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Direct immobilization of a recombinant invertase to Avicel by E. coli overexpression of a fusion protein containing the extracellular invertase from Zymomonas mobilis and the carbohydrate-binding domain CBDCex from Cellulomonas fimi

Paper ID Volume ID Publish Year Pages File Format Full-Text
18228 42714 2006 5 PDF Available
Title
Direct immobilization of a recombinant invertase to Avicel by E. coli overexpression of a fusion protein containing the extracellular invertase from Zymomonas mobilis and the carbohydrate-binding domain CBDCex from Cellulomonas fimi
Abstract

The Zymomonas mobilis gene invB, encoding an extracellular invertase, was translationally fused to the cellulose binding domain Cex (CBDCex) from Cellulomonas fimi and expressed in Escherichia coli. The fusion protein obtained, termed INVB-CBD, was purified and immobilized to Avicel (crystalline cellulose) in a single step. INVB-CBD showed an invertase activity of 20.8 IU/mL and was able to bind Avicel at a ratio of 5.3 mg of protein per gram of Avicel in 5 min. Both free and immobilized INVB-CBD showed the same optimal pH (5.5) than native INVB. However, an optimal temperature shift of −10 °C was observed for both free and immobilized INVB-CBD (45 °C) compared with the native INVB (55 °C). The Km value for the free and immobilized INVB-CBD was 193 and 81 mM of sucrose, respectively; whereas, the Km value for the native INVB was 208 mM of sucrose. Thus, suggesting an improvement in the substrate affinity of INVB after the addition of the CBDCex tag. Additionally, a decrease in the Vm value was found for the free and immobilized INVB-CBD compared with the native INVB.

Keywords
Zymomonas mobilis; Invertase; Fusion protein; CBD; Enzyme immobilization
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Direct immobilization of a recombinant invertase to Avicel by E. coli overexpression of a fusion protein containing the extracellular invertase from Zymomonas mobilis and the carbohydrate-binding domain CBDCex from Cellulomonas fimi
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 40, Issue 1, 6 December 2006, Pages 172–176
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us