Masayuki Onodera

Sterol Biosynthesis by a Prokaryote: First in Vitro Identification of the Genes Encoding Squalene Epoxidase and Lanosterol Synthase from Methylococcus capsulatus

Sterol biosynthesis by prokaryotic organisms is very rare. Squalene epoxidase and lanosterol synthase are prerequisite to cyclic sterol biosynthesis. These two enzymes, from the methanotrophic bacterium Methylococcus capsulatus, were functionally expressed in Escherichia coli. Structural analyses of the enzymatic products indicated that the reactions proceeded in a complete regio- and stereospecific fashion to afford (3S)-2,3-oxidosqualene from squalene and lanosterol from (3S)-2,3-oxidosqualene, in full accordance with those of eukaryotes. However, our result obtained with the putative lanosterol synthase was inconsistent with a previous report that the prokaryote accepts both (3R)- and (3S)-2,3-oxidosqualenes to afford 3-epi-lanosterol and lanosterol, respectively. This is the first report demonstrating the existence of the genes encoding squalene epoxidase and lanosterol synthase in prokaryotes by establishing the enzyme activities. The evolutionary aspect of prokaryotic squalene epoxidase and lanosterol synthase is discussed.

Silicone rubber membrane bioreactors for bacterial cellulose production

Cellulose production byAcetobacter pasteurianus was investigated in static culture using four bioreactors with silicone rubber membrane submerged in the medium. The shape of the membrane was flat sheet, flat sack, tube and cylindrical balloon. Production rate of cellulose as well as its yield on consumed glucose by the bacteria grown on the flat type membranes was approximately ten-fold greater than those on the non-flat ones in spite of the same membrane thickness. The membrane reactor using flat sacks of silicone rubber membrane as support of bacterial pellicle can supply greater ratio of surface to volume than a conventional liquid surface culture and is promising for industrial production of bacterial cellulose in large scale.

Accumulation of S-2-hydroxyethyl derivatives of L-cysteine and L-homocysteine by a methionine auxotroph of Escherichia coli with the addition of 2-mercaptoethanol to the culture.

We previously reported [J. Biosci. Bioeng., 94, 178-181 (2002)] that an Escherichia coli MetC-deficient mutant can accumulate L-cystathionine. When 2-mercaptoethanol was added to the culture medium during fermentation, the accumulation of L-cystathionine decreased and S-(2-hydroxyethyl)-L-cysteine and S-(2-hydroxyethyl)-L-homocysteine were accumulated.