ChSase ABC I was 76IU/(mg protein) after purification. The Vmax, Km and kcat were 18.7±0.3μmol/Ls, 73.1±4.1μmol/L and 586.7±10.8s-1, respectively. Enzyme activity of the purified enzyme remained about 78% after 210min when the enzyme incubated at 30°C. This study introduces a rapid method for highly expressing ChSase ABC I, and the method could be adopted in the process of industrial production. Furthermore the investigation of thermostability might lead to an important guide in clinical treatment.",

keywords = "Characterization, Expression, MBP-chondrotinase ABC I, Productivity, Thermostability",

author = "Zhenya Chen and Ye Li and Qipeng Yuan",

year = "2015",

month = jan,

doi = "10.1016/j.ijbiomac.2014.07.040",

language = "English",

volume = "72",

pages = "6--10",

journal = "International Journal of Biological Macromolecules",

issn = "0141-8130",

publisher = "Elsevier B.V.",

}

TY - JOUR

T1 - Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris

AU - Chen, Zhenya

AU - Li, Ye

AU - Yuan, Qipeng

PY - 2015/1

Y1 - 2015/1

N2 - Chondroitinase ABC I (ChSase ABC I) which can degrade chondroitin sulfate (CS) and other glycosaminoglycan to oligosaccharide or unsaturated disaccharide, was fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3) (E. coli BL21(DE3)) and purified for the first time in this study. The result showed that the productivity of recombinant MBP-ChSase ABC I was 3180IU/(L fermentation liquor) with CS A as substrate, and the productivity might be the highest level when compared to the reported ones. The specific activity of recombinant MBP-ChSase ABC I was 76IU/(mg protein) after purification. The Vmax, Km and kcat were 18.7±0.3μmol/Ls, 73.1±4.1μmol/L and 586.7±10.8s-1, respectively. Enzyme activity of the purified enzyme remained about 78% after 210min when the enzyme incubated at 30°C. This study introduces a rapid method for highly expressing ChSase ABC I, and the method could be adopted in the process of industrial production. Furthermore the investigation of thermostability might lead to an important guide in clinical treatment.

AB - Chondroitinase ABC I (ChSase ABC I) which can degrade chondroitin sulfate (CS) and other glycosaminoglycan to oligosaccharide or unsaturated disaccharide, was fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3) (E. coli BL21(DE3)) and purified for the first time in this study. The result showed that the productivity of recombinant MBP-ChSase ABC I was 3180IU/(L fermentation liquor) with CS A as substrate, and the productivity might be the highest level when compared to the reported ones. The specific activity of recombinant MBP-ChSase ABC I was 76IU/(mg protein) after purification. The Vmax, Km and kcat were 18.7±0.3μmol/Ls, 73.1±4.1μmol/L and 586.7±10.8s-1, respectively. Enzyme activity of the purified enzyme remained about 78% after 210min when the enzyme incubated at 30°C. This study introduces a rapid method for highly expressing ChSase ABC I, and the method could be adopted in the process of industrial production. Furthermore the investigation of thermostability might lead to an important guide in clinical treatment.

KW - Characterization

KW - Expression

KW - MBP-chondrotinase ABC I

KW - Productivity

KW - Thermostability

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U2 - 10.1016/j.ijbiomac.2014.07.040

DO - 10.1016/j.ijbiomac.2014.07.040

M3 - Article

C2 - 25077839

AN - SCOPUS:84906336174

SN - 0141-8130

VL - 72

SP - 6

EP - 10

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

ER -

Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris

Abstract

Keywords

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