Analysis of vitamin B<sub>12</sub> in seawater and marine sediment porewater using ELISA

Zhu, Qingzhi; Aller, Robert C.; Kaushik, Aleya

doi:10.4319/lom.2011.9.515

Cited by 15 publications

(14 citation statements)

References 30 publications

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“…Vitamin B 12 was quantified by Enzyme-Linked Immunosorbent Assay, ELISA (Immunolab GmbH, B12-E01. Kassel, Germany) according to Zhu et al (2011). These authors tested the ELISA with seawater samples and found close to 100 percent recovery of cyanocobalamin, methylcobalamin, hydroxycobalamin and coenzyme B 12 , but the method cannot distinguish between these different forms of B 12 .…”

Section: Visual Observationsmentioning

confidence: 99%

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

et al. 2018

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Recent research has shown that in aquatic systems pairs of prokaryote and eukaryote species exercise symbiotic exchanges of metabolites that are essential for the proliferation of either species. Using dinoflagellate Lingulodinium polyedrum cultures and a factorial design, we examined its growth at different concentrations of vitamin B 1 (thiamine) and B 12 (cobalamin). When both vitamins were at their lowest concentrations tested, 0.033 pM of B 1 and 0.053 pM of B 12 the growth was limited. When axenic L. polyedrum was co-cultured with the bacterium Dinoroseobacter shibae, a known B 1 and B 12 producer, then L. polyedrum grew at the same rate as in culture media supplemented with B 1 and B 12. In the L. polyedrum vitamin-limited culture (V-L), the abundance of attached and free-living D. shibae was higher than in the vitamin-replete (V-R) culture. In the V-R and V-L co-cultures the measured particulate B 12 (pB 12) concentration of attached and free-living D. shibae were in the range of 4.7 × 10 −19 to 3 × 10 −18 and 8.4 × 10 −21 to1.2 × 10 −19 (mol cell −1), respectively. Without B 12 or B 7 (biotin) added to the culture medium of a co-culture of L. polyedrum and D. shibae, the measured dissolved B 12 (dB 12) concentration was more than 60 pM higher than necessary for unlimited growth rates of L. polyedrum. In the same culture we measured B 7 in the L. polyedrum particulate fraction (pB 7 ; 4.7 × 10 −19 to 9.4 × 10 −19 mol cell −1). We suggest that in response to the production of B 1 and B 12 by D. shibae to supply L. polyedrum requeriments, the latter produced B 7 , which is required by D. shibae, and in our culture was only produced by L. polyedrum when D. shibae was present. We propose that D. shibae can control L. polyedrum through the release of B 1 and B 12 , and L. polyedrum can control D. shibae through the release of B 7. D. shibae is also auxotroph for niacin and 4-amino-benzoic acid, not provided by the culture medium. Therefore, L. polyedrum might affect a similar control through the release of these specific compounds and organic substrate necessary for the growth of D. shibae.

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Section: Visual Observationsmentioning

confidence: 99%

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

et al. 2018

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“…The column was then extracted with 5 ml methanol (≥99.9%; Sigma-Aldrich #34860). To prepare the samples for the ELISA immunoassay, the methanol was evaporated in a water bath at 60°C, applying a gentle vacuum for 6 min; the dried samples were re-suspended in distilled water (2 ml final volume) (Zhu et al 2011) to be used for the ELISA assay. The extraction efficiency was tested using the B 12 standard of the ELISA kit, by processing the standard the same way as the samples including solid phase extraction, methanol evaporation including heating, and taking up in water.…”

Section: Extraction and Elisa Quantification Of Dissolved And Intracementioning

confidence: 99%

“…A sensitive, specific ELISA (Immunolab) was used to quantify B 12 (Zhu et al 2011). This method does not distinguish between the different B 12 components Zhu et al (2011) could quantitatively detect (cyanocobalamin, methylcobalamin, coenzyme B 12 , and hy -dro xycobalamin) and compared favorably with the HPLC method. They also showed that the method does not recognize other B vitamins including thiamin (B 1 ), riboflavin (B 2 ), and pyridoxine (B 6 ).…”

Section: Extraction and Elisa Quantification Of Dissolved And Intracementioning

confidence: 99%

“…They also showed that the method does not recognize other B vitamins including thiamin (B 1 ), riboflavin (B 2 ), and pyridoxine (B 6 ). Zhu et al (2011) did not test the response of the assay to pseudocobalamin (a vitamin that can serve a similar metabolic function as B 12 ); therefore, we cannot be sure that this method included the quantification of pseudocobalamin. Thus, when we refer to our data as B 12 this includes the above-mentioned different forms of the cobalamin.…”

Section: Extraction and Elisa Quantification Of Dissolved And Intracementioning

confidence: 99%

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B12 production by marine microbial communities and Dinoroseobacter shibae continuous cultures under different growth and respiration rates

Villegas-Mendoza¹,

Cajal-Medrano²,

Maske³

2019

Aquat. Microb. Ecol.

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In situ dissolved B 12 concentration in marine ecosystems is controlled by the balance between rates of release of B 12 by prokaryotes, uptake by prokaryotes and eukaryotes, and abiotic degradation. We used chemostats at a range of specific growth rates (μ, d −1 ; 0.1 to 1) with natural communities of prokaryotes and monospecific cultures of a B 12 producer, Dinoroseobacter shibae. We measured the dissolved B 12 concentration produced in the culture (B 12-d), the B 12 in the particulate fraction (B 12-p), cell concentration, respiration rate, particulate organic carbon and nitrogen (POC, PON), and the 16S amplicon composition. Total dissolved B 12 concentrations (0.92 to 4.90 pmol l −1) were comparable to those found in the surface ocean. B 12-p concentration was 6 to 35 times higher than B 12-d. B 12-d , B 12-p , and community composition showed no relation to μ for either natural populations or D. shibae. The chemostats allowed calculation of the rates of production: B 12-d (0.34 ± 0.28 pmol l −1 d −1) and B 12-p (5.65 ± 2.34 pmol l −1 d −1), and the B 12 cell quota (900 to 3300 molecules cell −1). In multispecies and D. shibae cultures, B 12 production rates per cell increased with respiration rates (volumetric or per cell), and with rates of cellular organic carbon and nitrogen production. Rates increased with μ, but not the concentrations of B 12-d or of B 12-p. To understand the physiological and ecological dynamics of B 12 , concentrations alone are insufficient since they do not provide rates, which are important in understanding the dynamics between producers and consumers.

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“…Indeed, microbially produced cobalt-binding ligands are found in seawater (Saito and Moffett, 2001;Saito et al, 2005) and have also been shown to be produced in sulphidogenic bioreactors (Jansen et al, 2005). It is also likely that some fraction of total dissolved cobalt in methane seeps is present as B12; sediment porewater B12 concentrations are 10-50 pM in sulphidic coastal sediments (Zhu et al, 2011), more abundant than free divalent Co 2+ , which is ∼0.01 pM at > 1 mM HS − in Hydrate Ridge porewaters assuming speciation follows the dynamics shown in Fig. 3.…”

Section: Cobalt Porewater Speciation Bioavailability and Microbial Tmentioning

confidence: 99%

Geochemical, metagenomic and metaproteomic insights into trace metal utilization by methane‐oxidizing microbial consortia in sulphidic marine sediments

Glass

Steele

et al. 2013

Environmental Microbiology

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Summary Microbes have obligate requirements for trace metals in metalloenzymes that catalyse important biogeochemical reactions. In anoxic methane‐ and sulphide‐rich environments, microbes may have unique adaptations for metal acquisition and utilization because of decreased bioavailability as a result of metal sulphide precipitation. However, micronutrient cycling is largely unexplored in cold (≤ 10°C) and sulphidic (> 1 mM ΣH2S) deep‐sea methane seep ecosystems. We investigated trace metal geochemistry and microbial metal utilization in methane seeps offshore Oregon and California, USA, and report dissolved concentrations of nickel (0.5–270 nM), cobalt (0.5–6 nM), molybdenum (10–5600 nM) and tungsten (0.3–8 nM) in Hydrate Ridge sediment porewaters. Despite low levels of cobalt and tungsten, metagenomic and metaproteomic data suggest that microbial consortia catalysing anaerobic oxidation of methane (AOM) utilize both scarce micronutrients in addition to nickel and molybdenum. Genetic machinery for cobalt‐containing vitamin B12 biosynthesis was present in both anaerobic methanotrophic archaea (ANME) and sulphate‐reducing bacteria. Proteins affiliated with the tungsten‐containing form of formylmethanofuran dehydrogenase were expressed in ANME from two seep ecosystems, the first evidence for expression of a tungstoenzyme in psychrophilic microorganisms. Overall, our data suggest that AOM consortia use specialized biochemical strategies to overcome the challenges of metal availability in sulphidic environments.

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Analysis of vitamin B₁₂ in seawater and marine sediment porewater using ELISA

Cited by 15 publications

References 30 publications

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

B12 production by marine microbial communities and Dinoroseobacter shibae continuous cultures under different growth and respiration rates

Geochemical, metagenomic and metaproteomic insights into trace metal utilization by methane‐oxidizing microbial consortia in sulphidic marine sediments

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Analysis of vitamin B12 in seawater and marine sediment porewater using ELISA

Cited by 15 publications

References 30 publications

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

The B-Vitamin Mutualism Between the Dinoflagellate Lingulodinium polyedrum and the Bacterium Dinoroseobacter shibae

B12 production by marine microbial communities and Dinoroseobacter shibae continuous cultures under different growth and respiration rates

Geochemical, metagenomic and metaproteomic insights into trace metal utilization by methane‐oxidizing microbial consortia in sulphidic marine sediments

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Analysis of vitamin B₁₂ in seawater and marine sediment porewater using ELISA