fulltext.study @t Gmail

Recovery of recombinant β-glucosidase by expanded bed adsorption from Pichia pastoris high-cell-density culture broth

Paper ID Volume ID Publish Year Pages File Format Full-Text
25580 43582 2006 13 PDF Available
Title
Recovery of recombinant β-glucosidase by expanded bed adsorption from Pichia pastoris high-cell-density culture broth
Abstract

Methanol limited fed-batch cultivation was applied for production of a plant derived β-glucosidase by Pichia pastoris. The β-glucosidase was recovered by expanded bed adsorption chromatography applied to the whole culture broth. The new Streamline Direct HST1 adsorbent was compared with Streamline SP. Higher bead density made it possible to operate at two times higher feedstock concentration and at two times higher flow velocity. The higher binding capacity in the conductivity range 0–48 mS cm−1 of Streamline Direct HST1 might be caused by the more complex interaction of multi-modal ligand in Streamline Direct HST1 compared to the single sulphonyl group in Streamline SP. Harsher elution condition had to be applied for dissociation of β-glucosidase from Streamline Direct HST1 due to stronger binding interaction. The 5% dynamic binding capacity was 160 times higher for Streamline Direct HST1 compared to Streamline SP. The yield of β-glucosidase on Streamline Direct HST1 (74%) was significantly higher than on Streamline SP (48%). Furthermore, β-glucosidase was purified with a factor of 4.1 and concentrated with a factor of 17 on Streamline Direct HST1 while corresponding parameters were half of these values for Streamline SP. Thus, for all investigated parameters Streamline Direct HST1 was a more suitable adsorbent for recovery of recombinant β-glucosidase from unclarified P. pastoris high-cell-density cultivation broth.

Keywords
AOX, enzyme alcohol oxidase; aox, alcohol oxidase gene; EBA, expanded bed adsorption; PI, propidium iodideβ-Glucosidase; Pichia pastoris; Methanol limited fed-batch; Expanded bed adsorption; Multi-modal ligand
First Page Preview
Recovery of recombinant β-glucosidase by expanded bed adsorption from Pichia pastoris high-cell-density culture broth
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 122, Issue 1, 9 March 2006, Pages 86–98
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering