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Expression and in situ processing of human prorenin to active renin in baculovirus-infected Sf-9 insect cell cultures under several infective conditions

Paper ID Volume ID Publish Year Pages File Format Full-Text
4303 219 2009 5 PDF Available
Title
Expression and in situ processing of human prorenin to active renin in baculovirus-infected Sf-9 insect cell cultures under several infective conditions
Abstract

In the baculovirus expression vector system (BEVS), intrinsic proteases concomitantly produced by infected insect cells have been generally regarded as a defect, because they sometimes degrade expressed recombinant proteins and decrease the productivity. The present study successfully used the proteolysis to generate active recombinant human- (rh) renin after the expression of inactive rh-prorenin. Sf-9 insect cells were infected with recombinant baculoviruses having human preprorenin cDNA in the site of polyhedron gene at several MOIs. At any MOIs, rh-prorenin was expressed in a late phase of infective cultures and processed to active rh-renin in a very late phase. The maximum volumetric yield of active rh-renin was obtained at MOIs of 1 and 10 pfu/cell. The protease activity was examined with an internally quenched fluorogenic substrate newly designed for the processing. The generation of rh-renin was coincided with a considerable increase in a protease activity that was classified into the cysteine protease family, and significantly suppressed by supplementing the culture medium with leupeptin, a cysteine protease inhibitor. This suggested that the cysteine protease was responsible to the processing of rh-prorenin to rh-renin.

Keywords
Sf-9 insect cell; Baculovirus; Protease; Renin; Prorenin
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Expression and in situ processing of human prorenin to active renin in baculovirus-infected Sf-9 insect cell cultures under several infective conditions
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 43, Issue 2, 15 February 2009, Pages 216–220
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