fulltext.study @t Gmail

Improving the production of human interferon gamma (hIFN-γ) in Pichia pastoris cell factory: An approach of cell level

Paper ID Volume ID Publish Year Pages File Format Full-Text
34177 45008 2016 10 PDF Available
Title
Improving the production of human interferon gamma (hIFN-γ) in Pichia pastoris cell factory: An approach of cell level
Abstract

•A cell and process level strategies was carried out for enhancing hIFN-γ production.•PDI enhanced the expression of rhIFN-γ by 0.53 mg/L and 0.03 mg/mL after purification.•Codon optimization resulted about 1.8 mg/L of rhIFN-γ and 0.12 mg/mL after purification.•Process parameter optimization enhanced rhIFN-γ production up to 2.5 mg/L.

Human interferon gamma (hIFN-γ) has an antiviral, immunoregulatory and anti-tumor properties. It is majorly used for the treatment of chronic granulomatous disease and severe malignant osteopetrosis. In the present investigation, cell level strategies were applied to address the expression machinery related problems, which are bottle neck for protein expression in Pichia pastoris. The gene encoding hIFN-γ was successfully cloned and expressed in P. pastoris (GS115) under the control of AOX promoter. About 200 μg/L of recombinant human interferon gamma (rhIFN-γ) was expressed after 1% methanol induction. The expression of rhIFN-γ was further enhanced by 2.67 fold by co-expressing the gene encoding protein disulphide isomerase (PDI) along with hIFN-γ. Furthermore, the effect of codon bias was examined by cloning codon optimized hIFN-γ in P. pastoris (GS115) that resulted in about 9 fold enhancement of rhIFN-γ production. Optimization of process parameters viz., temperature, pH, methanol concentration, inoculum size and agitation rate enhanced the production of rhIFN-γ by 12.5 fold. The purification with IMAC (Immobilized metal ion affinity chromatography) showed about 56.5, 63.83 and 80% purity of rhIFN-γ from GS-hIFN-γ, GS-hIFN-γ-PDI and GS-hIFN-γopt strains, respectively.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide

Keywords
Human interferon gamma (hIFN-γ); Pichia pastoris; Protein disulfide isomerase; Codon optimization; Parameter optimization
First Page Preview
Improving the production of human interferon gamma (hIFN-γ) in Pichia pastoris cell factory: An approach of cell level
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
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 51, Issue 6, June 2016, Pages 709–718
Authors
, , ,
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