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Characterization of an indican-hydrolyzing enzyme from Sinorhizobium meliloti

Paper ID Volume ID Publish Year Pages File Format Full-Text
35608 45098 2010 5 PDF Available
Title
Characterization of an indican-hydrolyzing enzyme from Sinorhizobium meliloti
Abstract

A novel β-glucosidase capable of hydrolyzing indican to indigo was mined and isolated from Sinorhizobium meliloti using a systematic approach. The corresponding gene was amplified by PCR and overexpressed in the soluble fraction as an MBP fusion protein. The resulting enzyme easily purified to apparent homogeneity via a consecutive step in the affinity column. The recombinant enzyme was determined to be a monomer with a calculated molecular mass of 52 kDa and showed the maximum activity for indican at pH 7.0 and 45 °C. The kinetic parameters for indican, KM and Vmax, were determined to be 0.97 mM and 355.6 μM/min/mg protein, respectively, at pH 7.0 and 35 °C. Additionally, this enzyme hydrolyzed both the β-(1-4)- and β-(1-6)-glucosidic bonds and revealed a minor activity against α-d-glucosides. Furthermore, the enzyme was severely inhibited by DTT, indicating a possibility that the oxidation of amino acids could play a crucial role in the activity of the enzyme.

Keywords
Indican; Indigo blue; β-Glucosidase; Sinorhizobium meliloti; Zymogram
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Characterization of an indican-hydrolyzing enzyme from Sinorhizobium meliloti
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 45, Issue 6, June 2010, Pages 892–896
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