Cell wall strength of Hansenula polymorpha in continuous cultures in relation to the recovery of methanol oxidase (MOX)

The regulation of methanol oxidase (MOX) in Hansenula polymorpha has been studied in continuous cultures using a mixture of glucose and methanol (4:1 w/w) as carbon source. The study focused on the identification of stages in the biosynthesis affecting the formation of active MOX in glucose-methanol-grown cells. The levels of MOX mRNA, MOX protein in monomeric and octameric from, the ratio FAD/MOX, and the actual MOX activity have been quantified as functions of the dilution rate D. Hybridization studies with MOX mRNA probes showed an induction of MOX mRNA formation up to D = 0.29 h(-1). The induction of MOX protein synthesis (up to 37% of the cellular protein) is determined at low D values on the transcriptional level. The MOX activity at high D values is tuned by FAD incorporation and (post-) translation. Despite the high levels of MOX mRNA, decreasing levels of MOX activity and MOX protein were found at D values ranging from 0.14 t 0.29 h(-1). The maximal ratio FAD/MOX(6) was determined at D = 0.1 h(-1), which correlated with the maximal specific activity of MOX. In glucose-methanol media both protein level and MOX activity are repressed by increasing levels of residual glucose at high D values.

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Molecular regulation of methanol oxidase activity in continuous cultures of Hansenula polymorpha

Giuseppin

Eijk

Bes

1988

Biotech & Bioengineering

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Mathematical modeling of growth and production of alcohol oxidase by Hansenula polymorpha grown on methanol–glucose mixtures

Giuseppin

1989

Biotech & Bioengineering

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The modeling of growth and production of methanol oxidase (MOX) by Hansenula polymorpha CBS 4732 has been studied to provide a mathematical description of such production processes. Two kinds of mathematical models were constructed for growth on methanol and on mixtures of methanol and glucose. The model for growth on methanol as the sole carbon source consists of kinetics expressions, a limited number of key steps incorporating substrate and production inhibition. This model was used to predict and simulate the culture dynamics at the start-up, the most critical step in continuous cultivation. The growth on mixtures of methanol and glucose was modeled assuming virtually independent metabolic pathways. The induction and production of MOX could be described by adaptation of various repression equations for various data from the literature. The models describe both experimental data and literature data on growth of H. polymorpha CBS 4732 on glucose-methanol mixtures satisfactorily. All parameters for the induction-repression model for growth of H. polymorpha CBS 4732 on glucose-methanol mixtures yielded evidence that a similar induction-repression pattern is involved in MOX production. Catalase, however, is repressed by a different mechanism.

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Rapid transformation of nor-Saccharomyces yeasts by electroporation

1993

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Cell wall strength of Hansenula polymorpha in continuous cultures in relation to the recovery of methanol oxidase (MOX)

Cited by 8 publications

References 9 publications

Molecular regulation of methanol oxidase activity in continuous cultures of Hansenula polymorpha

Molecular regulation of methanol oxidase activity in continuous cultures of Hansenula polymorpha

Mathematical modeling of growth and production of alcohol oxidase by Hansenula polymorpha grown on methanol–glucose mixtures

Rapid transformation of nor-Saccharomyces yeasts by electroporation

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