Yield, solubility and conformational quality of soluble proteins are not simultaneously favored in recombinant Escherichia coli.

Abstract:

:Many enzymes or fluorescent proteins produced in Escherichia coli are enzymatically active or fluorescent respectively when deposited as inclusion bodies. The occurrence of insoluble but functional protein species with native-like secondary structure indicates that solubility and conformational quality of recombinant proteins are not coincident parameters, and suggests that both properties can be engineered independently. We have here proven this principle by producing elevated yields of a highly fluorescent but insoluble green fluorescent protein (GFP) in a DnaK- background, and further enhancing its solubility through adjusting the growth temperature and GFP gene expression rate. The success of such a two-step approach confirms the independent control of solubility and conformational quality, advocates for new routes towards high quality protein production and intriguingly, proves that high protein yields dramatically compromise the conformational quality of soluble versions.

journal_name

Biotechnol Bioeng

authors

Martínez-Alonso M,García-Fruitós E,Villaverde A

doi

10.1002/bit.21996

subject

Has Abstract

pub_date

2008-12-15 00:00:00

pages

1353-8

issue

6

eissn

0006-3592

issn

1097-0290

journal_volume

101

pub_type

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