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Sucrose/D-Fructose/D-Glucose Assay Kit

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00:05  Introduction
00:57  Principle
01:39    Reagent Preparation
03:12  Procedure
08:07  Calculations

Sucrose D-Fructose D-Glucose Assay Kit K-SUFRG Scheme
Product code: K-SUFRG

300 assays (100 of each) per kit

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Content: 300 assays (100 of each) per kit
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 2 years under recommended storage conditions
Analyte: D-Fructose, D-Glucose, Sucrose
Assay Format: Spectrophotometer
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 4 to 80 µg of D-glucose, D-fructose or sucrose per assay
Limit of Detection: 1.38 mg/L
Reaction Time (min): ~ 23 min
Application examples: Beer, fruit juices, soft drinks, milk, jam, honey, dietetic foods, bread, bakery products, dairy products, candies, desserts, confectionery, sweets, ice-cream, fruit and vegetables (e.g. potato), meat products (e.g. sausage), condiments (e.g. ketchup and mustard), feed, tobacco, cosmetics, pharmaceuticals, paper and other materials.
Method recognition: Methods based on this principle have been accepted by NF, EN, NEN, DIN, GOST, IFU, AIJN, MEBAK and IOCCC

The Sucrose/D-Fructose/D-Glucose test kit is suitable for the measurement and analysis of sucrose, D-glucose and D-fructose in plant and food products.

Sucrose, D-glucose and D-fructose are found in most plant and food products. In plant materials, D-glucose and D-fructose occur as free sugars in sucrose, and in a range of oligosaccharides (galactosyl-sucrose oligosaccharides and fructo-oligosaccharides) and polysaccharides such as fructans (inulins), starch, 1,3:1,4-β-D-glucans and cellulose.

Note for Content: The number of manual tests per kit can be doubled if all volumes are halved.  This can be readily accommodated using the MegaQuantTM  Wave Spectrophotometer (D-MQWAVE).

See all of our monosaccharide and disaccharide assay kit products.

Scheme-K-SUFRG SUFRG Megazyme

  • Extended cofactors stability. Dissolved cofactors stable for > 1 year at 4oC.
  • Very competitive price (cost per test) 
  • All reagents stable for > 2 years after preparation 
  • Rapid reaction 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Stabilised D-glucose / D-fructose standard solution included
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance Validation Report
Megazyme publication

Megazyme “advanced” wine test kits general characteristics and validation.

Charnock, S. J., McCleary, B. V., Daverede, C. & Gallant, P. (2006). Reveue des Oenologues, 120, 1-5.

Many of the enzymatic test kits are official methods of prestigious organisations such as the Association of Official Analytical Chemicals (AOAC) and the American Association of Cereal Chemists (AACC) in response to the interest from oenologists. Megazyme decided to use its long history of enzymatic bio-analysis to make a significant contribution to the wine industry, by the development of a range of advanced enzymatic test kits. This task has now been successfully completed through the strategic and comprehensive process of identifying limitations of existing enzymatic bio-analysis test kits where they occurred, and then using advanced techniques, such as molecular biology (photo 1), to rapidly overcome them. Novel test kits have also been developed for analytes of emerging interest to the oenologist, such as yeast available nitrogen (YAN; see pages 2-3 of issue 117 article), or where previously enzymes were simply either not available, or were too expensive to employ, such as for D-mannitol analysis.

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Megazyme publication

Grape and wine analysis: Oenologists to exploit advanced test kits.

Charnock, S. C. & McCleary, B. V. (2005). Revue des Enology, 117, 1-5.

It is without doubt that testing plays a pivotal role throughout the whole of the vinification process. To produce the best possible quality wine and to minimise process problems such as “stuck” fermentation or troublesome infections, it is now recognised that if possible testing should begin prior to harvesting of the grapes and continue through to bottling. Traditional methods of wine analysis are often expensive, time consuming, require either elaborate equipment or specialist expertise and frequently lack accuracy. However, enzymatic bio-analysis enables the accurate measurement of the vast majority of analytes of interest to the wine maker, using just one piece of apparatus, the spectrophotometer (see previous issue No. 116 for a detailed technical review). Grape juice and wine are amenable to enzymatic testing as being liquids they are homogenous, easy to manipulate, and can generally be analysed without any sample preparation.

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Automatic control of chemolithotrophic cultivation of Cupriavidus necator: Optimization of oxygen supply for enhanced bioplastic production.

Lambauer, V., Permann, A., Petrášek, Z., Subotić, V., Hochenauer, C., Kratzer, R. & Reichhartinger, M. (2023). Fermentation, 9(7), 619.

Gas fermentation is an upcoming technology to convert gaseous substrates into value-added products using autotrophic microorganisms. The hydrogen-oxidizing bacteria Cupriavidus necator efficiently uses CO2 as its sole carbon source, H2 as electron donor and O2 as electron acceptor. Surplus CO2 is stored in microbial storage material poly-(R)-3-hydroxybutyrate. O2 supply is the most critical parameter for growth and poly-(R)-3-hydroxybutyrate formation. A narrow O2 optimum between ~0.2 and ~4 mg/L was previously reported. Here, a standard benchtop bioreactor was redesigned for autotrophic growth of C. necator on explosive mixtures of CO2, H2 and O2. The bioreactor was equipped with mass flow control units and O2 and CO2 sensors. A controller for automated gas dosage based on a mathematical model including gas mass transfer, gas consumption and sensor response time was developed. Dissolved O2 concentrations were adjusted with high precision to 1, 2 and 4% O2 saturation (0.4, 0.8 and 1.5 mg/L dissolved O2, respectively). In total, up to 15 g/L cell dry weight were produced. Residual biomass formation was 3.6 ± 0.2 g/L under all three O2 concentrations. However, poly-(R)-3-hydroxybutyrate content was 71, 77 and 58% of the cell dry weight with 1, 2 and 4% dissolved O2, respectively.

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Physicochemical changes during controlled laboratory fermentation of cocoa (CCN-51) with the inclusion of fruits and on-farm inoculation.

Peña González, M. A., Ortiz Urgiles, J. P., Santander Pérez, F. A., Lazo Vélez, M. A. & Caroca Cáceres, R. S. (2023). Brazilian Journal of Food Technology, 26, e2023013.

Fermentation is key to developing the organoleptic characteristics of cocoa beans, as dynamic changes in metabolites have a significant impact on flavors and aromas, hence modifications of this process have been investigated. In this research, the mucilage of CCN-51 cocoa beans was replaced by a mixture of passion fruit (Passiflora edulis) and plantain (Musa paradisiaca L.) pulp, and a controlled fermentation of this mixture was carried out after its spontaneous on-farm inoculation. The physicochemical changes and correlations during the five days of fermentation were evaluated. At the end of the process, the temperature reached 47 ºC in the fermentation mass and pH 5.64 was recorded in the cotyledon. In the first 48 hours, citric acid and fructose were high but at the end of fermentation were 71% and 41.17% lower than at the start of fermentation, respectively. As glucose and fructose were consumed during fermentation, acetic acid and lactic acid levels increased from day two onward, reaching values at the end of the process of 22.48 mg/g and 16.01 mg/g, respectively. In contrast, the bromatological parameters did not show greater variability when comparing each day of fermentation. The data generated and results presented in this study will contribute to the knowledge of possible sensory improvements achieved with the inclusion of pulp fruits in the fermentation stage.

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Genome‐wide transcriptional responses to water deficit during seed development in Pisum sativum, focusing on sugar transport and metabolism.

Morin, A., Porcheron, B., Kodjovi, G. C., Moumen, B., Vriet, C., Maurousset, L., Lemoine, R., Pourtau, N. & Doidy, J. (2023). Physiologia Plantarum, 175(6), e14062.

Agriculture is particularly impacted by global changes, drought being a main limiting factor of crop production. Here, we focus on pea (Pisum sativum), a model legume cultivated for its seed nutritional value. A water deficit (WD) was applied during its early reproductive phase, harvesting plant organs at two key developmental stages, either at the embryonic or the seed-filling stages. We combined phenotypic, physiological and transcriptome analyses to better understand the adaptive response to drought. First, we showed that apical growth arrest is a major phenotypic indicator of water stress. Sugar content was also greatly impacted, especially leaf fructose and starch contents. Our RNA-seq analysis identified 2001 genes regulated by WD in leaf, 3684 genes in root and 2273 genes in embryonic seed, while only 80 genes were regulated during seed-filling. Hence, a large transcriptional reprogramming occurred in response to WD in seeds during early embryonic stage, but no longer during the later stage of nutritional filling. Biological processes involved in transcriptional regulation, carbon transport and metabolism were greatly regulated by WD in both source and sink organs, as illustrated by the expression of genes encoding transcription factors, sugar transporters and enzymes of the starch synthesis pathway. We then looked at the transcriptomic changes during seed development, highlighting a transition from monosaccharide utilization at the embryonic stage to sucrose transport feeding the starch synthesis pathway at the seed-filling stage. Altogether, our study presents an integrative picture of sugar transport and metabolism in response to drought and during seed development at a genome-wide level.

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Land use influences the nutrient concentration and composition of pollen and nectar rewards of wildflowers in human-dominated landscapes.

Pioltelli, E., Guzzetti, L., Larbi, M. O., Celano, R., Piccinelli, A. L., Galimberti, A., Biella, P. & Labra, M. (2024). Science of the Total Environment, 908, 168130.

Plant biodiversity is crucial to satisfy the trophic needs of pollinators, mainly through nectar and pollen rewards. However, a few studies have been directed to ascertain the intraspecific variation of chemical features and the nutritional value of nectar and pollen floral rewards in relation to the alteration of landscapes due to human activities. In this study, by using an existing scenario of land use gradients as an open air laboratory, we tested the variation in pollen and nectar nutrient profiles along gradients of urbanization and agriculture intensity, by focusing on sugar, aminoacids of nectar and phytochemicals of pollen from local wild plants. We also highlighted bioactive compounds from plants primary and secondary metabolism due to their importance for insect wellbeing and pollinator health. We surveyed 7 different meadow species foraged by pollinators and common in the main land uses studied. The results indicated that significant variations of nutritional components occur in relation to different land uses, and specifically that the agricultural intensification decreases the sugars and increases the antioxidant content of flower rewards, while the urbanization is positively associated with the total flavonoid content in pollen. These effects are more evident in some species than in others, such as Lotus corniculatus L. (Fabaceae) and Malva sylvestris L. (Malvaceae), as shown by the untargeted metabolomic investigation. This study is crucial for understanding the nutritional landscape quality for pollinators in association to different land uses and sets a base for landscape management and planning of pollinator-friendly strategies by improving the quality of plant rewards to provide benefits to pollinator health in various environmental contexts.

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Multiscale variability in nutrients and secondary metabolites in a bat‐dispersed neotropical fruit.

Gelambi, M. & Whitehead, S. R. (2023). Ecology and Evolution, 13(9), e10453.

Ripe fleshy fruits contain not only nutrients but also a diverse array of secondary metabolites. Nutrients serve as a reward for mutualists, whereas defensive metabolites protect the fruit against pests and predators. The composition of these chemical traits is highly variable, both across different plants and even within repeating structures on the same individual plant. This intraspecific and intraindividual variation has important fitness consequences for both plants and animals, yet patterns of variation and covariation in nutrients and secondary metabolites are not well understood, especially at smaller scales. Here, we investigate the multiscale variation and covariation between nutrients and defensive metabolites in Piper sancti-felicis ripe fruits. Means and measures of variation of sugars, proteins, phenolics, and alkenylphenols vary greatly among plants, and at least 50% of the trait variation occurs at the intraindividual level. Also, we found that proteins, but not sugars, were correlated with phenolics and alkenylphenols at multiple scales, suggesting trait variation in protein content may be more constrained than sugars. Our findings emphasize the importance of examining patterns across scales and provide the groundwork to better understand how complex patterns of variation and covariation in nutrients and defensive metabolites shape ecological interactions surrounding fruits.

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Selenium-Fortified Kombucha-Pollen Beverage by In Situ Biosynthesized Selenium Nanoparticles with High Biocompatibility and Antioxidant Activity.

Tritean, N., Dima, Ș. O., Trică, B., Stoica, R., Ghiurea, M., Moraru, I., Cimpean, A., Oancea, F. & Constantinescu-Aruxandei, D. (2023). Antioxidants, 12(9), 1711.

Biogenic selenium nanoparticles (SeNPs) have been shown to exhibit increased bioavailability. Fermentation of pollen by a symbiotic culture of bacteria and yeasts (SCOBY/Kombucha) leads to the release of pollen content and enhances the prebiotic and probiotic effects of Kombucha. The aim of this study was to fortify Kombucha beverage with SeNPs formed in situ by Kombucha fermentation with pollen. Response Surface Methodology (RSM) was used to optimize the biosynthesis of SeNPs and the pollen-fermented Kombucha beverage. SeNPs were characterized by Transmission electron microscopy energy-dispersive X-ray spectroscopy (TEM-EDX), Fourier-transform infrared spectroscopy (FTIR), Dynamic light scattering (DLS), and Zeta potential. The pollen-fermented Kombucha beverage enriched with SeNPs was characterized by measuring the total phenolic content, antioxidant activity, soluble silicon, saccharides, lactic acid, and the total content of Se0. The polyphenols were identified by liquid chromatography–mass spectrometry (LC-MS). The pollen and the bacterial (nano)cellulose were characterized by scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX), FTIR, and X-Ray diffraction (XRD). We also assessed the in vitro biocompatibility in terms of gingival fibroblast viability and proliferation, as well as the antioxidant activity of SeNPs and the pollen-fermented Kombucha beverage enriched with SeNPs. The results highlight their increased biological performance in this regard.

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First attempt to find spectral patterns in cork oak leaves revealing the interaction of the plant with Phytophthora cinnamomi.

Guerra, R., Pires, R., Brázio, A., Cavaco, A. M., Schütz, G., & Coelho, A. C. (2023), In Press.

Cork oak tree (Quercus suber L.) symbolises Montado landscape in Portugal and is a central element in the country's social and economic history. In recent decades, the loss of thousands of cork oaks has been reported, revealing the ongoing decline of these agro-forestry ecosystems. This emblematic tree of the Mediterranean Basin is host to the soil-born root pathogen Phytophthora cinnamomi, an active cork oak decline driver. In this framework, the early diagnosis of trees infected by the oomycete by non-invasive methods should contribute to the sustainable management of cork oak ecosystems and motivated this work. Using leaf reflectance spectroscopy it was possible to distinguish plants inoculated with P. cinnamomi from non-inoculated plants in a long course assay and to argue about the factors contributing to this differentiation. Spectral acquisitions in the visible-near infrared (400–1100 nm) were performed at 63, 78, 91, 126 and 248 days after inoculation (DAI), on the abaxial and adaxial faces of the leaves, and the spectral data was arranged in a three-way array of samples × wavelengths × days. The separation of the two plant groups were attributed to variations in water content (4v (O−H)) and/or a shift in the red edge and to structural modifications in the epidermal layer and/or the mesophyll of the leaves observed in the visible range. These spectral signatures can assist in the field identification of cork oaks that are interacting with P. cinnamomi.

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Physiological and sucrose metabolic responses to waterlogging stress in balloon flower (Platycodon grandiflorus (Jacq.) A. DC).

Ji, H. S. & Hyun, T. K. (2023). Physiology and Molecular Biology of Plants, 1-10.

Waterlogging stress is a major limiting factor resulting in stunted growth and loss of crop productivity, especially for root crops. However, physiological responses to waterlogging have been studied in only a few plant models. To gain insight into how balloon flower (Platycodon grandiflorus (Jacq.) A. DC) responds to waterlogging stress, we investigate changes to sucrose metabolism combined with a physiological analysis. Although waterlogging stress decreased the photosynthetic rate in balloon flower, leaves exhibited an increase in glucose (ninefold), fructose (4.7-fold), and sucrose (2.1-fold), indicating inhibition of sugar transport via the phloem. In addition, roots showed a typical response to hypoxia, such as the accumulation of proline (4.5-fold higher than in control roots) and soluble sugars (2.1-fold higher than in control roots). The activities and expression patterns of sucrose catabolizing enzymes suggest that waterlogging stress leads to a shift in the pathway of sucrose degradation from invertase to sucrose synthase (Susy), which consumes less ATP. Furthermore, we suggest that the waterlogging-stress-induced gene PlgSusy1 encodes the functional Susy enzyme, which may contribute to improving tolerance in balloon flower to waterlogging. As a first step toward understanding the waterlogging-induced regulatory mechanisms in balloon flower, we provide a solid foundation for further understanding waterlogging-induced alterations in source-sink relationships.

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Physiochemical Changes of European Pear cv. Conference and Asian Pear cv. Yali during Cold Storage.

Híc, P., Kožíšková, J., Omastová, P., Balík, J., Goliáš, J. & Horák, M. (2023). Horticulturae, 9(3), 378.

This study evaluated the physiochemical changes of one commercial European pear variety ‘Conference’ and one Asian pear variety ‘Yali’ during 60 days of storage at 1°C. Content of organic acids, ethylene and formation of CO2 were determined using HPLC/UV-VIS and GC/FID and TCD detectors, respectively. This study presents an evaluation of the influence of the above-mentioned factors and long-term storage on quality parameters, including the antioxidant capacity of an important European and Asian pear cultivar. There was a significant effect of temperature on respiration rate in both varieties. Development of the respiration intensity had a similar course in European and Asian pears. The high values of CO2 production at the beginning of storage for the variety ‘Conference’ (14.08 mg·kg−1·h−1) and ‘Yali’ (23.37 mg·kg−1·h−1) were followed by a sharp decline of 80% and 83% at later stages of storage at 1°C, respectively. Ethylene formation in fruit increased with storage time and was the highest after 60 days in cold storage, especially in ‘Yali’ (7.14 µL·kg−1·h−1). The results show the relation between ethylene formation and ripening-related fruit parameters. The ‘Yali’ variety showed negligible changes in soluble solids content and flesh firmness during storage ranging from 0.35 to 0.60 MPa. The most represented sugar determined enzymatically was fructose and malic acid was the dominant organic acid in pear fruit. Antioxidant activity determined using the FRAP and DPPH methods did not change significantly during 60 days of storage.

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Upcycling of black currant pomace for the production of a fermented beverage with Wolfiporia cocos.

Sommer, S., Hoffmann, J. L., Fraatz, M. A. & Zorn, H. (2023). Journal of Food Science and Technology, 60(4), 1313-1322.

Pomace as a side stream from black currant juice production is mostly discarded, even though it is rich in nutrients like protein, fiber, sugars, anthocyanins, polyphenols, and other secondary metabolites. Fungi from the division of Basidiomycota have a great enzymatic toolbox to recycle these complex mixtures of nutrients. In particular, the edible medicinal fungus Wolfiporia cocos has been described as a suitable biocatalyst to form pleasant aroma compounds in fermentation processes. Therefore, medium optimization, upscaling, and filtration were performed to produce a beverage based on black currant pomace fermented with W. cocos. A trained panel described the beverage as highly pleasant, reminiscent of honey, flowers and berries with a well-balanced sour and sweet taste. The flavor compounds linalool (citrus), geraniol (flowery), phenylacetic acid (honey), methyl phenylacetate (honey), eugenol (clove), and 2-phenylethanol (rose) were produced during fermentation and the concentrations exceeded their respective odor thresholds. The produced beverage was evaluated with 8.0 ± 1.4 from 10 for the question of whether panelists would buy the product. Fungal fermentation with the edible fungus W. cocos enabled the production of a highly pleasant beverage and additionally may reduce waste by using pomace and table sugar as sole ingredients.

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Effect of pulsed electric field pre-treatment on the debittering process of cherry kernels.

Giancaterino, M., Fauster, T., Krottenthaler, A. & Jäger, H. (2022). Innovative Food Science & Emerging Technologies, 103234.

Cherry kernels occur in significant amounts as waste material during the processing of fruits. However, their subsequent use is limited due to the presence of cyanogenic glycosides, which are potentially dangerous to human health. In this study, the application of pulsed electric fields (PEF) was investigated as pre-treatment to improve the debittering process and to facilitate the degradation of cyanide precursors, naturally present in cherry kernels. Diverse PEF treatments were carried out at constant electric field strength of E = 2.2 kV/cm and specific energy input was varied between 10 and 50 kJ/kg, varying the number of pulses. Two different debittering procedures were performed with a common incubation time 0-20 h at 40°C: a) incubation of whole kernels in deionized water; b) incubation of whole kernels without water stored in air at 80% relative humidity. HPLC analysis was used to examine the kinetics of the amygdalin and HCN contents. In both debittering methods, the PEF-treated samples with the highest intensity (2.2 kV/cm, 50 kJ/kg) showed higher and faster detoxification efficiency for the investigated compounds as compared to the untreated sample. In particular, the PEF treated samples incubated with water showed a reduction in the amygdalin and HCN contents of up to 86% (up to 72% of the raw material content.). Moreover, the PEF pre-treatment led to comparable efficiency in amygdalin reduction in both debittering processes: 86% reduction for the incubation with water and 81% for the incubation without water. Consequently, the combined application of PEF and the debittering process including incubation without water has remarkable potential as an industrial application due to its inherent reduced water consumption, and therefore, diminished wastewater management issues. A further advantage of this process is the minimizing of sugar loss typically occurring during the debittering through soaking.

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The Effect of Dekkera bruxellensis Concentration and Inoculation Time on Biochemical Changes and Cellulose Biosynthesis by Komagataeibacter intermedius.

Devanthi, P. V. P., Pratama, F., Kho, K., Taherzadeh, M. J. & Aslanzadeh, S. (2022). Journal of Fungi, 8(11), 1206.

Bacterial Cellulose (BC) is a biopolymer with numerous applications. The growth of BC-producing bacteria, Komagataeibacter intermedius, could be stimulated by Dekkera bruxellensis, however, the effect on BC yield needs further investigation. This study investigates BC production and biochemical changes in the K. intermedius-D. bruxellensis co-culture system. D. bruxellensis was introduced at various concentrations (103 and 106 CFU/mL) and inoculation times (days 0 and 3). BC yield was ~24% lower when D. bruxellensis was added at 103 CFU/mL compared to K. intermedius alone (0.63 ± 0.11 g/L). The lowest BC yield was observed when 103 CFU/mL yeast was added on day 0, which could be compromised by higher gluconic acid production (10.08 g/L). In contrast, BC yields increased by ~88% when 106 CFU/mL D. bruxellensis was added, regardless of inoculation time. High BC yield might correlate with faster sugar consumption or increased ethanol production when 106 CFU/mL D. bruxellensis was added on day 0. These results suggest that cell concentration and inoculation time have crucial impacts on species interactions in the co-culture system and product yield.

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Use of apple pomace, glycerine, and potato wastewater for the production of propionic acid and vitamin B12.

Piwowarek, K., Lipińska, E., Hać-Szymańczuk, E., Kolotylo, V. & Kieliszek, M. (2022). Applied Microbiology and Biotechnology, 106(17), 5433-5448.

Propionic acid bacteria (PAB) are a source of valuable metabolites, including propionic acid and vitamin B12. Propionic acid, a food preservative, is synthesized from petroleum refining by-products, giving rise to ecological concerns. Due to changing food trends, the demand for vitamin B12 has been expected to increase in the future. Therefore, it is necessary to look for new, alternative methods of obtaining these compounds. This study was conducted with an aim of optimizing the production of PAB metabolites using only residues (apple pomace, waste glycerine, and potato wastewater), without any enzymatic or chemical pretreatment and enrichment. Media consisting of one, two, or three industrial side-streams were used for the production of PAB metabolites. The highest production of propionic acid was observed in the medium containing all three residues (8.15 g/L, yield: 0.48 g/g). In the same medium, the highest production of acetic acid was found - 2.31 g/L (0.13 g/g). The presence of waste glycerine in the media had a positive effect on the efficiency of propionic acid production and P/A ratio. The concentration of vitamin B12 obtained in the wet biomass of Propionibacterium freudenreichii DSM 20271 ranged from 90 to 290 µg/100 g. The highest production of cobalamin was achieved in potato wastewater and apple pomace, which may be a source of the precursors of vitamin B12 - cobalt and riboflavin. The results obtained show both propionic acid and vitamin B12 can be produced in a more sustainable manner through the fermentation of residues which are often not properly managed.

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Safety Information
Symbol : GHS05, GHS08
Signal Word : Danger
Hazard Statements : H314, H360
Precautionary Statements : P201, P202, P260, P264, P280
Safety Data Sheet
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