Low-solubility glycerol dehydratase, a chimeric enzyme of coenzyme B12 -dependent glycerol and diol dehydratases.
Coenzyme B(12)-dependent diol and glycerol dehydratases are isofunctional enzymes, which catalyze dehydration of 1, 2-diols to produce corresponding aldehydes. Although the two types of dehydratases have high sequence homology, glycerol dehydratase is a soluble cytosolic enzyme, whereas diol dehydratase is a low-solubility enzyme associated with carboxysome-like polyhedral organelles. Since both the N-terminal 20 and 16 amino acid residues of the beta and gamma subunits, respectively, are indispensable for the low solubility of diol dehydratase, we constructed glycerol dehydratase-based chimeric enzymes which carried N-terminal portions of the beta and gamma subunits of diol dehydratase in the corresponding subunits of glycerol dehydratase. Addition of the diol dehydratase-specific N-terminal 34 and 33 amino acid residues of the beta and gamma subunits, respectively, was not enough to lower the solubility of glycerol dehydratase. A chimeric enzyme which carries the low homology region (residues 35-60) of the diol dehydratase beta subunit in addition to the diol dehydratase-specific extra-regions of beta and gamma subunits showed low solubility comparable to diol dehydratase, although its hydropathy plot does not show any prominent hydrophobic peaks in these regions. It was thus concluded that short N-terminal sequences are sufficient to change the solubility of the enzyme.
Tobimatsu
Takamasa
T
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama 700-8530, Japan.
Nishiki
Tsuneo
T
Morimoto
Masaya
M
Miyata
Ryou
R
Toraya
Tetsuo
T
D000595:Amino Acid Sequence; D003038:Cobamides / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D006836:Hydro-Lyases / Q000737:chemistry*; D008969:Molecular Sequence Data; D011408:Propanediol Dehydratase / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility
D003038:Cobamides; D011994:Recombinant Proteins; D006836:Hydro-Lyases; D011408:Propanediol Dehydratase; C020671:glycerol dehydratase; C000913:cobamamide