Ribulose-bisphosphate carboxylase manipulation in the hydrogen bacterium Alcaligenes eutrophus

Abstract

29 separated by an intervening stretch of DNA, variable in length depending on the particular species of origin (Curtis & Hazelkorn, 1983; Shinozaki & Sugiura, 1983). The gene from S,.rzc~c.lioc.oc~r~us PCC30 1 (Reichelt & Delaney, 1983) has been cloned into the ampr region of PLa2311 (Remaut ct al., 1981) with expression controlled by the strong temperature-sensitive leftward promoter (PI) of bacteriophage EL (Gatenby et al., 1985). The construct is expressed in E. coli host organisms and produces active Rubisco. At present the levels of enzyme are poor, with low specific activity, possibly due to an under-production of the S-subunit. However, the synthesis of an active, multisubunit enzyme in non-photosynthetic bacteria argues against the necessity of a complicated assembly mechanism for Rubisco from cyanobacteria. Better constructs are certain to be developed that can be manipulated by 'in vitro' techniques and produce large amounts of enzymes. These will then be exploited to determine the structural basis for the differences between Rubisco from diverse photosynthetic organisms and indicate ways of changing the partition coefficient of the bisphosphate substrate.

Molecular and cellular regulation of autotrophic carbon dioxide fixation in microorganisms.

“…Alcaligenes eutrophus H16 contains a self-transmissible 450-kb megaplasmid (pHG1) which contains both the soluble and membrane-bound hydrogenases required for autotrophic growth (88, 90). Further analysis indicated that both pHG1 and the chromosome encode copies of the rbcL rbcS and prk genes in precisely the same organization (6,37,166). However, unlike the situation in Rhodobacter sphaeroides, both the plasmid and chromosomal prk genes are found downstream from the rbcL rbcS sequences at precisely the same location (37).…”

Sequence analysis of the Alcaligenes eutrophus chromosomally encoded ribulose bisphosphate carboxylase large and small subunit genes and their gene products

“…Here we report our analysis of the complete sequence of the rbcL and rbcS genes. A preliminary account of some of the results presented here was given previously (5).…”

Metabolic and Molecular Regulation of the CO2-Assimilating Enzyme System in Aerobic Chemoautotrophs

“…eutrophus ATCC 17707 [48,49]. Further studies on this system will certainly contribute to the elucidation of the complex genetic regulation of autotrophic C0 2 fixation in a facultative autotroph.…”