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Direct recovery of alcohol dehydrogenase from unclarified yeast cell homogenate by IDEBAC using an improved scheme for elution

Paper ID Volume ID Publish Year Pages File Format Full-Text
4916 258 2006 10 PDF Available
Title
Direct recovery of alcohol dehydrogenase from unclarified yeast cell homogenate by IDEBAC using an improved scheme for elution
Abstract

Procion Red HE-7B was directly immobilized onto STREAMLINE matrix to obtain a moderately substituted dye-ligand matrix in which the dye concentration was 6.72 μmol/ml adsorbent. The adsorption characteristics of alcohol dehydrogenase (ADH) onto the sticky dyed adsorbent in the absence and/or presence of yeast cells and cell debris were investigated. The Langmuir-type isotherm model was reasonably used to describe the adsorption behavior. The results showed that the adsorption characteristics and uptake rate for ADH did not change significantly even in the presence of cells and cell debris. Measurements of adsorption breakthrough curves in expanded beds have shown that the maximum dynamic binding capacity appeared to be significantly affected by the aspect ratio. The dynamic binding capacity of the dyed adsorbent was found to be 50.75 U/ml as the aspect ratio was up to 16. Optimal conditions for elution of the adsorbed ADH were determined in small scale packed bed experiments conducted with clarified yeast cell homogenates. The one-step elution was carried out by injecting a pulse of NAD+ (15 mM, one settled bed volume) with 0.3 M NaCl as a carrier, resulting in the adsorbed ADH eluting completely from the sticky dyed adsorbent. This case study on the adsorption/desorption characteristics of the sticky dyed adsorbent for ADH showed that this enzyme could be recovered with a purification factor of 4.6 and yield of 89% by immobilized dye-ligand expanded bed affinity chromatography (IDEBAC) in a single step.

Keywords
ADH, alcohol dehydrogenase; DF, dilution factor; EBA, expanded bed adsorption; EBHIC, expanded bed hydrophobic interaction chromatography; G6PDH, glucose-6-phosphate dehydrogenase; IDA, iminodiacetic acid; IDEBAC, immobilized dye-ligand expanded bed affin
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Direct recovery of alcohol dehydrogenase from unclarified yeast cell homogenate by IDEBAC using an improved scheme for elution
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 30, Issue 1, 1 May 2006, Pages 1–10
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