Summary
Many of the potential technical applications of alcohol oxidase (MOX; EC 1.1.3.13) are limited by the presence of high activities of catalase in the enzyme preparations. In order to circumvent laborious and costly purification or inactivation procedures, the induction of MOX in a catalase-negative mutant of Hansenula polymorpha has been studied. Emphasis was laid on the induction of activities of MOX and the dissimilatory enzymes in continuous cultures grown on various mixtures of formate/glucose and formaldehyde/glucose. In continuous cultures of the catalase-negative mutant grown on these mixtures, MOX can be induced efficiently. To obtain a stable and productive process, the ratio of the substrates is of critical importance. The optimal ratios of the mixtures for the catalase-negative strain for formate/glucose and formaldehyde/glucose were 3:1 and 1–2:1, respectively. Under identical cultivation conditions the wild-type strain showed similar induction patterns for MOX and the dissimilatory enzymes formaldehyde dehydrogenase (FaDH) and formate dehydrogenase (FoDH). The MOX levels in the catalase-negative strain were approx. 50% of those in the wild-type strain.
Similar content being viewed by others
References
Dijken van JP (1976) Thesis, University of Groningen
Dijken van JP, Otto R, Harder W (1976) Growth of Hansenula polymorpha in a methanol-limited chemostat. Physiological responses due to the involvement of methanol oxidase as a key enzyme in methanol metabolism. Arch Microbiol 111:137–144
Eggeling L, Sahm H, Wagner F (1977) Induction of FMN adenyltransferase in the methanol-utilizing yeast Candida boidinii. FEMS Microbiol Lett 1:205–210
Eggeling L, Sahm H (1980) Regulation of alcohol oxidase. Synthesis in Hansenula polymorpha: oversynthesis during growth on mixed substrates and induction by methanol. Arch Microbiol 127:119–124
Egli Th (1980) Wachstum von Methanol assimilierenden Hefen. Thesis ETH no: 6538, Zürich
Egli Th, Bosshard C, Hamer G (1986) Simultaneous utilization of methanol-glucose mixtures by Hansenula polymorpha in chemostat; influence of dilution rate and mixture composition on utilization pattern. Biotechnol Bioeng 28:1735–1741
Giuseppin MLF, Van Eijk HMJ, Bes BCM (1988) Molecular regulation of alcohol oxidase in Hansenula polymorpha in continuous cultures. Biotechnol Bioeng. In press
Lang E, Lang H (1972) Spezifische Farbereaktion zum direkten Nachweis der Ameisensäure. Z Anal Chem 260:8–10
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the Folin phenol reagent. J Biol Chem 193:265–271
Müller RH, Sysoev OV, Babel W (1986) Use of formate gradients for improving biomass yield of Pichia pinus growing continously on methanol. Appl Microbiol Biotechnol 25:238–244
Nash T (1953) The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J 55:416–421
Patel RN, Hou CT, Laskin AI, Deleranko P (1981) Microbial oxidation of methanol: properties of crystallized alcohol oxidase from a yeast Pichia sp. Arch Biochem Biophys 210:481–488
Shimuzu S, Ishida M, Tani Y, Ogata K (1977a) Derepression of FAD pyrophosphorylase and flavin changes during growth of Kloeckera sp. no. 2201 on methanol. Agric Biol Chem 11:2215–2220
Shimuzu S, Ishida M, Tani Y, Ogata K (1977b) Flavin changes of Kloeckera sp. no. 2201 during adaptation to methanol. Agric Biol Chem 41:423–424
Tani Y, Sakai Y, Yomada H (1985) Isolation and characterization of a mutant of a methanol yeast, Candida boidinii S2, with higher formaldehyde productivity. Agric Biol Chem 49:2699–2706
Unilever patent (1984) EPA 0173378
Unilever patent (1986) NL-8601454
Veenhuis M, Dijken van JP, Harder W (1983) The significance of peroxisomes in the metabolism of one-carbon compounds in yeast. Adv Microbiol Physiol 24:1–82
Verduyn C, Dijken van JP, Scheffers WA (1984) Colorimetric alcohol assays with alcohol oxidase. Int Microbiol Meth 42:15–25
Zwart KB, Harder W (1983) Regulation of the metabolism of some alkylated amines in the yeasts Candida utilis and Hansenula polymorpha. J Gen Microbiol 129:3159–3169
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Giuseppin, M.L.F., van Eijk, H.M.J., Verduyn, C. et al. Production of catalase-free alcohol oxidase by Hansenula polymorpha . Appl Microbiol Biotechnol 28, 14–19 (1988). https://doi.org/10.1007/BF00250490
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00250490