fulltext.study @t Gmail

Cloning and expression of l-asparaginase from E. coli in eukaryotic expression system

Paper ID Volume ID Publish Year Pages File Format Full-Text
2869 140 2015 4 PDF Available
Title
Cloning and expression of l-asparaginase from E. coli in eukaryotic expression system
Abstract

•Asparaginase is an oncolytic agent which affects the growth of cancer cells.•Pichia pink expression system has high expression rate.•Cloning of Escherichia coli ansB gene in eukaryotic expression system.•Increased activity and expression level.

l-Asparaginase is an anti-cancer agent which prevents the proliferation of cancerous cells by decreasing the level of asparagine in the blood. l-asparaginase from Escherichia coli which is encoded by ansB gene is widely used because of its substrate specificity and less glutaminase activity. Here, we are reporting the expression studies in yeast which has many added advantages like protein folding and processing. The expression studies were carried out in a new protein expression system based on the yeast Pichia pastoris called PichiaPink™. ansB gene isolated from E. coli which is coded for asparaginase was cloned into pPink HC-α plasmid and transformed into protease knock out pichia pink strain by electroporation. The recombinant enzyme was extracellular and showing the activity of 2.5 IU/ml. It was then purified using Ni-NTA column since the enzyme contains His-tag at the C-terminal end. The new way of expression would be efficient in making the enzyme humanized by glycosylation patterns which is similar to mammals.

Keywords
Enzyme technology; Auxotroph; Submerged culture; Recombinant DNA; Asparaginase; Pichia pink
First Page Preview
Cloning and expression of l-asparaginase from E. coli in eukaryotic expression system
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: Biochemical Engineering Journal - Volume 102, 15 October 2015, Pages 14–17
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