Bioconversion of Methanol to Formaldehyde: 1 by Free Whole Cell and Immobilized Whole Cell of YeastHansenula polymorpha

Abstract

Modified methylotrophic yeast Hansenula polymorpha (HP A16) was used in this study, which was obtained with recrossing a leucine oxotrophic yeast and a wild type Hansenula polymorpha CB4732 and was discovered by Ogata et al. The yeast is grown with methanol as a sole carbon source in which methanol oxidase (MOX) is a key enzyme of methanol metabolism. Because of its stability and low substrate specificity, alcohol oxidase is of considerable interest for a range of biotechnological processes. Various methanol feeding regimes were evaluated in an effort to increase the biomass concentration and productivity that could be achieved from fermentations using the other Hansenula polymorpha species. This yeast was grown by batch fermentation. The effects of conditions of inoculation media for increasing amount of MOX enzyme in peroksizomes of yeast with MOX activity were observed. The highest MOX activity of yeast was found within optic density of grown media of OD600 1.5, at 0.35 microM of methanol as used an oxotrophic substrate, at 35 degrees C temperature, at pH 7.0 of 0.1 M potassium phosphate buffer (KPB), at 40 microL of buffered yeast cell volume and at the incubation time of 50 minutes. Whole yeast cells were cultivated at above optimized incubation conditions. The cells were immobilized within the polyacyrlamide gels by entrapment method. Free whole cells and immobilized whole cells were compared using bioconversion percentages of methanol to formaldehyde.