Formation Of Vitamin B12 Research Articles

Obtaining Novel Vitamin B12 Production Strains Acetobacter malorum HFD 3141 and Acetobacter orientalis HFD 3031 from Home-Fermented Sourdough

Vitamin B12 is a critical nutrient in vegan and vegetarian lifestyles as plant-based vitamin sources are rare. Traditional fermented foods could be enriched by adding vitamin B12-producing bacteria to offer non-animal vitamin sources. The aim was to isolate a vitamin B12 producer that is capable of producing the human-active vitamin even at low pH values so that it can be used in fruit juice fortification. Therefore, fermented foods (homemade and industrial) and probiotics were screened for vitamin B12 production strains. A modified microbiological vitamin B12 assay based on Lactobacillus delbrueckii subsp. lactis DSM 20355 was used to identify vitamin B12-containing samples and the presence of vitamin B12-producing strains. The screening resulted in isolating several positive strains for vitamin B12 formation derived from sourdough. Mass spectrometry confirmed the biosynthesis of solely the human physiologically active form. Species identification carried out by the German Strain Collection of Microorganisms and Cell Cultures resulted in two species: Acetobacter orientalis and Acetobacter malorum, of which two isolates were further characterised. The potential for cobalamin biosynthesises in food matrixes was demonstrated for A. malorum HFD 3141 and A. orientalis HFD 3031 in apple juice at different pH values (2.85–3.80). The isolates synthesised up to 18.89 µg/L and 7.97 µg/L vitamin B12 at pH 3.80. The results of this study suggest that acetic acid bacteria (AAB) and fermented acetic acid foods are promising resources for vitamin B12 and its producers, which might have been overlooked in the past.

Prevalence of Cobalt in the Environment and Its Role in Biological Processes.

Cobalt (Co) is an essential trace element for humans and other animals, but high doses can be harmful to human health. It is present in some foods such as green vegetables, various spices, meat, milk products, seafood, and eggs, and in drinking water. Co is necessary for the metabolism of human beings and animals due to its key role in the formation of vitamin B12, also known as cobalamin, the biological reservoir of Co. In high concentrations, Co may cause some health issues such as vomiting, nausea, diarrhea, bleeding, low blood pressure, heart diseases, thyroid damage, hair loss, bone defects, and the inhibition of some enzyme activities. Conversely, Co deficiency can lead to anorexia, chronic swelling, and detrimental anemia. Co nanoparticles have different and various biomedical applications thanks to their antioxidant, antimicrobial, anticancer, and antidiabetic properties. In addition, Co and cobalt oxide nanoparticles can be used in lithium-ion batteries, as a catalyst, a carrier for targeted drug delivery, a gas sensor, an electronic thin film, and in energy storage. Accumulation of Co in agriculture and humans, due to natural and anthropogenic factors, represents a global problem affecting water quality and human and animal health. Besides the common chelating agents used for Co intoxication, phytoremediation is an interesting environmental technology for cleaning up soil contaminated with Co. The occurrence of Co in the environment is discussed and its involvement in biological processes is underlined. Toxicological aspects related to Co are also examined in this review.

Dietary Utilization Drives the Differentiation of Gut Bacterial Communities between Specialist and Generalist Drosophilid Flies.

ABSTRACTGut bacteria play vital roles in the dietary detoxification, digestion, and nutrient supplementation of hosts during dietary specialization. The roles of gut bacteria in the host can be unveiled by comparing communities of specialist and generalist bacterial species. However, these species usually have a long evolutionary history, making it difficult to determine whether bacterial community differentiation is due to host dietary adaptation or phylogenetic divergence. In this regard, we investigated the bacterial communities from two Araceae-feeding Colocasiomyia species and further performed a meta-analysis by incorporating the published data from Drosophila bacterial community studies. The compositional and functional differentiation of bacterial communities was uncovered by comparing three (Araceae-feeding, mycophagous, and cactophilic) specialists with generalist flies. The compositional differentiation showed that Bacteroidetes and Firmicutes inhabited specialists, while more Proteobacteria lived in generalists. The functional prediction based on the bacterial community compositions suggested that amino acid metabolism and energy metabolism are overrepresented pathways in specialists and generalists, respectively. The differences were mainly associated with the higher utilization of structural complex carbohydrates, protein utilization, vitamin B12 acquisition, and demand for detoxification in specialists than in generalists. The complementary roles of bacteria reveal a connection between gut bacterial communities and fly dietary specialization.IMPORTANCE Gut bacteria may play roles in the dietary utilization of hosts, especially in specialist animals, during long-term host-microbe interaction. By comparing the gut bacterial communities between specialist and generalist drosophilid flies, we found that specialists harbor more bacteria linked to complex carbohydrate degradation, amino acid metabolism, vitamin B12 formation, and detoxification than do generalists. This study reveals the roles of gut bacteria in drosophilid species in dietary utilization.

Karakterisasi kandungan vitamin B12, folat dan isoflavon tempe kedelai dengan isolat murni Rhizopus oryzae, Rhizopus oligosporus, dan Rhizopus stolonifer sebagai bahan pangan fungsional

Tempeh is a fermented food ingredient from soybeans that uses various Rhizopus molds, such as Rhizopus oligosporus, Rhizopus oryzae, Rhizophus stolonifer. In this study, the preparation of tempe using mixed culture was carried out in the ratio of Rhizopus oryzae, Rhizopus oligosporus, Rhizopus stolonifer inoculums, namely 1: 1: 1; 1: 2: 1; 2: 1: 2; 1: 1: 2; 2: 1: 1; market yeast "Raprima", produces the characteristics of tempe according to SNI 01-3144-2009, formation of Vitamin B12, Folate and Isoflavones as well as pH, and Total Bacteria. The content of vitamin B12 was obtained in yeast "raprima" of 0.013 g / 50g of tempeh while tempeh made with pure inoculums of Rhizopus sp did not contain vitamin B12 or folic acid, pH value of 4.2 and total bacteria 14.3x108 cfu / ml and organoleptic reception. Tempe 1: 2: 1 has the characteristics of color, aroma, taste, texture and viscosity acceptable to the panelists.

Deficiencia de cobalto em Bovinos: Revisión

Cobalt (Co) is a relatively rare mineral and an essential element necessary for the formation of vitamin B12 (cobalamin) in humans and animals. Cobalt deficiency is a disease that only affects ruminants that ingest in their diet low concentrations of this mineral, which leads to the deficiency of this central vitamin in several metabolic pathways in the animals' organisms. The objectives of this work were to make a brief review of cobalt deficiency in ruminants, addressing epidemiological aspects, clinical and pathological characteristics, diagnosis, treatment, and control of this disease. Cobalt deficiency is clinically characterized by inappetence, weight loss, and reproductive problems. The disease occurs in all biomes and regions of Brazil, affecting mainly animals raised in extensive management. The presumptive diagnosis is based on the history and observation of the herds, in which the animals lose weight despite the abundant pasture and the control is made by supplementation with the mineral.

Relationship between Vitamin B12 and Cobalt Metabolism in Domestic Ruminant: An Update.

Simple SummaryWe review the role of cobalt and vitamin B12 in animals, especially in ruminants. Vitamin B12 is an essential part of the enzyme systems involved in multiple metabolic reactions and mainly in the formation of energy from ruminal fermentation. Signs of deficiency, as well as cobalt toxicity, in animals are described. The level of cobalt in ruminants can be assessed by measuring the blood or tissue concentrations of cobalt or vitamin B12, as well as the level of some metabolites such as malonate, homocysteine or transobolamine in blood or methylmalonic acid in urine. The requirement for cobalt (Co) is around 0.11 ppm (mg/kg) dry matter (DM) in the diet, although current recommendations advise supplementing the diet up to 0.20 mg Co/kg DM, which seems to increase animal production, especially in dairy cattle.Cobalt, as a trace element, is essential for rumen microorganisms for the formation of vitamin B12. In the metabolism of mammals, vitamin B12 is an essential part of two enzymatic systems involved in multiple metabolic reactions, such as in the metabolism of carbohydrates, lipids, some amino acids and DNA. Adenosylcobalamin and methylcobalamin are coenzymes of methylmalonyl coenzyme A (CoA) mutase and methionine synthetase and are essential for obtaining energy through ruminal metabolism. Signs of cobalt deficiency range from hyporexia, reduced growth and weight loss to liver steatosis, anemia, impaired immune function, impaired reproductive function and even death. Cobalt status in ruminant animals can be assessed by direct measurement of blood or tissue concentrations of cobalt or vitamin B12, as well as the level of methylmalonic acid, homocysteine or transcobalamin in blood; methylmalonic acid in urine; some variables hematological; food consumption or growth of animals. In general, it is assumed that the requirement for cobalt (Co) is expressed around 0.11 ppm (mg/kg) in the dry matter (DM) diet; current recommendations seem to advise increasing Co supplementation and placing it around 0.20 mg Co/kg DM. Although there is no unanimous criterion about milk production, fattening or reproductive rates in response to increased supplementation with Co, in some investigations, when the total Co of the diet was approximately 1 to 1.3 ppm (mg/kg), maximum responses were observed in the milk production.

Cobalt distribution in the soils of São Miguel Island (Azores): From volcanoes to health effects

Volcanic soils cover only approximately 1% of the Earth's surface, yet they support 10% of the world's population due to their inherent fertility. However, deep magmatic processes may lead to depletion of certain elements in volcanic rocks. The imbalance of essential elements, such as cobalt, in soil parent materials can affect the health of plants and grazing animals and, thus, humans. Within the particular geological context of the Azores, the present study aims to assess cobalt concentration in volcanic soils to predict the risk of cobalt deficiency in animals and humans.Samples from agricultural topsoils and pasture grass were collected in six volcanic regions of São Miguel Island, and their physicochemical properties were measured, including selected transition metal element contents, such as iron, manganese and cobalt. The soil cobalt concentration was below 5 mg/kg in Povoação< Furnas/Congro< Sete Cidades< Fogo and was higher than 10 mg/kg in Picos< Nordeste. Cobalt concentrations were very low in pasture grass. Values below 0.1 mg/kg were observed in Furnas/Congro<Povoação<Fogo<Picos<Sete Cidades, and only the volcanic region of Nordeste had adequate values of cobalt (0.34 ± 0.19 mg/kg) in pasture grass to fulfill animal needs. The concentrations of iron and manganese were also significantly higher in the two latter volcanic regions. These differences result from the pedogenesis of volcanic rocks with distinct geochemical compositions related to different degrees of magmatic evolution.The results show that there is a significant lack of cobalt in agricultural soils on São Miguel and that it is important to predict the risk of cobalt deprivation due to its significant biological value. Soils deficient in cobalt can reduce plant growth, result in the impairment of vitamin B12 formation in animals, and consequently affect animal and human health.

3-Aminotriazole protects from CoCl2-induced ototoxicity by inhibiting the generation of reactive oxygen species and proinflammatory cytokines in mice.

Cobalt is an essential heavy metal that is necessary for the formation of vitamin B12 (hydroxocobalamin). However, exposure to excess cobalt for a prolonged period can harm the human body, causing pulmonary fibrosis, blindness, deafness, and peripheral neuropathy. 3-Aminotriazole (3-AT) is a catalase inhibitor that is often used to investigate the physiological effects of catalase. The present study found that injection of 3-AT in mice significantly reduced CoCl2-induced hearing impairment. In cultured organ of Corti explants from rats, 3-AT treatment protected hair cells from CoCl2-induced cytotoxicity. To determine the mechanism by which 3-AT protected from CoCl2-induced ototoxicity, we used the HEI-OC1 auditory cell line. Pretreatment with 10 mM 3-AT attenuated CoCl2-induced accumulation of ROS and induction of proinflammatory cytokine expression. Interestingly, these protective effects of 3-AT did not require catalase activity, as demonstrated by a series of experiments using RNA interference-mediated catalase knockdown in HEI-OC1 cells and using catalase-deficient mouse embryonic fibroblasts. Our results demonstrated the mechanisms of CoCl2-induced ototoxicity that may provide better ways to prevent the ototoxic effect of cobalt exposure.

Roles of cobalt in biosynthesis stimulation of a cytotoxic compound from marine-derived Aspergillus glaucus

Effects of trace elements on aspergiolide A biosynthesis by marine-derived filamentous fungus Aspergillus glaucus HB 1–19 were investigated. Cobalt (Co2+) and nickel (Ni2+) stimulated aspergiolide A biosynthesis while zinc (Zn2+) inhibited it. Equal cobalt (Co2+) addition at 48, 72 and 96h with total amount of 0.052mM most effectively enhanced aspergiolide A production. The time courses indicated that mycelia growth, sugar utilization and aspergiolide A biosynthesis were enhanced by Co2+. Finally, feeding 0.052mM Co2+ increased mycelia growth, sugar utilization and aspergiolide A production by 12.2%, 13.7% and 46.7%, respectively. The results of organic acids analysis indicated that extracellular pyruvate increased while fumarate decreased in Co2+ addition cultures. Moreover, feeding vitamin B12, which combined cobalt to be a cofactor form, increased aspergiolide A production by 59.1%. This indicated Co2+ functioned by facilitating pyruvate accumulation and increasing vitamin B12 formation that were beneficial to aspergiolide A biosynthesis that is via polyketide pathway.

Study on methane fermentation and production of vitamin B12 from alcohol waste slurry.

We studied biogas fermentation from alcohol waste fluid to evaluate the anaerobic digestion process and the production of vitamin B12 as a byproduct. Anaerobic digestion using acclimated methanogens was performed using the continuously stirred tank reactor (CSTR) and fixed-bed reactor packed with rock wool as carrier material at 55 degrees C. We also studied the effects of metal ions added to the culture broth on methane and vitamin B12 formation. Vitamin B12 production was 2.92 mg/L in the broth of the fixed-bed reactor, twice that of the CSTR. The optimum concentrations of trace metal ions added to the culture liquid for methane and vitamin B12 production were 1.0 and 8 mL/L for the CSTR and fixed-bed reactor, respectively. Furthermore, an effective method for extracting and purifying vitamin B12 from digested fluid was developed.

Cobalt.

Cobalt is a relatively rare magnetic element with properties similar to iron and nickel. The two valance states are cobaltous (II) and cobaltic (III) and the former is the most common valance used in the chemical industry. Cobalt occurs in nature primarily as arsenides, oxides, and sulfides. Most of the production of cobalt involves the metallic form used in the formation of cobalt superalloys. The term "hard metal" refers to compounds containing tungsten carbide (80-95%) combined with matrices formed from cobalt (5-20%) and nickel (0-5%). For the general population, the diet is the main source of exposure to cobalt. In the occupational setting, exposure to cobalt alone occurs primarily during the production of cobalt powders. In other industrial exposures (e.g., hard metal, diamond polishing), additional agents (tungsten) modulate the toxicity of cobalt. Cobalt is an essential element necessary for the formation of vitamin B12 (hydroxocobalamin); however, excessive administration of this trace element produces goiter and reduced thyroid activity. In 1966, the syndrome "beer drinker's cardiomyopathy" appeared in Quebec City, Canada, and was characterized by pericardial effusion, elevated hemoglobin concentrations, and congestive heart failure. An interstitial pulmonary fibrosis has been associated with industrial exposure to hard metal dust (tungsten and cobalt), but not to cobalt alone. Exposure to cobalt alone produces an allergic contact dermatitis and occupational asthma. Treatment of cobalt toxicity is primarily supportive.

Biosynthesis of vitamin B12 in anaerobic bacteria--experiments with Eubacterium limosum on the transformation of 5-hydroxy-6-methyl-benzimidazole, its nucleoside, its cobamide, and of 5-hydroxybenzimidazolylcobamide in vitamin B12.

In anaerobic bacteria 5-hydroxybenzimidazole and 5-hydroxy-6-methylbenzimidazole are precursors of the 5,6-dimethylbenzimidazole moiety of vitamin B12. In order to elucidate the pathway from these bases to vitamin B12, experiments on the transformation of 5-hydroxy-6-methylbenzimidazole, of 5-hydroxy-6-methylbenzimidazole-alpha-D-ribofuranoside, of 5-hydroxybenzimidazolylcobamide and of 5-hydroxy-6-methylbenzimidazolylcobamide into vitamin B12 were carried out. The vitamin B12 synthesized by the anaerobe Eubacterium limosum in the presence of 5-hydroxy-6-methylbenzimidazole and L-[methyl-13C]methionine was subjected to NMR spectroscopy. It revealed that the methyl group at C5 of the 5,6-dimethylbenzimidazole moiety was 13C labeled, whereas the methyl group at C6 was unlabeled. This shows that the transformation of 5-hydroxy-6-methylbenzimidazole into the base moiety of vitamin B12 occurs regiospecifically. 5-Hydroxy-6-methylbenzimidazole-alpha-D-ribofuranoside as well as 5-hydroxybenzimidazolylcobamide and 5-hydroxy-6-methylbenzimidazolylcobamide were also transformed into vitamin B12 by E. limosum. When 5-hydroxy-6-methylbenzimidazolylcobamide 13C labeled at C2 of the base part and 14C labeled in the ribose was used for this experiment, the vitamin B12 obtained from this cobamide was 13C and 14C labeled in the same positions. This demonstrates that the alpha-glycosidic bond of the precursor cobamide is not split during the formation of vitamin B12. It can be deduced from these results that the precursor bases are transformed regiospecifically into their alpha-nucleotides, and partially into their cobamides. The alpha-nucleotides are then transformed into alpha-ribazole-5'-phosphate and, subsequently, into vitamin B12. Most likely the cobamides are degraded to the alpha-nucleotides before being used for the biosynthesis of vitamin B12. A pathway for the latter process is suggested.

The Crucial Contribution of Starved Resting Cells to the Elucidation of the Pathway of Vitamin B12 Biosynthesis

Resting cells were crucial in elucidating the steps of vitamin B12 biosynthesis, and their use led to the following conclusions: (1) 5-aminolevulinic acid is a common precursor of vitamin B12 and porphyrins in various groups of microorganisms. (2) Glutamine is the donor of the amide groups in the biosynthesis of the amidated corrin structure of vitamin B12. (3) Methylated derivatives of uroporphyrinogen III, called corriphyrins, are precursors of the corrin ring of vitamin B12. They are formed in the initial steps of uroporphyrinogen III transformation to cobyrinic acid. (4) The initial common steps of heme and vitamin B12 synthesis are controlled by heme, while the biosynthetic pathway starting with methylation of uroporphyrinogen III and leading to corrinoids is controlled by vitamin B12. Aeration interferes with vitamin B12 formation but not that of porphyrins. The mechanism is that of oxygen suppression of methylation of uroporphyrinogen III. These conclusions were completely supported by subsequent studies of others. This shows that suspensions of resting microbial cells can be profitably employed in solving important problems of microbial biosynthesis of physiologically active compounds.

Vitamin B12 production by Citrobacter freundii or Klebsiella pneumoniae during tempeh fermentation and proof of enterotoxin absence by PCR.

The influence of some fermentation parameters on vitamin B12 formation by strains of Citrobacter freundii and Klebsiella pneumoniae isolated from Indonesian tempeh samples during tempeh fermentation was investigated. A decrease in fermentation temperature from 32 to 24 degrees C led to a decrease in vitamin B12 formation. Inoculation of soybeans with different numbers of cells of C. freundii at the beginning of solid-substrate fermentation showed that only the velocity of vitamin formation and not the final amount of vitamin formed depended on the number of cells. The addition of cobalt and 5,6-dimethylbenzimidazole increased the vitamin B12 content of tempeh. Nevertheless, levels of incorporation of the two precursors into the vitamin B12 molecule were very low. Neither C. freundii nor K. pneumoniae possessed the genes encoding the enterotoxins Shiga-like toxin SLT IIA, heat-labile enterotoxin LT Ih, and heat-stable enterotoxin ST Ih, as indicated by PCR. This result supports the suggested use of these two strains to form vitamin B12 during tempeh fermentation in Indonesia.

Formation of vitamins by pure cultures of tempe moulds and bacteria during the tempe solid substrate fermentation.

The formation of water soluble vitamins (vitamin B12, vitamin B6, riboflavin, thiamine, nicotinic acid and nicotinamide) during the tempe solid substrate fermentation was investigated. The role of several strains of Rhizopus oligosporus, R. arrhizus, and R. stolonifer and the role of several bacteria in the vitamin formation process were checked. All fungal and bacterial strains were isolated from Indonesian tempe and soaking water samples. The Rhizopus strains formed riboflavin, nicotinic acid, nicotinamide and vitamin B6. The final concentrations of these substances depended on the different strains involved and on the fermentation time. Isolates of R. oligosporus were generally the best vitamin formers. The moulds did not produce physiologically active vitamin B12. The thiamine content decreased during fermentation. The addition of bacteria, which had been selected in a screening for vitamin B12 production, resulted in an increase of physiologically active vitamin B12. Citrobacter freundii and Klebsiella pneumoniae showed the best formation capabilities. Furthermore, the bacteria produced riboflavin and vitamin B6 in addition to the moulds. The influence of Rhizopus on the vitamin B12 formation of Cit. freundii was also investigated. The vitamin content of tempe that was fermented with the mould and the bacterium was three times as high as a control fermentation with Cit. freundii only.

Stimulation of vitamin B12 formation in aerobically-grownRhodopseudomonas gelatinosa under microaerobic condition

Photopigments and vitamin B12 formation ofRhodopseudomonas gelatinosa were enhanced by a stepwise change of the culture condition from aerobic (oxidation-reduction potential, ORP>+110 mV) to microaerobic condition (ORP=0 to −200 mV). During the microaerobic culture in the malateglutamate medium, δ-aminolevulinic acid synthetase (ALAS) increased 2 to 4 folds in 4 h with the increases in intracellular content of carotenoid, bacteriochlorophyll and vitamin B12. Effects of light illumination on vitamin B12 formation could not be observed. Further, the production of SCP enriched with vitamin B12 and photopigments from cassava starch was done by changing aerobic to microaerobic culture resulting that intracellular carotenoid, bacteriochlorophyll and vitamin B12 increased to 310, 960 and 38 μg/g cell from 230, 0 and 25 μg/g cell of aerobic culture, respectively.

Isolation of a formate-assimilating bacterium and its vitamin B12 formation.

Isolation of a formate-assimilating bacterium and its vitamin B12 formation.

Vitamin B12 formation from various actinomycetes isolated from bulgarian soils

240 зтаммов актиномицетов, выделенных из ризосферы различных растений и из различных почв, исследовались на наличие витамина В12 путем культивирования в жидкой питательной среде. 73% из исследованных штаммов обладают способностью образовывать витамин В12, в количестван порядка 0,0025–1,6 μ/мл.

Microbiological formation of vitamin B12. II. Synthesis of vitamin B12-active substances by bacteria isolated from clams.

Vitamin B122occurred in certain seaweeds and this vitamin and other cobalamins were demonstrated regularly in marine invertebrates. A Bacillus and a Micrococcus isolated from fresh clams were selected for detailed study on the basis of their pronounced ability to form cobalamins, and because they appeared to be typical of the bacterial flora of these invertebrates. The Bacillus formed vitamin B12and other cobalamins, and this activity was stimulated by addition of Factor B and of certain purines or benziminazoles to the medium. With both organisms, 6-nitrobenziminazole proved particularly effective in stimulating formation of both vitamin B12and of other cobalamins, and Factor B stimulated this activity. Formation of cobalamins was associated with a marked decrease in Factor B. The cobalamins were associated with the bacterial cells, and no cobalamin synthesis by washed cell suspensions in phosphate buffer was observed.

BIOSYNTHESIS OF VITAMIN B12 FROM CO60-LABELED COBALT-PORPHYRIN BY AGED CELL HOMOGENATE OF FUGLENA GRACILIS

1. Biosynthesis of vitamin B12 by aged cell homogenate of B12-nonproducing microbe was tested, and it was found by bioautography that the in vitro formation of vitamin B12 from cobalt-porphyrin and 5, 6-dimethylbenzimidazole was always positive.2. Further evidence was obtained by using Co60-labeled Co-porphyrin, and biosynthesis of Co60-labeled vitamin B12 was confirmed by radioautography.3. From these findings it is concluded that cobalt-porphyrin may be available for the formation of vitamin B12 in vitro by the intracellular enzyme of an Euglena strain which is ordinarily an nonproducer of vitamin B12.