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Immobilization of l-lactate dehydrogenase on magnetic nanoclusters for chiral synthesis of pharmaceutical compounds

Paper ID Volume ID Publish Year Pages File Format Full-Text
4048 206 2009 9 PDF Available
Title
Immobilization of l-lactate dehydrogenase on magnetic nanoclusters for chiral synthesis of pharmaceutical compounds
Abstract

In the present work, we study the covalent immobilization of l-lactate dehydrogenase (LDH) on silica-coated magnetic nanoclusters (SiMag). Immobilization of LDH on magnetic nanoclusters provides several advantages. First, it can ease the enzyme recovery through magnetic separation, rendering the immobilized LDH applicable for batch reactor which configuration can minimize the product inhibition. Second, the immobilization resulted in the increase of product inhibition constants of LDH which can further minimize the product inhibition. A 5-fold and a 1.6-fold increase of apparent competitive product inhibition constant of NAD+ (Kiq) and of lactate (KiqKp/Kq) were observed, respectively. Third, the immobilized LDH showed improvement on thermal stability, with 5–7-fold reduced deactivation rate constant in second phase (A) of biphasic deactivation behavior, but with no significant difference in the first phase (B). Present study also develops a method to improve the immobilization process by understanding the surface interaction involved during the covalent immobilization of enzyme on magnetic nanoclusters. We found that the addition of sodium chloride during the immobilization could improve the relative activity of immobilized LDH from 50% to about 75%. This was due to the possibility of hydrophobic interaction in controlling a better LDH orientation on particle surface, and thus avoiding any severe deactivation. Protein representation using Jmol software also confirmed this possibility.

Keywords
Dehydrogenase; Immobilized enzyme; Magnetic nanoclusters; Oriented immobilization; Stability
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Immobilization of l-lactate dehydrogenase on magnetic nanoclusters for chiral synthesis of pharmaceutical compounds
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 48, Issue 1, 15 December 2009, Pages 13–21
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
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