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Regulatory flexibility of methylotrophic yeasts in chemostat cultures: Simultaneous assimilation of glucose and methanol at a fixed dilution rate

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Abstract

The influence of the composition of methanol/glucose-mixtures as only sources of carbon and energy on growth and regulation of the synthesis of enzymes involved in methanol-dissimilation was studied under chemostat conditions at a fixed dilution rate with the methylotrophic yeasts Hansenula polymorpha and Kloeckera sp. 2201. Both carbon sources were found to be utilized completely independently of the composition of the C1/C6 mixture. Using mixtures of 14C-labelled methanol and glucose the growth yield for glucose was found to be constant for all C1/C6-mixtures tested and both yeasts. The growth yield for methanol, however, was reduced by up to 25% when the proportion of methanol in the inflowing medium was lower than 20% (w/w with respect to glucose) for H. polymorpha and 50% (w/w with respect to glucose) for Kloeckera sp. 2201 respectively. During growth with C1/C6-mixtures containing higher C1-proportions of methanol regular growth yields for methanol were recorded which corresponded to the growth yields found with methanol as the only carbon source.

The regulation of the synthesis of the enzymes of the dissimilatory pathway for methanol was found to be under multiple control. Although glucose was present in the medium methanol had a positive effect on the synthesis of these enzymes. Thus, in addition to derepression induction by methanol was also observed. This inductive effect was found to increase with increasing proportions of methanol in the mixture. Depending on the enzyme, 10–40% methanol in the mixture resulted in a maximal induction with enzyme specific activities equal to those found in cells grown with methanol as the only carbon source. No further enhancements in enzyme specific activities were observed during growth on mixtures containing more than 40% methanol.

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Abbreviations

C1 :

Methanol

C6 :

glucose

C1/C6 :

mixture compositions are given in % (w/w)

C0 :

concentration of 14C in the inflowing medium (DPM ml-1)

C(t):

concentration of 14C incorporated in cells as a function of time t (DPM ml-1)

d :

dilution rate (h-1)

DPM:

disintegrations per minute

q s :

q C1 and q C6 are specific rates of consumption of substrate, methanol and glucose respectively [g (g cell dry weight)-1 h-1]

q O2 and q CO2 :

are the specific rates of oxygen consumption and carbon dioxide release [mmol (g cell dry weight)-1 h-1]

RQ:

respiration quotient (q CO2 q O2 -1)

s C1 and s C6 :

are the residual concentrations of methanol and glucose in the culture liquid (g l-1)

s O/C1 and s O/C6 :

are the concentrations of methanol and glucose in the inflowing medium (g l-1)

Sp.A.:

enzyme specific activity

x :

cell dry weight concentration (g l-1)

Y X/C1 and Y X/C6 :

are growth yields on methanol and glucose respectively (g cell dry weight (g substrate)-1

Y C/C1 :

growth yield with methanol with respect to carbon (g carbon assimilated (g carbon supplied)-1

μm :

maximum specific growth rate (h-1)

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Egli, T., Käppeli, O. & Fiechter, A. Regulatory flexibility of methylotrophic yeasts in chemostat cultures: Simultaneous assimilation of glucose and methanol at a fixed dilution rate. Arch. Microbiol. 131, 1–7 (1982). https://doi.org/10.1007/BF00451490

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Key words

  • Hansenula polymorpha
  • Kloeckera sp. 2201
  • Chemostat
  • Mixed substrates
  • Glucose
  • Methanol
  • Growth yields
  • Enzyme regulation
  • Dissimilatory enzymes