Alcohol Pichia pastoris

Dienys, G., Jarmalavicius, S., Budriene, S. et al. (2003) Alcohol oxidase from the yeast Pichia pastoris — a potential catalyst for organic synthesis. Journal of Molecular Catalysis B-Enzymatic, 21 (1-2), 47—49. 

Kawakami, K. and Furukawa, S.Y. (1997) Alcohol-oxidation activity of whole cells of Pichia pastoris entrapped in hybrid gels composed of Ca-alginate and organic silicate. Applied Microbiology and Biotechnology, 67, 23-31. 

Scheme 23.2 Production of aliphatic flavour aldehydes from natural alcohols using alcohol oxidase activity of Pichia pastoris cells...

Benzyldehyde Benzoic acid, benzyl alcohol or phenylalanine Polyporus tuberaster, Pichia pastoris Almond... 

By utilizing an alcohol oxidase from Pichia pastoris or Candida sp.11481, almost complete conversion of ethylene glycol into glyoxal (Fig. 16.2-30) was observed. These enzymatic routes were shown to be superior in terms of reaction conditions and yields compared to the chemical variants that make use of metal catalysts or even nitric acid for the oxidation of ethylene glycol. 

Cos O, Resina D, Ferrer P et al. (2005) Heterologous production of Rhizopus oryzae lipase in Pichia pastoris using the alcohol oxidase and formaldehyde dehydrogenase promoters in batch and fed-batch cultures. Biochem Eng J 26 86-94 Cowan D (1996) Industrial enzyme technology. TIBTECH 14 177-178... 

Functional characterization of the two alcohol oxidase genes from the yeast Pichia pastoris. Mol. Cell Biol, 9, 1316-1323. 

Alcohols Lower alcohols have been measured with alcohol oxidase (EC 1.1.3.13) from Candida boidinii or Pichia pastoris. The latter is available with a higher specific activity and has a somewhat different substrate specificity. Coimmobilization with catalase increases the stability of the enzyme column to several months with an operating range of 0.005-1 mmol 1 (0.5 ml samples) using 0.1 mol 1 sodium phosphate, pH 7.0, as the buffer. This assay is useful for the determination of ethanol in samples from beverages, blood, and for monitoring fermentation. 

Duff SJB, Murray WD (1990) Non aqueous reaction systems for the oxidation of higher molecular weight alcohols by alcohol oxidase from Pichia pastoris. In Charalambous G (ed) Flavours and off-flavours 89. Developments in food science, vol 24. Elsevier, Amsterdam, pp 701-713 Esguerra EB, Kawada K, Kitagawa H (1992) Removal of astringency in Amas Banana (Musa AA group) with postharvest ethanol treatment. Acta Hortic 321 811-820 Eskin NAM (1979) Terpenoides and flavonoides. In Plant pigments, flavours and textures. Academic Press, New York, pp 65-93... 

A very similar process is used to catalyze the oxidation of aromatic and aliphatic alcohols to their corresponding aldehydes. An alcohol oxidase enzyme isolated from Pichia pastoris placed in a two-phase system will readily produce benzaldehyde from benzyl alcohol. While this oxidation is slow and inefficient in aqueous systems, oxidation in a two-phase system increased yield by a factor of nine and reaction time was shortened greatly [74]. 

Duff, S.J.B., W.D. Murray, Oxidation of benzyl alcohol by whole cells of Pichia pastoris and by alcohol oxidase in aqueous and nonaqueous reaction media, Biotech-nol. Bioeng., 34, p. 153, 1989. 

Benzyl alcohol has also been employed as substrate with the methylotrophic yeast Pichia pastoris, in the presence of methanol, that induces the production of nonspecific alcohol oxidases able to convert benzyl alcohol to benzaldehyde. Very recently, in situ product removal approaches (adsorption and/or pervaporation) have been exploited with this system. In particular, adsorption on hydrophobic resin in a solid-liquid two-phase partitioning bioreactor afforded yields of up to 14g in a 5-1 reactor [104,105]. 

In order to avoid the use of alanine, an expensive reactant, thioanisole, was selectively oxidized to (S)-methyl phenyl sulfoxide in a 1.0 g scale with a 72% yield and 75% ee after 24 h by using the alcohol oxidase from Pichia pastoris. This flavoprotein oxidase was able to oxidize methanol to formaldehyde, generating hydrogen peroxide, which is employed by CiP [20]. 

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