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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 136 (1983), S. 203-208 
    ISSN: 1432-072X
    Keywords: Maximum growth yield ; Mixed substrate utilization ; Glucose ; Formate ; Transient-state cultivation ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The experimentally determined growth yield on glucose under aerobic conditions is approximately 0.5 g/g, but on the basis of the carbon content a value of 0.71 g/g should be the upper limit if carbon conversion is improved by the use of an additional energy source. This assumption was investigated with the methylotrophic yeast Hansenula polymorpha MH 20. Formate served as an additional energy source. The growth yield experiments were performed with a transient-state fermentation technique in which formate was fed via an increasing concentration gradient to a culture growing continuously on glucose. As a result the growth yield on glucose was improve, the extent was dependent on the formate feeding rate, i.e. the slope of this formate gradient. The predicted maximum growth yield of 0.7 g/g was obtained at a slope of the formate gradient of 0.21 g/l·h at a glucose concentration of about 1 g/l. Steeper gradients did not further improve this value, but rather impaired the growth yield due to the appearence of a high residual formate concentration in the fermentation medium. The yield patterns are influenced by the culture pH, a value of at least 4.8 is necessary to achieve the maximum growth yield on glucose. At lower pH formate became increasingly toxic. The ratio of formate to glucose necessary to obtain the maximum yield coefficient was 1...1.6:1 (in grams). On the basis of the energy content of formate a ratio of 1.2...1 (P/O=2) was calculated to substitute the part of glucose which is endoxidized for energy generation. Deviations from this value are explained in terms of the manner of uptake and uncoupling property of formic acid/formate and the existence of a second, formate-“wasting” enzyme.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Mixed substrate utilization ; 14C-Methanol glucose ; Efficiency of methanol dissimilation ; Improvement of growth yield ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Experiments were performed to reveal the extent to which individual heterotrophic substrates of a mixture contribute to the overall carbon and energy metabolism. For this reason Hansenula polymorpha MH 20 was chemostatically (C-limited) cultivated at different growth rates on mixtures of methanol and glucose fed at proportions of 3:1 and 1:3 (in weight units), respectively. The distributions of 14C-carbon from methanol in biomass as well as carbon dioxide (and supernatant) fractions were determined. From these results it followed, firstly, that energy derived from methanol dissimilation was used in part for the incorporation of glucose carbon, resulting in carbon conversion efficiencies for this substrate equivalent to yield coefficients of 0.61–0.69 g/g. Secondly, the growth yield data revealed that the efficiency of methanol conversion had to be increased in order to account for the experimentally determined yield figures. This was further confirmed by theoretical treatment of the growth yield data which showed that these could only be obtained if P/O-quotients for methanol conversion similar to those for glucose, i.e. 2.0–2.5, were considered. The latter property was regarded as the main reason for the observed improvement of growth yield accompanying the simultaneous utilization of methanol and glucose in this yeast.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 144 (1986), S. 62-66 
    ISSN: 1432-072X
    Keywords: Acinetobacter calcoaceticus ; Continuous cultivation ; Acetate growth ; Glucose oxidation ; Energy gain ; Maximum carbon conversion efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Since glucose can be oxidized but not assimilated by Acinetobacter calcoaceticus 69-V the question arose whether energy generated by glucose oxidation can help incorporate carbon from heterotrophic substrates and, if so, what the efficiency of ATP production is like. For this reason this species was grown in the chemostat on acetate. After having reached steady state conditions an increasing concentration of glucose was added. This led to an increase in the biomass level from about 0.4 g/g for growth on acetate alone to 0.6–0.65 g/g in the presence of glucose, independently of either the growth rate or the steepness of the glucose gradient used. This upper value approximates about the limit of the carbon conversion efficiency calculated for non-glycolytic substrates. Glucose was almost exclusively oxidized to gluconic acid, 2- and 5-ketogluconates, and pentose 5-phosphates were found only in traces. These results demonstrate that glucose functions as an additional energy source in Acinetobacter calcoaceticus 69-V. From the transient behaviour of biomass increase and the mixing proportion at which the maximum growth yield on acetate in the presence of glucose was obtained it followed that two mol of ATP must have been generated per mol of glucose oxidized. This property is discussed in terms of coupling glucose dehydrogenase with the respiratory chain.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 87 (1990), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The maximum growth rate of methylotrophic yeasts during growth on methanol is about 0.2 h−1. Since they are able to grow faster on substrates such as glucose we tried to identify the putative limiting step in methanol metabolism within the assimilatory pathway, leading to the formation of a major precursor for biosyntheses, or within the linear dissimilatory sequence. Growth experiments with mixed substrates and determination of some kinetic parameters allowed us to restrict the number of possible pacemaker enzymes. The dissimilatory sequence does not seem to be growth-rate limiting. This also applies to transketolase, transaldolase and fructose-1,6-bisphosphatase. Surprisingly, methanol oxidase appears to be the prime candidate.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 25 (1986), S. 238-244 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Investigations were made into the improvement of growth yield (Y) of Pichia pinus MH 4 growing continuously on methanol by feeding formate so as to create an increasing concentration gradient (transient state). Under particular formate supply conditions, Y could be increased from 0.37 g·g-1 on methanol alone to 0.55 and 0.47 g·g-1 in the presence of formate at dilution rates (D) of 0.045 and 0.075 h-1, respectively. These differences could be explained as being due to a limiting formate consumption rate of 50–60 nmol·min-1·g-1 dry wt., coupled to a net-energy generation independent of D. Any further formate oxidation proceeded without energy gain. Deviations from optimum conditions of biomass increase are discussed in terms of different formate oxidizing systems and uncoupling properties of formate itself. These results are compared to and confirmed by steady-state considerations.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 22 (1985), S. 201-207 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary As the macromolecular composition of microorganisms varies during their life cycle it was asked whether, and to what extent such changes exert any influence on substrate consumption, i.e. growth yield and carbon conversion efficiency, respectively. This question was dealt with in a theoretical study by use of the Y APT max -concept. The growth substrates considered were methanol, acetate and glucose; the latter was assumed to be assimilated via both the glycolytic and the oxidative hexosemonophosphate pathway. Five fictitious biomasses were used which were altered in their proportion of polysaccharides, proteins, lipids, RNA and DNA. As a result, only small variations in the individual “biomass formulae” were obtained. On the basis of the energy balances for the syntheses of all cell constituents it was found that variations in the macromolecular composition of microbial biomass have only a slight effect on carbon conversion efficiency, amounting to maximally 3%. From the material balances it could be calculated that the upper, solely metabolism-determined limit of carbon conversion efficiency is 85% for substrates assimilated glycolytically via phosphoglycerate; for gluconeogenetic substrates, the upper limit was 75%. These limits are essentially determined by carbon loss on the way to amino acid syntheses.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 20 (1984), S. 195-200 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A theoretical consideration is presented of the comparative efficiency of carbon conversion of glucose by the Embden-Meyerhof-Parnas (EMP) and the oxidative hexosemonophosphate (HMP) pathways. As a result it is shown that maximum carbon conversion, that is 89%, is possible when glucose is assimilated via the EMP pathway. This value is diminished in proportion to the participation of the HMP pathway in carbon assimilation and is halved when glucose is incorporated entirely via this pathway. If NADPH is included as a source of energy, glucose may behave both as an excess carbon and an excess energy substrate, the latter being the case when greater portions of the HMP pathway operate, and the extent of this is in turn dependent on the P/O quotient. If NADPH cannot be used for ATP synthesis, glucose remains an excess carbon substrate throughout, although when the HMP pathway accounts for more than 26% of glucose assimilation an increasing excess of reduction equivalents is produced. These results are interpreted in terms of mixed-substrate utilization for improving growth yield when glucose is to be used as the excess carbon component.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1572-9729
    Keywords: Delftia acidovorans MC1 ; 2,4-dichlorophenoxyacetic acid (2,4-D) ; 2-(2,4-dichlorophenoxy) propanoic acid (2,4-DP) ; effect of tfdK gene ; simultaneous utilization of 2,4-D and 2,4-DP ; uptake characteristics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Growth of Delftia acidovorans MC1 on 2,4-dichlorophenoxyacetic acid (2,4-D) and on racemic 2-(2,4-dichlorophenoxy)propanoic acid ((RS)-2,4-DP) was studied in the perspective of an extension of the strain’s degradation capacity at alkaline pH. At pH 6.8 the strain grew on 2,4-D at a maximum rate (μmax) of 0.158 h−1. The half-maximum rate-associated substrate concentration (Ks) was 45 μM. At pH 8.5 μmax was only 0.05 h−1 and the substrate affinity was mucher lower than at pH 6.8. The initial attack of 2,4-D was not the limiting step at pH 8.5 as was seen from high dioxygenase activity in cells grown at this pH. High stationary 2,4-D concentrations and the fact that μmax with dichlorprop was around 0.2 h−1 at both pHs rather pointed at limited 2,4-D uptake at pH 8.5. Introduction of tfdK from D. acidovorans P4a by conjugation, coding for a 2,4-D-specific transporter resulted in improved growth on 2,4-D at pH 8.5 with μmax of 0.147 h−1 and Ks of 267 μM. Experiments with labeled substrates showed significantly enhanced 2,4-D uptake by the transconjugant TK62. This is taken as an indication of expression of the tfdK gene and proper function of the transporter. The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) reduced the influx of 2,4-D. At a concentration of 195 μM 2,4-D, the effect amounted to 90% and 50%, respectively, with TK62 and MC1. Cloning of tfdK also improved the utilization of 2,4-D in the presence of (RS)−2,4-DP. Simultaneous and almost complete degradation of both compounds occurred in TK62 up to D = 0.23 h−1 at pH 6.8 and up to D = 0.2 h−1 at pH 8.5. In contrast, MC1 left 2,4-D largely unutilized even at low dilution rates when growing on herbicide mixtures at pH 8.5.
    Type of Medium: Electronic Resource
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