Summary
A selection by glucosamine for mutants of Hansenula polymorpha insensitive to glucose repression of methanol assimilation is described. Constitutive synthesis of enzymes is established in standard batch cultures of glucosegrown cells. Upon prolonged glucose metabolism the phenotype is masked by catabolite inactivation and degradation of enzymes. Addition of the substrate methanol remarkably improves constitutive synthesis by preventing catabolite inactivation and delaying degradation. Regular peroxisomes of reduced number are formed in mutant cells under repressed conditions. No constitutive synthesis is detectable using ethanol as a carbon source. In addition, this alcohol is detrimental to growth of the mutants, indicating that H. polymorpha is constrained to repress synthesis of enzymes involved in the C1-metabolism when ethanol is present as a substrate.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bormann C, Sahm H (1978) Degradation of microbodies in relation to activities of alcohol oxidase and catalase in Candida boidinii. Arch Microbiol 117:67–72
Bruinenberg PG, Veenhuis M, van Dijken JP, Duine JA, Harder W (1982) A quantitative analysis of selective inactivation of peroxisomal enzymes in the yeast Hansenula polymorpha by high-performance liquid chromatography. FEMS Microbiol Lett 15:45–50
Celenza JL, Carlson M (1986) A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science 233:1175–1180
Chrastil J, Wilson JT (1975) A sensitive colorimetric method for formaldehyde. Anal Biochem 63:202–207
de Duve C, Baudhuin P (1966) Peroxisomes (microbodies and related particles). Physiol Rev 46:323–357
Eggeling L, Sahm H (1978) Derepression and partial insensitivity to carbon catabolite repression of the methanol-dissimilating enzymes in Hansenula polymorpha. Eur J Microbiol Biotechnol 5:197–202
Entian KD (1987) Glucose repression: a complex regulatory system in yeast. Microb Sci 3:366–371
Fukui S, Kawamoto S, Yasuhara S, Tanaka A, Osumi M, Imaizumi F (1975) Microbody of methanol-grown yeasts. Localization of catalase and flavin-dependent alcohol oxidase in the isolated microbody. Eur J Biochem 59:561–566
Holzer H (1976) Catabolite inactivation in yeast. Trends Biochem Sci 1:178–181
Lazarow PB, Fujiki Y (1985) Biogenesis of peroxisomes. Annu Rev Cell Biol 1:489–530
Roggenkamp R, Sahm H, Wagner F (1974) Microbial assimilation of methanol. Induction and function of catalase in Candida boidinii. FEBS Lett 41:283–286
Roggenkamp R, Sahm H, Hinkelmann W, Wagner F (1975) Alcohol oxidase and catalase in peroxisomes of methanol-grown Canadida boidinii. Eur J Biochem 59:637–646
Roggenkamp R, Janowicz Z, Stanikowski B, Hollenberg CP (1984) Biosynthesis and regulation of the peroxisomal methanol oxidase from the methylotrophic yeast Hansenula polymorpha. Mol Gen Genet 194:489–493
Roggenkamp R, Hansen H, Eckart M, Janowicz Z, Hollenberg CP (1986) Transformation of the methylotrophic yeast Hansenula polymorpha by autonomous replication and integration vectors. Mol Gen Genet 202:302–308
Sahm H, Wagner F (1973) Microbial assimilation of methanol. The ethanol-and methanol oxidizing enzymes of the yeast Candida boidinii. Eur J Biochem 36:250–256
Sahm H, Roggenkamp R, Hinkelmann W, Wagner F (1975) Microbodies in methanol-grown Candida boidinii. J Gen Microbiol 88:218–222
Sakai Y, Sawai T, Tani Y (1987) Isolation and characterization of a catabolite repression-insensitive mutant of a methanol yeast, Candida boidinii A5, producing alcohol oxidase in glucose-containing medium. Appl Environ Microbiol 53:1812–1818
Schütte H, Flossdorf J, Sahm H, Kula MR (1976) Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii. Eur J Biochem 62:151–160
Tanaka O, Osumi M, Fukui S (1982) Peroxisomes of alkane-grown yeast: Fundamental and practical aspects. Ann NY Acad Sci 386:183–192
Tolbert NE (1981) Metabolic pathways in peroxisomes and glyoxysomes. Annu Rev Biochem 50:133–157
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some application. Proc Natl Acad Sci USA 76:4350–4354
Veenhuis M, Harder W (1987) Metabolic significance and biogenesis of microbodies in yeast. In: Fahimi HD, Sies H (eds) Peroxisomes in biology and medicine. Springer, Berlin Heidelberg New York, p 437
Veenhuis M, van Dijken JP, Harder W (1983) The significance of peroxisomes in the metabolism of one carbon compounds in yeasts. Adv Microb Physiol 24:1–82
Veenhuis M, Mateblowski M, Kunau WH, Harder W (1987) Proliferation of microbodies in Saccharomyces cerevisiae. Yeast 3:77–84
Zubay G, Schwartz D, Beckwith J (1970) Mechanism of activation of catabolite sensitive genes: a positive control system. Proc Natl Acad Sci USA 66:104–110
Author information
Authors and Affiliations
Additional information
Communicated by R.G. Herrmann
Rights and permissions
About this article
Cite this article
Roggenkamp, R. Constitutive appearance of peroxisomes in a regulatory mutant of the methylotrophic yeast Hansenula polymorpha . Mol Gen Genet 213, 535–540 (1988). https://doi.org/10.1007/BF00339627
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00339627