Pyrroloquinoline quinone biosynthesis in Escherichia coli through expression of the Gluconobacter oxydans pqqABCDE gene cluster
Tóm tắt
We have expressed the pqqABCDE gene cluster from Gluconobacter oxydans, which is involved in pyrroloquinoline quinone (PQQ) biosynthesis, in Escherichia coli, resulting in PQQ accumulation in the medium. Since the gene cluster does not include the tldD gene needed for PQQ production, this result suggests that the E. coli tldD gene, which shows high homology to the G. oxydans
tldD gene, carries out that function. The synthesis of PQQ activated d-glucose dehydrogenase in E. coli and the growth of the recombinant was improved. In an attempt to increase the production of PQQ, which acts as a vitamin or growth factor, we transformed E. coli with various recombinant plasmids, resulting in the overproduction of the PQQ synthesis enzymes and, consequently, PQQ accumulation—up to 6 mM—in the medium. This yield is 21.5-fold higher than that obtained in previous studies.
Tài liệu tham khảo
Duine JA, Jongejan JA (1989) Quinoproteins, enzyme with pyrrolo-quinoline quinone as cofactor. Annu Rev Biochem 58:403–426
Matsushita K, Toyama H, Yamada M, Adachi O (2002) Quinoproteins: structure, function, and biotechnological applications. Appl Microbiol Biotechnol 58:13–22
Goodwin PM, Anthony C (1998) The biochemistry, physiology and genetics of PQQ and PQQ-containing enzymes. Adv Microb Physiol 40:1–80
Matsushita K, Arents JC, Bader R, Yamada M, Adachi O, Postma PW (1997) Escherichia coli is unable to produce pyrroloquinoline quinone (PQQ). Microbiology 143:3149–3156
Meulenberg JJ, Sellink E, Riegman NH, Postma PW (1992) Nucleotide sequence and structure of the Klebsiella pneumoniae pqq operon. Mol Gen Genet 232:284–294
Morris CJ, Biville F, Turlin E, Lee E, Ellermann K, Fan WH, Ramamoorthi R, Springer AL, Lidstrom ME (1994) Isolation, phenotypic characterization, and complementation analysis of mutants of Methylobacterium extorquens AM1 unable to synthesize pyrroloquinoline quinone and sequences of pqqD, pqqG, and pqqC. J Bacteriol 176:1746–1755
Springer AL, Ramamoorthi R, Lidstrom ME (1996) Characterization and nucleotide sequence of pqqE and pqqF in Methylobacterium extorquens AM1. J Bacteriol 178:2154–2157
Zhang M, Lidstrom ME (2003) Promoters and transcripts for genes involved in methanol oxidation in Methylobacterium extorquens AM1. Microbiology 149:1033–1040
Gliese N, Khodaverdi V, Schobert M, Görisch H (2004) AgmR controls transcription of a regulon with several operons essential for ethanol oxidation in Pseudomonas aeruginosa ATCC 17933. Microbiology 150:1851–1857
Felder M, Gupta A, Verma V, Kumar A, Qazi GN, Cullum J (2000) The pyrroloquinoline quinone synthesis genes of Gluconobacter oxydans. FEMS Microbiol Lett 193:231–236
Prust C, Hoffmeister M, Liesegang H, Wiezer A, Fricke WF, Ehrenreih A, Gottschalk G, Deppenmeier U (2005) Complete genome sequence of the acetic bacterium Gluconobacter oxydans. Nat Biotechnol 23:195–200
Hölscher T, Görisch H (2006) Knockout and overexpression of pyrroloquinoline quinone biosynthetic genes in Gluconobacter oxydans 621H. J Bacteriol 188:7668–7676
Goosen N, Horsman HP, Huinen RG, Van de Putte P (1989) Acinetobacter calcoaceticus genes involved in biosynthesis of the coenzyme pyrrolo-quinoline-quinone: nucleotide sequence and expression in Escherichia coli K-12. J Bacteriol 171:447–455
Meulenberg JJM, Sellink E, Loenen WAM, Riegman NH, Van Kleef M, Postma PW (1990) Cloning of Klebsiella pneumoniae. pqq genes and PQQ biosynthesis in Escherichia coli. FEMS Microbiol Lett 71:337–344
Yang XP, Wei LJ, Lin JP, Wei DZ (2008) Membrane-bound pyrroloquinoline quinone-dependent dehydrogenase in Gluconobacter oxydans M5, responsible for production of 6-(2-hydroxyethyl) amino-6-deoxy-l-sorbose. Appl Environ Microbiol 74:5250–5253
Bradford MM (1976) A rapid and simple method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Geiger O, Görisch H (1987) Enzymatic determination of pyrroloquinoline quinone using crude membranes from Escherichia coli. Anal Biochem 164:418–423
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor
Adamowicz M, Conway T, Nickerson KW (1991) Nutritional complementation of oxidative glucose metabolism in Escherichia coli via pyrroloquinoline quinone-dependent glucose dehydrogenase and the Entner–Doudoroff pathway. Appl Environ Microbiol 57:2012–2015
Fliege R, Tong S, Shibata A, Nickerson KW, Conway T (1992) The Entner-Doudoroff pathway in Escherichia coli is induced for oxidative glucose metabolism via pyrroloquinoline quinone-dependent glucose dehydrogenase. Appl Environ Microbiol 58:3826–3829
Steinberg F, Stites TE, Anderson P, Storms D, Chan I, Eghbali S, Rucker RB (2003) Pyrroloquinoline quinone improves growth and reproductive performance in mice fed chemically defined diets. Exp Biol Med 228:160–166