Summary
In methanol-utilizing yeasts, catalase is an essential enzyme for the destruction of hydrogen peroxide generated by methanol oxidase (E.C. 1.1.3.13). It was found however that a catalase-negative mutant of Hansenula polymorpha is able to consume methanol in the presence of glucose in continuous cultures. At a dilution rate of 0.1 h-1, stable continuous cultures could be obtained during growth on methanol/glucose mixtures with a molar ratio of methanol/glucose between 0 to 2.4. In these cultures methanol oxidase was induced up to a level of 40% of that obtained in the wild-type strain. The hydrogen peroxide-decomposition activity of the mutant was studied in more detail by pulsing methanol to samples of steady-state cultures. Only after the addition of excess methanol the hydrogen peroxide-decomposing system became saturated, and the cells excreted hydrogen peroxide. This was accompanied by excretion of formaldehyde and a rapid loss of viability. The presence of extracellular catalase during a methanol pulse prevented the loss of viability. The nature of the alternative hydrogen peroxide-decomposing enzyme system remains to be elucidated. Its capacity strongly depended on the cultivation conditions and pretreatment of the cells. Cells grown on formaldehyde/glucose mixtures showed a lower methanol tolerance than those grown on the methanol/glucose mixtures. Freeze-drying of cells drastically enhanced the excretion of hydrogen peroxide, probably as a result of an inactivation of the decomposing system.
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Giuseppin, M.L.F., van Eijk, H.M.J., Bos, A. et al. Utilization of methanol by a catalase-negative mutant of Hansenula polymorpha . Appl Microbiol Biotechnol 28, 286–292 (1988). https://doi.org/10.1007/BF00250457
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DOI: https://doi.org/10.1007/BF00250457