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Overexpression of a heterologous protein, haloalkane dehalogenase, in a poly‐β‐hydroxybutyrate‐deficient strain of the facultative methylotroph Methylobacterium extorquens AM1
Author(s) -
Fitzgerald Kelly A.,
Lidstrom Mary E.
Publication year - 2002
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10470
Subject(s) - heterologous , heterologous expression , biochemistry , strain (injury) , green fluorescent protein , chemistry , expression vector , lac operon , mutant , wild type , biology , recombinant dna , microbiology and biotechnology , gene , anatomy
Using an expression vector containing p mxaF , a strong native promoter, expression of a model heterologous protein, haloalkane dehalogenase, from Xanthobacter autotrophicus GJ10 was achieved in the methylotrophic bacterium, Methylobacterium extorquens AM1. Although expression using the wild‐type strain was <5% of total cell protein, expression at a level of 10% of the total cell protein was achieved in a mutant unable to synthesize poly‐β‐hydroxybutyrate granules. Two other tested heterologous proteins, catechol dioxygenase and green fluorescent protein, were expressed at moderate levels in both wild‐type and the PHB‐negative strain. These results suggest that the M. extorquens PHB‐negative strain is a possible platform for overexpression of heterologous proteins with labeled or unlabeled methanol as a starting material. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 263–268, 2003.