Medium optimization for a methanol utilizing bacterium based on chemostat theory

Tsuchiya, Yoshinobu; Nishio, Naomichi; Nagai, Shiro

doi:10.1007/bf00503507

Cited by 11 publications

(6 citation statements)

References 4 publications

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“…We concluded from these experiments that the growth rate slightly increases with MgCl 2 concentrations up to 0.125 mM, plateaus, and then begins to decrease at concentrations above 1.5 mM. This result is consistent with a previous study that found MgCl 2 limited the growth of AM1 on methanol at concentrations below 0.121 mM [38]. For this reason, we set the MgCl 2 level at 0.5 mM in MP medium, a level on the plateau between the beneficial and detrimental effects, and kept the concentration of all other medium components at the midpoint of their tested levels in MP medium.…”

Section: Methods and Resultssupporting

confidence: 92%

Development of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquens

et al. 2013

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Methylobacterium extorquens strains are the best-studied methylotrophic model system, and their metabolism of single carbon compounds has been studied for over 50 years. Here we develop a new system for high-throughput batch culture of M. extorquens in microtiter plates by jointly optimizing the properties of the organism, the growth media and the culturing system. After removing cellulose synthase genes in M. extorquens strains AM1 and PA1 to prevent biofilm formation, we found that currently available lab automation equipment, integrated and managed by open source software, makes possible reliable estimates of the exponential growth rate. Using this system, we developed an optimized growth medium for M. extorquens using response surface methodologies. We found that media that used EDTA as a metal chelator inhibited growth and led to inconsistent culture conditions. In contrast, the new medium we developed with a PIPES buffer and metals chelated by citrate allowed for fast and more consistent growth rates. This new Methylobacterium PIPES (‘MP’) medium was also robust to large deviations in its component ingredients which avoided batch effects from experiments that used media prepared at different times. MP medium allows for faster and more consistent growth than other media used for M. extorquens.

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Section: Methods and Resultssupporting

confidence: 92%

Development of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquens

et al. 2013

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“…The basal culture medium used is a synthetic mineral medium supplemented with 0.5% (v/v) methanol described by Tsuchiya et al (1980) and modified as follows (g/l): methanol, 5; (NH4)2SO4, 1; KH2PO4, 1.3; Na2HPO4, 2.13; MgSO4"7H20, 0.45; CaC12.2H20, 0.003; FeSO4"7H20, 0.001; and trace elements such as Mn 2+, Zn 2+, Cu 2+, Mo z+, Co z+ and Na +. The medium was adjusted to pH 7.0 with 2 M NaOH solution.…”

Section: Methodsmentioning

confidence: 99%

Partial purification and characterization of ?-hydroxybutyric acid dehydrogenase of a methylotrophic bacterium, Pseudomonas 135

Daniel

Barbotin

Kim

et al. 1992

Appl Microbiol Biotechnol

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An intracellular enzyme, D(-)-fl-hydroxybutyric acid dehydrogenase involved in an intracellular poly-D(-)-fl-hydroxybutyric acid degradation was isolated from a facultative methylotrophic bacterium, P s e u d o m o n a s 135, grown on methanol as a sole carbon and energy source. This enzyme was partially purified to l l.6-fold by ammonium sulphate fractionation and a dye-affinity chromatography. The enzyme catalysed simultaneously the oxidation of D(-)-fl-hydroxybutyric acid (Dfl-HB) and the reduction of acetoacetate. The optimum pH was 8.5 for the oxidation reaction and 5.5-6.0 for the reduction reaction, and the enzyme was stable for 2 weeks at -20 ° C. The Km values for oxidation and reduction reactions were determined as 1.84 mM for Dfl-HB, 0.244 mM for NAD +, 0.319 mM for acetoacetate and 0.032 mM for NADH, respectively. It was also found that D-lactate and NADH significantly inhibited the oxidation reaction by competitive inhibition, and acetoacetate by non-competitive inhibition, respectively. The inhibition constants were determined as 1.49 mM for D-lactate, 0.196mM for NADH and 1.82mM for acetoacetate, respectively. According to an experiment with resting cells, it seemed that the enzyme was constitutive.

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“…The levels of both elements should slightly exceed a given amount of carbon source to favour growth and protein production and to prevent accumulation of storage substances such as polysaccharides (Haggstrrm 1977;Litchfield 1979;Tsuchiya et al 1980;Goldberg and Er-el 1981;Tempest and Wouters 1981). However, excess phosphorus in the medium produced intracellular polyphosphate granules (Drozd and Linton 1981).…”

Section: Discussionmentioning

confidence: 99%

Chemostat optimization of biomass production of a mixed bacterial culture utilizing methanol

Aburuwaida

Banat

Hamdan

1990

Appl Microbiol Biotechnol

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A continuous culture technique was used to optimize the medium composition and growth conditions of a mixed bacterial culture utilizing methanol. The improved medium resuited in satisfactory growth, high-yield coefficients and gave a product containing reduced polysaccharide concentrations. Optimal growth and biomass yields occurred at pH 6.8 a temperature of 37°C and dissolved oxygen at >20% saturation. The maximum growth rate was 0.58 h -1 and maximum biomass yield 0.48 g g-1. The protein content of the product ranged between 81%-83%, and nucleic acid content between 10%-12%, increasing with growth rate. The amino acid profile of the mixed culture product m e t and, in some cases, exceeded the UN Food and Agricultural Organization standard, indicating a good source of feed protein.

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Medium optimization for a methanol utilizing bacterium based on chemostat theory

Cited by 11 publications

References 4 publications

Development of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquens

Development of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquens

Partial purification and characterization of ?-hydroxybutyric acid dehydrogenase of a methylotrophic bacterium, Pseudomonas 135

Chemostat optimization of biomass production of a mixed bacterial culture utilizing methanol

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