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  • 1
    ISSN: 1432-072X
    Keywords: Oxidative phosphorylation ; Chemostat culture ; Growth yield ; Mixed substrates ; Ribulose-bisphosphate cycle ; Cytochrome c ; Single cell protein ; Methanol ; Paracoccus denitrificans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Paracoccus denitrificans was grown aerobically during two-(carbon)substrate-limitation on mannitol and methanol in chemostat cultures. Theoretical growth parameters were calculated based on the presence of 2 or 3 sites in the electron-transport chain of Paracoccus denitrificans. Experimental growth parameters determined during two-(carbon)substrate growth were conform to the presence of 3 sites of oxidative phosphorylation, while cells grown only on mannitol possessed 2 sites. The maximum growth yield on adenosine triphosphate (ATP), corrected for maintenance requirements, determined in chemostat experiments in which the methanol concentration is less than 2.11 times the mannitol concentration was 8.6 g of biomass. When the methanol concentration was more than 2.11 times the mannitol concentration the maximum growth yield on adenosine triphosphate decreased due to the more energy consuming process of CO2-assimilation. Cells use methanol only as energy source to increase the amount of mannitol used for assimilation purposes. When the methanol concentration in chemostat experiments was more than 2.11 times the mannitol concentration, all mannitol was used for assimilation and excess energy derived from methanol was used for CO2-assimilation via the ribulose-bisphosphate cycle. The synthesis of ribulosebisphosphate carboxylase was repressed when the methanol concentration in chemostat experiments was less than 2.11 times the mannitol concentration or when Paracoccus denitrificans was grown in batch culture on both methanol and mannitol. When in chemostat experiments the methanol concentration was more than 2.11 times the mannitol concentration ribulose-bisphosphate carboxylase activity could be demonstrated and CO2-assimilation will occur. It is proposed that energy produced in excess activates or derepresses the synthesis of the necessary enzymes of the ribulose-bisphosphate cycle in Paracoccus denitrificans. Consequently growth on any substrate will be carbonas well as energy-limited. When methanol is present in the nutrient cells of Paracoccus denitrificans synthesize a CO-binding type of cytochrome c, which is essential for methanol oxidase activity. The reason for the increase in efficiency of oxidative phosphorylation from 2 to 3 sites is most probably the occurrence of this CO-binding type of cytochrome c in which presence electrons preferentially pass through the a-type cytochrome region of the electron-transport chain.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1572-9699
    Keywords: Aspergillus ; continuous culture ; fungi ; protein production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspergillus niger has been grown in glucose- and maltose-limited continuous cultures to determine the bioenergetic consequences of the production of the extracellular enzyme glucoamylase. Growth yields (g biomass per mol substrate) were high, indicating that growth was very efficient and protein production for biomass was not exceedingly energy consuming. It has been found that the energy costs for the production of this extracellular enzyme is very high. Depending on the efficiency of energy conservation the glucoamylase protein yield on ATP is between 1.3 and 2.6 g protein per mol ATP, which is equal or less than 10% of the theoretical maximum of 25.5. These high energy costs most probably have to be invested in the process of excretion. A comparison between an industrial over-producing strain and the wild typeAspergillus niger showed that this over-producing strain most probably is a regulatory mutant. Two regions of specific growth rates could be determined (one at specific growth rates lower and one at specific growth rates higher than 0.1 h-1), which are characterized by differences in mycelium morphology and a significant deviation from linearity in the linear equation for substrate utilization. Analysis of the region of specific growth rates higher than 0.1 h-1 yielded maintenance requirements of virtual zero. It has been concluded that for a good analysis of the growth behaviour of filamentour fungi the linear equation for substrate utilization is not suitable, since it contains no term for the process of differentiation.
    Type of Medium: Electronic Resource
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