Synthesis and assembly of a novel recombinant ribulose bisphosphate carboxylase/oxygenase

Journal Article · Sat Feb 01 04:00:00 EST 1986 · Bio/Technology; (United States)

DOI:https://doi.org/10.1038/nbt0286-138· OSTI ID:6000262

Tabita, F R; Gibson, J L; Mandy, W J; Quivey, Jr, R G

The Rhodopseudomonas sphaeroides form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPC/O) expressed in Escherichia coli has been purified to electrophoretic homogeneity by dye-ligand column chromatography. The form II RuBPC/O made in E. coli exactly co-migrates with the R. sphaeroides enzyme in sodium dodecyl sulfate polyacrylamide slab gels. Immunological identity of the recombinant protein was verified by Western immunoblot analysis using antiserum directed against form II RuBPC/O. In addition, sequence determination of amino terminus of the cloned gene product has established that the protein produced in E. coli under lac transcriptional control is identical to the R. sphaeroides enzyme. Based on comparisons of various catalytic properties, the RuBPC/O isolated from E. coli is functionally identical to the R. sphaeroides enzyme. The E. coli form II RuBPC/O should thus serve as an excellent model for subsequent molecular manipulation of this agriculturally important protein.

• Research Organization: Univ. of Texas, Austin
• OSTI ID: 6000262
• Journal Information: Bio/Technology; (United States), Journal Name: Bio/Technology; (United States) Vol. 4:2; ISSN BTCHD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
BACTERIA
BIOLOGICAL MODELS
CHROMATOGRAPHY
ELECTROPHORESIS
ENZYMES
ESCHERICHIA COLI
GENES
MICROORGANISMS
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
OXIDOREDUCTASES
OXYGENASES
PROTEINS
PURIFICATION
RHODOPSEUDOMONAS
SEPARATION PROCESSES