Prediction of optimum reaction conditions for the thermo-tolerant acetylxylan esterase from Neocallimastix patriciarum using the response surface methodology
Journal
Journal of Chemical Technology and Biotechnology
Journal Volume
85
Journal Issue
5
Pages
628-633
Date Issued
2010
Author(s)
Abstract
BACKGROUND: Xylan is the second most abundant renewable polysaccharide in nature and also represents an important industrial substrate. The complete degradation of xylan requires the combination of several types of xylanolytic enzymes, including endo-β-1,4-xylanases, β-xylosidases, and acetylxylan esterases. As a biocatalyst, xylanolytic enzymes with good thermal stability are of great interest, therefore, a thermo-tolerant acetylxylan esterase, AxeS20E, was investigated. RESULTS: The cDNAencoding the carbohydrate esterase (CE) domain of AxeS20E from Neocallimastixpatriciarum was expressed in Escherichia coli as a recombinant His6 fusion protein. The recombinant AxeS20E protein was obtained after purification by immobilized metal ion-affinity chromatography. Response surface modeling (RSM) combined with central composite design (CCD) and regression analysis were then employed for the planned statistical optimization of the acetylxylan esterase activities of AxeS20E. The optimal conditions for the highest activity of AxeS20E were observed at 54.6 °C and pH 7.8. Furthermore, AxeS20E retained more than 85% of its initial activity after 120 min of heating at 80 °C. CONCLUSIONS: The results suggested that RSM combined with CCD and regression analysis were effective in determining optimized temperature and pH conditions for the enzyme activity of AxeS20E. The results also proved AxeS20E was thermotolerant andmight be a good candidate for various biotechnological applications. © 2010 Society of Chemical Industry.
Subjects
Acetylxylan esterase; Optimization; Response surface methodology; Thermo-tolerant
SDGs
Other Subjects
Biotechnological applications; Central composite designs; Esterase activities; Fusion proteins; Immobilized metals; Initial activity; Optimal conditions; Optimum reaction conditions; pH condition; Response Surface Methodology; Response surface modeling; Statistical optimization; Thermal stability; Thermotolerant; Xylanases; Xylanolytic enzymes; Affinity chromatography; Carbohydrates; Catalysts; Degradation; Digital cameras; Enzyme activity; Enzymes; Escherichia coli; Heating; Metal ions; Metal refining; Regression analysis; Surface properties; Thermogravimetric analysis; Optimization; acetylxylan esterase; complementary DNA; fungal enzyme; histone; hybrid protein; metal ion; unclassified drug; affinity chromatography; article; biocatalyst; enzyme activity; Escherichia coli; heat tolerance; Neocallimastix; Neocallimastix patriciarum; nucleotide sequence; pH; prediction; protein expression; protein purification; regression analysis; response surface method; thermostability; Escherichia coli; Neocallimastix patriciarum
Type
journal article