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Improvement of recombinant endoglucanase produced in Pichia pastoris KM71 through the use of synthetic medium for inoculum and pH control of proteolysis

Paper ID Volume ID Publish Year Pages File Format Full-Text
20620 43183 2013 6 PDF Available
Title
Improvement of recombinant endoglucanase produced in Pichia pastoris KM71 through the use of synthetic medium for inoculum and pH control of proteolysis
Abstract

The long lag time in basal salts medium (BSM) and an occurrence of proteolysis are major problems for recombinant protein production in Pichia pastoris KM71. In this study, optimal conditions were explored for fed-batch cultivation of recombinant fungal endoglucanase in P. pastoris KM71. It was found that lag and process times were much reduced when the synthetic FM22 medium was used for the inoculum compared with enriched buffered glycerol complex (BMGY) medium. The highest endoglucanase activity was obtained at 30°C which was more than 10 fold higher than that produced from shake flask. At 30°C, the specific endoglucanase activity was dependent on culture pH and a higher specific activity was observed at pH 5.0 than at pH 6.0. The higher activity was likely due to lower rate of proteolysis, since a truncated protein species was apparent at pH 6.0, but not pH 5.0. Thus, production of endoglucanase at 30°C and pH 5.0 is the optimal condition suitable for economical production in large scale. The combination of using synthetic FM22 medium for inoculum and proteolysis control by growth at lower pH could be applied for production of other recombinant proteins in P. pastoris.

Keywords
Inoculum development; Lag time; Pichia pastoris; Endoglucanase; Proteolysis
First Page Preview
Improvement of recombinant endoglucanase produced in Pichia pastoris KM71 through the use of synthetic medium for inoculum and pH control of proteolysis
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 116, Issue 2, August 2013, Pages 193–198
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering