Kinetic evaluation and monitoring of methanogen culture based upon fluorometry

Taya, Masahito; Aoki, Nobuhiro; Kobayashi, Takeshi

doi:10.1007/bf00257030

Cited by 16 publications

(8 citation statements)

References 20 publications

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“…Speaking of the specific CoF420 content on the biomass unit, its time coincidence with the biomass curve is observed only in the logarithmic growth phase. This result coincides with the data obtained for Methanobrevibacter arbon'philus and Methanobacterium themoautotrophicum [4]. The relation of the ascents of the straight lines I and 111, and I1 and IV represented in Fig.…”

Section: Resultssupporting

confidence: 81%

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Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Kuschk

Kevbrin

1989

Acta Biotechnologica

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Fed-batch cultivation of Methanobacterium formieicum 15-281 was performed under substrate limiting conditions. Specific rate of culture growth amounted to 0.041 h-l, cell yield related to substrate was 0.6 mg proteine/mM formate. Significant increasc of coenzyme F,, content in cells depending on time of cultivation was observed. Increase of the fluorescence intensity of the medium was also observed with the accumulation of biomuss. These parameters were suggested for tracking culture growth. The possibility of application of fluorescent factors for methanogenic population control in the anaerobic digester is discussed. IntrodiictionMethane-forming bacteria are the last step i n a chain of organic waste decomposition occuring in the digester. To estimate methanogelis i n the consortium quantitatively, it is necessary t o find a method for evaluating either methanogenic biomass directly, or a parameter connected with it. Methanogenic bacteria differ from other members of the consortium by possessing coenzyme F,,, (CoF42,) which becomes fluorescent after oxidizing [l]. The structure of CoF,,, was determined and its biological function was partially clarified [2]. The method of biomass analysis on CoF420 content is relatively simple [3,4]. Of all methanogens species growing on H, + CO, mixture and on formate contain the greatest quantity of CoF,,, [5]. Among such species there are many representatives of the genus Methanobacterium, to which belongs formate-using species Methanobacterium formicicum. The strains JF-1 [6], MF [7] are well studied from the point of view of the physiology of growth. We report here about the investigation results of Methanobacterium formicicum strain 2-281. Materials and Methods Organisms and cultivation medium : Methunobacterium formicicum 2-281 (DSM 3722) was received from T. N. ZH~LINA [8]. Mineral medium contained the following components (in g/l) : NH,Cl -0.33; KH,PO, -0.33; MgCl, x 6H,O -0.33; CaCl, x 2H20 -0.33; KCl -0.33; HCOONa -5.0; Na,S x 9H,O -0.4. To the medium were also

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Section: Resultssupporting

confidence: 81%

“…The structure of CoF,,, was determined and its biological function was partially clarified [2]. The method of biomass analysis on CoF420 content is relatively simple [3,4]. Of all methanogens species growing on H, + CO, mixture and on formate contain the greatest quantity of CoF,,, [5].…”

Section: Introdiictionmentioning

confidence: 99%

Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Kuschk

Kevbrin

1989

Acta Biotechnologica

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“…Methanobacteriumn thernoaulltotrophic(um, Methanobreli-baceter arboriphiluis (31), M. bryantii, M. bacrker-i FUSARO (15), and Methanobacteriurn fornicicuin cultivated in continuous culture (5). The intracellular coenzyme F420 content of M. rnazei (230 nmol g of protein-'), M. barkeri MS cultured on methanol (474 nmol g of protein-'), and M. barker-i MS cultured on H2-CO (530 nmol g of protein-') agree well with previous values ( Table 1).…”

Section: Discussionmentioning

confidence: 99%

Changes in concentrations of coenzyme F420 analogs during batch growth of Methanosarcina barkeri and Methanosarcina mazei

Peck

1989

Appl Environ Microbiol

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Coenzyme F420 has been assayed by high-performance liquid chromatography with fluorimetric detection; this permits quantification of individual coenzyme F420 analogs whilst avoiding the inclusion of interfering material. The total intracellular coenzyme F420 content of Methanosarcina barkeri MS cultivated on methanol and on H2-C02 and of Methanosarcina mazei S-6 cultured on methanol remained relatively constant during batch growth. The most abundant analogs in M. barkeri were coenzymes F420-2 and F420-4, whilst in M. mazei coenzymes F420-2 and F420-3 predominated. Significant changes in the relative proportions of the coenzyme F420 analogs were noted during batch growth, with coenzymes F420-2 and F420-4 showing opposite responses to each other and the same being also true for coenzymes F420-3 and F420-5. This suggests that an enzyme responsible for transferring pairs of glutamic acid residues may be active. The degradation fragment FO was also detected in cells in late exponential and stationary phase. Coenzyme F420 analogs were present in the culture supernatant of both methanogens, in similar proportions to that in the cells, except for FO which was principally located in the supernatant.

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“…Only few data on comparable studies in pure cultures of methanogens are available. Taya et al (29) showed that culture fluorescence paralleled cell concentration for Methanobacterium thermoautotrophicum and Methanobrevibacter arboriphilus. However, only relative fluorescence was determined and not absolute amounts of F420.…”

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confidence: 99%

Relationship of Intracellular Coenzyme F ₄₂₀ Content to Growth and Metabolic Activity of Methanobacterium bryantii and Methanosarcina barkeri

Heine-Dobbernack¹,

Schoberth²,

Sahm³

1988

Appl Environ Microbiol

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The use of F420 as a parameter for growth or metabolic activity of methanogenic bacteria was investigated. Two representative species of methanogens were grown in batch culture: Methanobacterium bryantii (strain M.o.H.G.) on H2 and CO2, and Methanosarcina barkeri (strain Fusaro) on methanol or acetate. The total intracellular content of coenzyme F420 was followed by high-resolution fluorescence spectroscopy. F420 concentration in M. bryantii ranged from 1.84 to 3.65 ,Lmol g of protein-'; and in M. barkeri grown with methanol it ranged from 0.84 to 1.54 ,umol g-depending on growth conditions. The content of F420 in M. barkeri was influenced by a factor of 2 depending on the composition of the medium (minimal or complex) and by a factor of 3 to 4 depending on whether methanol or acetate was used as the carbon source. A comparison of F420 content with protein, cell dry weight, optical density, and specific methane production rate showed that the intracellular content of F420 approximately followed the increase in biomass in both strains. In contrast, no correlation was found between specific methane production rate and intracellular F420 content. However, qCH4(F420), calculated by dividing the methane production rate by the coenzyme F420 concentration, almost paralleled qCH4(protein). These results suggest that F420 may be used as a specific parameter for estimating the biomass, but not the metabolic activity, of methanogens; hence qCH4(F420) determined in mixed populations with complex carbon substrates must be considered as measure of the actual methanogenic activity and not as a measure of potential activity.

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Kinetic evaluation and monitoring of methanogen culture based upon fluorometry

Cited by 16 publications

References 20 publications

Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Changes in concentrations of coenzyme F420 analogs during batch growth of Methanosarcina barkeri and Methanosarcina mazei

Relationship of Intracellular Coenzyme F ₄₂₀ Content to Growth and Metabolic Activity of Methanobacterium bryantii and Methanosarcina barkeri

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Kinetic evaluation and monitoring of methanogen culture based upon fluorometry

Cited by 16 publications

References 20 publications

Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Fed‐batch cultivation of methanobacterium formicicum and its fluorometric monitoring

Changes in concentrations of coenzyme F420 analogs during batch growth of Methanosarcina barkeri and Methanosarcina mazei

Relationship of Intracellular Coenzyme F 420 Content to Growth and Metabolic Activity of Methanobacterium bryantii and Methanosarcina barkeri

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Relationship of Intracellular Coenzyme F ₄₂₀ Content to Growth and Metabolic Activity of Methanobacterium bryantii and Methanosarcina barkeri