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L-Lactic Acid (L-Lactate) Assay Kit

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L-Lactic Acid L-Lactate Assay Kit K-LATE Scheme
Product code: K-LATE

50 assays (manual) / 500 assays (microplate) / 450 assays (auto-analyser)

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Content: 50 assays (manual) / 500 assays (microplate) / 450 assays (auto-analyser)
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: L-Lactic Acid
Assay Format: Spectrophotometer, Microplate, Auto-analyser
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Increase
Linear Range: 0.3 to 30 µg of L-lactic acid per assay
Limit of Detection: 0.21 mg/L
Reaction Time (min): ~ 10 min
Application examples: Wine, beer, soft drinks, milk, dairy products (e.g. cream, milk / whey powder, cheese, condensed milk and yogurt), foods containing milk (e.g. dietetic foods, bakery products, baby food, chocolate, sweets and ice-cream), egg, egg products (e.g. egg powder), baking additives, vinegar, fruit and vegetables, processed fruit and vegetables (e.g. tomatoes), meat products, food additives, feed, paper (and cardboard), cosmetics, pharmaceuticals and other materials (e.g. biological cultures, samples, etc.).
Method recognition: Methods based on this principle have been accepted by DIN, GOST, IDF, EEC, EN, ISO, OIV, IFU, AIJN and MEBAK

The L-Lactic Acid (L-Lactate) Assay Kit is used for the specific measurement and analysis of L-lactic acid (L-lactate) in beverages, meat, dairy and food products.

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).

Explore our complete list of organic acid assay kits.

Scheme-K-LATE LATE 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 
  • Standard included
  • Suitable for manual, microplate and auto-analyser formats
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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Chemical Composition of Sour Beer Resulting from Supplementation the Fermentation Medium with Magnesium and Zinc Ions.

Ciosek, A., Fulara, K., Hrabia, O., Satora, P. & Poreda, A. (2020). Biomolecules, 10(12), 1599.

The bioavailability of minerals, such as zinc and magnesium, has a significant impact on the fermentation process. These metal ions are known to influence the growth and metabolic activity of yeast, but there are few reports on their effects on lactic acid bacteria (LAB) metabolism during sour brewing. This study aimed to evaluate the influence of magnesium and zinc ions on the metabolism of Lactobacillus brevis WLP672 during the fermentation of brewers’ wort. We carried out lactic acid fermentations using wort with different mineral compositions: without supplementation; supplemented with magnesium at 60 mg/L and 120 mg/L; and supplemented with zinc at 0.4 mg/L and 2 mg/L. The concentration of organic acids, pH of the wort and carbohydrate use was determined during fermentation, while aroma compounds, real extract and ethanol were measured after the mixed fermentation. The addition of magnesium ions resulted in the pH of the fermenting wort decreasing more quickly, an increase in the level of L-lactic acid (after 48 h of fermentation) and increased concentrations of some volatile compounds. While zinc supplementation had a negative impact on the L. brevis strain, resulting in a decrease in the L-lactic acid content and a higher pH in the beer. We conclude that zinc supplementation is not recommended in sour beer production using L. brevis WLP672.

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Single-use printed biosensor for l-lactate and its application in bioprocess monitoring.

Theuer, L., Randek, J., Junne, S., Neubauer, P., Mandenius, C. F. & Beni, V. (2020). Processes, 8(3), 321.

There is a profound need in bioprocess manufacturing for low-cost single-use sensors that allow timely monitoring of critical product and production attributes. One such opportunity is screen-printed enzyme-based electrochemical sensors, which have the potential to enable low-cost online and/or off-line monitoring of specific parameters in bioprocesses. In this study, such a single-use electrochemical biosensor for lactate monitoring is designed and evaluated. Several aspects of its fabrication and use are addressed, including enzyme immobilization, stability, shelf-life and reproducibility. Applicability of the biosensor to off-line monitoring of bioprocesses was shown by testing in two common industrial bioprocesses in which lactate is a critical quality attribute (Corynebacterium fermentation and mammalian Chinese hamster ovary (CHO) cell cultivation). The specific response to lactate of the screen-printed biosensor was characterized by amperometric measurements. The usability of the sensor at typical industrial culture conditions was favorably evaluated and benchmarked with commonly used standard methods (HPLC and enzymatic kits). The single-use biosensor allowed fast and accurate detection of lactate in prediluted culture media used in industrial practice. The design and fabrication of the biosensor could most likely be adapted to several other critical bioprocess analytes using other specific enzymes. This makes this single-use screen-printed biosensor concept a potentially interesting and versatile tool for further applications in bioprocess monitoring.

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Milk fermented with Lactobacillus rhamnosus R0011 induces a regulatory cytokine profile in LPS-challenged U937 and THP-1 macrophages.

Jeffrey, M. P., Taggart, H. J., Strap, J. L., Edun, G. & Green-Johnson, J. M. (2020). Current Research in Food Science, 3, 51-58.

Fermented dairy products have become attractive functional foods for the delivery of probiotics and their biologically active metabolites. The aim of this study was to examine the immunomodulatory activity of milk fermented with the probiotic lactic acid bacterium Lactobacillus rhamnosus R0011 (LrF) on macrophages challenged with lipopolysaccharide (LPS), a potent pro-inflammatory stimulus. To this end, human THP-1 or U937 monocytes were differentiated into resting macrophages then stimulated with LPS and co-incubated with the LrF or with milk controls. Levels of pro-inflammatory and immunoregulatory cytokines were determined by enzyme-linked immunosorbent assays. Culturing of LPS-stimulated U937 macrophages with either the whole or filtered LrF resulted in an increase in Interleukin (IL)-1Ra production relative to the negative control. THP-1 macrophages cultured with the LrF demonstrated an increase in LPS-induced IL-10 and IL-1β production, while production of LPS-induced IL-6, sCD54, IL-8, IL-1β, TNF-α, IL-12p70 and Transforming Growth Factor-β (TGF-β) was unaffected. Further, the LrF induced the expression of DC-SIGN and CD206, markers of immunoregulatory M2 macrophage polarization, in LPS-challenged THP-1 macrophages. Taken together, milk fermented with L. rhamnosus R0011 increased regulatory cytokine production from LPS-challenged U937 and THP-1 macrophages, while simultaneously up-regulating the production of IL-1β and expression of DC-SIGN and CD206, a profile characteristic of polarization into the immunoregulatory M2 macrophage phenotype.

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ZnO nanowire-based fluorometric enzymatic assays for lactate and cholesterol.

Briones, M., Busó-Rogero, C., Catalán-Gómez, S., García-Mendiola, T., Pariente, F., Redondo-Cubero, A. & Lorenzo, M. E. (2020). Microchimica Acta, 187(3), 1-9.

A rapid fluorometric method is described for the determination of lactate and cholesterol by using ZnO nanowires (ZnO NWs). The assay is based on the detection of the hydrogen peroxide generated during the enzymatic reactions of the oxidation of lactate or cholesterol. Taking advantage of the electrostatic interactions between the enzymes and the ZnO NWs, two bioconjugates were prepared by mixing the nanomaterial and the enzymes, viz. lactate oxidase (LOx) or cholesterol oxidase (ChOx). The enzymatically generated hydrogen peroxide quenches the fluorescence of the ZnO NWs, which have emission peaks at 384 nm and at 520 nm under 330 nm photoexcitation. H2O2 quenches the 520 nm band more strongly. Response is linear up to 1.9 μM lactate concentration, and up to 1.1 μM cholesterol concentration. Relative standard deviation was found to be 5%. The detection limits for lactate and cholesterol are 0.54 and 0.24 μM, respectively.

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Pediococcus spp.-fermented chicken meat for dogs.

Lee, E., Nam, K. T., Lee, K. W. & Lee, S. R. (2020). Journal of Animal Science and Technology, 62(1), 84.

An experiment was conducted to evaluate Pediococcus spp.-fermented chicken meat as a snack for dogs. The fermented or non-fermented snacks used in this study were prepared through the following process; meat mixtures containing 52.8% MDCM, 35.2% chicken breast meat (CBM) and 9.7% corn starch were inoculated with or without Pediococcus spp., incubated at 37°C for 24 h and then sterilized at 121°C for 20 min. During the 24-h fermentation, the pH of fermented chicken snack dropped rapidly with concomitant increase in number of lactic acid bacteria. The nutritional composition was not altered by fermentation. In vitro pepsin nitrogen digestibility was higher (p < 0.05) in the fermented snack compared with the non-fermented snack. Upon storage at room temperature for 14 days, bacteria grew slowly in fermented vs. non-fermented snack samples. In a palatability trial, dogs preferred non-fermented over fermented snack food. In 12-d-long feeding trial, fecal ammonia content was lowered, but fecal lactic acid content was increased in dogs fed the fermented vs. non-fermented snack food. Our study shows that the fermented MDCM-based snack exhibited good preservability upon storage, and improved in vitro nitrogen digestibility and fecal characteristics in dogs.

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Immobilization of Lactobacillus rhamnosus in polyvinyl alcohol/calcium alginate matrix for production of lactic acid.

Radosavljević, M., Lević, S., Belović, M., Pejin, J., Djukić-Vuković, A., Mojović, L. & Nedović, V. (2020). Bioprocess and Biosystems Engineering, 43(2), 315-322.

Immobilization of Lactobacillus rhamnosus ATCC7469 in poly(vinyl alcohol)/calcium alginate (PVA/Ca-alginate) matrix using “freezing–thawing” technique for application in lactic acid (LA) fermentation was studied in this paper. PVA/Ca-alginate beads were made from sterile and non-sterile PVA and sodium alginate solutions. According to mechanical properties, the PVA/Ca-alginate beads expressed a strong elastic character. Obtained PVA/Ca-alginate beads were further applied in batch and repeated batch LA fermentations. Regarding cell viability, L. rhamnosus cells survived well rather sharp immobilization procedure and significant cell proliferation was observed in further fermentation studies achieving high cell viability (up to 10.7 log CFU g−1) in sterile beads. In batch LA fermentation, the immobilized biocatalyst was superior to free cell fermentation system (by 37.1%), while the highest LA yield and volumetric productivity of 97.6% and 0.8 g L−1 h−1, respectively, were attained in repeated batch fermentation. During seven consecutive batch fermentations, the biocatalyst showed high mechanical and operational stability reaching an overall productivity of 0.78 g L−1 h−1. This study suggested that the “freezing–thawing” technique can be successfully used for immobilization of L. rhamnosus in PVA/Ca-alginate matrix without loss of either viability or LA fermentation capability.

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Exploiting Day-and Night-Time Metabolism of Synechocystis sp. PCC 6803 for Fitness-Coupled Fumarate Production around the Clock.

Du, W., Jongbloets, J. A., Guillaume, M., van de Putte, B., Battaglino, B., Hellingwerf, K. J.  & Branco dos Santos, F. (2019). ACS Synthetic Biology, 8(10), 2263-2269.

Cyanobacterial cell factories are widely researched for the sustainable production of compounds directly from CO2. Their application, however, has been limited for two reasons. First, traditional approaches have been shown to lead to unstable cell factories that lose their production capability when scaled to industrial levels. Second, the alternative approaches developed so far are mostly limited to growing conditions, which are not always the case in industry, where nongrowth periods tend to occur (e.g., darkness). We tackled both by generalizing the concept of growth-coupled production to fitness coupling. The feasibility of this new approach is demonstrated for the production of fumarate by constructing the first stable dual-strategy cell factory. We exploited circadian metabolism using both systems and synthetic biology tools, resulting in the obligatorily coupling of fumarate to either biomass or energy production. Resorting to laboratory evolution experiments, we show that this engineering approach is more stable than conventional methods.

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Metformin induces oxidative stress-mediated apoptosis without the blockade of glycolysis in h4iie hepatocellular carcinoma cells.

Park, D. (2019). Biological and Pharmaceutical Bulletin, 42(12), 2002-2008.

Metformin, a widely prescribed anti-diabetic drug, also exerts anti-cancer effects in different types of cancers. Although a number of molecular mechanisms have been suggested, the metabolic features underlying metformin’s anti-cancer activity is not fully understood enough. Because cancer cells have been known to prefer inefficient aerobic glycolysis to support their proliferation, it is important to clarify by which metformin affects metabolism to suppress the proliferation of cancer cells. Here, we report the metabolic changes induced by metformin and its relevance to the induction of apoptosis in H4II rat hepatocellular carcinoma cells. H4IIE cells were treated with metformin and other reagents in culture media with various nutritional compositions. Glutamine as well as pyruvate enhanced the viability of H4IIE cells in glucose-deprived conditions. Protective effects of glucose and pyruvate were comparable at same concentrations (5 mM). Metformin induced apoptosis irrespective of any nutritional conditions. Glucose consumption and lactate production were stimulated by metformin. Inhibition of glycolysis by 2-deoxyglucose suppressed the metformin-induced lactate production but additively enhanced metformin’s pro-apoptotic effect. These results indicate that metformin does not interfere but accelerate glycolysis. Unexpectedly, the production of reactive oxygen species (ROS) was markedly stimulated by metformin. A potent antioxidant, N-acetylcysteine (NAC) suppressed all pro-apoptotic changes as well as ROS generation induced by metformin. Taken together, metformin does not interfere with glycolysis but promotes apoptosis by enhancing oxidative stress.

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Infection/inflammation-associated preterm delivery within 14 days of presentation with symptoms of preterm labour: A multivariate predictive model.

Amabebe, E., Reynolds, S., He, X., Wood, R., Stern, V. & Anumba, D. O. (2019). PLoS One, 14(9), e0222455.

Multi-marker tests hold promise for identifying symptomatic women at risk of imminent preterm delivery (PTD, <37 week’s gestation). This study sought to determine the relationship of inflammatory mediators and metabolites in cervicovaginal fluid (CVF) with spontaneous PTD (sPTD) and delivery within 14 days of presentation with symptoms of preterm labour (PTL). CVF samples from 94 (preterm = 19, term = 75) singleton women with symptoms of PTL studied between 19+0-36+6 weeks’ gestation were analysed for cytokines/chemokines by multiplexed bead-based immunoassay, while metabolites were quantified by enzyme-based spectrophotometry in a subset of 61 women (preterm = 16, term = 45). Prevalence of targeted vaginal bacterial species was determined for 70 women (preterm = 14, term = 66) by PCR. Overall, 10 women delivered within 14 days of sampling. Predictive capacities of individual biomarkers and cytokine-metabolite combinations for sPTD and delivery within 14 days of sampling were analysed by logistic regression models and area under the receiver operating characteristic curve. Fusobacterium sp., Mubiluncus mulieris and Mycoplasma hominis were detected in more preterm-delivered than term women (P<0.0001), while, Mcurtisii was found in more term-delivered than preterm women (P<0.0001). RANTES (0.91, 0.65-1.0), IL-6 (0.79, 0.67-0.88), and Acetate/Glutamate ratio (0.74, 0.61-0.85) were associated with delivery within 14 days of sampling (AUC, 95% CI). There were significant correlations between cytokines and metabolites, and several cytokine-metabolite combinations were associated with sPTD or delivery within 14 days of sampling (e.g. L/D-lactate ratio+Acetate/Glutamate ratio+IL-6: 0.84, 0.67-0.94). Symptomatic women destined to deliver preterm and within 14 days of sampling express significantly higher pro-inflammatory mediators at mid to late gestation. In this cohort, IL-6, Acetate/Glutamate ratio and RANTES were associated with delivery within 14 days of sampling, consistent with their roles in modulating infection-inflammation-associated preterm labour in women presenting with symptoms of preterm birth. Replication of these observations in larger cohorts of women could show potential clinical utility.

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Respiratory and low-temperature sweetening responses of fresh-cut potato (Solanum tuberosum L.) tubers to low oxygen.

Ellis, G. D., Knowles, L. O. & Knowles, N. R. (2019). Postharvest Biology and Technology, 156, 110937.

Enzymatic browning (EB) has impeded the commercialization of fresh-cut (FC) potato. However, recently introduced Innate®-engineered cultivars with silenced polyphenol oxidase (PPO) have overcome this obstacle. As a supplement to refrigeration, low O2 atmosphere may extend the shelf-life of FC potato by reducing wound-induced respiration, associated dry matter loss, EB, and low-temperature sweetening (LTS). Determining the O2 concentration that minimizes these deteriorative physiological processes without invoking anaerobic metabolism is prerequisite to designing effective modified atmosphere packaging (MAP). Accordingly, FC tubers of cultivars Russet Burbank (RB), Ranger Russet (RR), and their Innate® counterparts, CultivATE®, GenerATE®, and GlaciATE®, were stored (4°C) in ten O2 atmospheres ranging from 0 to 21 kPa to determine the lower oxygen limit (LOL) for aerobic respiration and effects on LTS and EB. FC tissue stored at 21 kPa O2 displayed a prominent cold-induced respiratory acclimation response (RAR), characterized by an initial decline in respiration rate followed by a steady increase through 48 h. The RAR decreased with O2 and was extinguished at ≤1.5 kPa. Lowering O2 from 21-7 kPa had little effect on tissue respiration; however, rates fell from 4.23 to 3.41 μg kg−1 s−1 as O2 decreased from 7 to 3.5 kPa, followed by a further 79% reduction to 0.70 μg kg−1 s−1 at 0 kPa O2. Tissue lactate profiles revealed the onset of anaerobic metabolism at ca 1.5 kPa O2 for all cultivars. Importantly, lactate and ethanol accumulation were negligible through 16 d at ≥2 kPa O2 (= LOL) but increased considerably at ≤1 kPa. Low O2 attenuated the cold-induced synthesis of sucrose and reducing sugars in FC tissue from RB, and sucrose from GlaciATE® tubers in which acid invertase is silenced. Enzymatic browning of FC RB and RR tubers was inhibited by anoxia, but extensive at ≥0.5 kPa O2. FC Innate® tubers exhibited only minor EB at all O2 concentrations. Collectively, these data inform the further development of MAP for FC potato.

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Use of spent brewer's yeast in L‐(+) lactic acid fermentation.

Pejin, J., Radosavljević, M., Kocić‐Tanackov, S., Marković, R., Djukić‐Vuković, A. & Mojović, L. (2019). Journal of the Institute of Brewing, 125(3), 357-363.

The application of by‐products from the brewing industry in lactic acid (LA) production was investigated in order to replace expensive nitrogen sources (such as yeast extract) with cheaper and renewable nitrogenous materials such as brewer's yeast (BY). In this study, brewer's spent grain (BSG) hydrolysate was used for L‐(+)‐LA fermentation by Lactobacillus rhamnosus ATCC 7469. The effect of pH control during the fermentation and the addition of various BY contents (5-50 g/L) in BSG hydrolysate on fermentation parameters was evaluated. BY addition significantly increased free amino nitrogen (FAN) concentration (by 25.2% at 5 g/L to 616% at 50 g/L). A strong positive correlation between FAN concentration in the hydrolysate and concentration of L‐(+)‐LA produced was observed (correlation coefficient of 0.913). A high cell viability of L. rhamnosus ATCC 7469 (1.95–3.32 × 109 CFU/mL at the end of fermentation) was achieved in all fermentations with the addition of brewer's yeast. The addition of BY increased L‐(+)‐lactic acid yield and volumetric productivity up to 8.4% (5 g/L) and 48.3% (50 g/L). The highest L‐(+)‐LA yield (89%) and volumetric productivity (0.89 g/L h−1) were achieved in fermentation of BSG hydrolysate with 50 g/L of BY.

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Anoxia tolerance in the sea cucumbers Parastichopus californicus and Cucumaria miniata reflects habitat use.

Weinrauch, A. M. & Blewett, T. A. (2019). Journal of Experimental Marine Biology and Ecology, 520, 151203.

Natural fluctuations in dissolved oxygen are an important physiological challenge faced by marine organisms. This is particularly true of intertidal species that may become trapped in bodies of water where oxygen becomes rapidly depleted, or which may be emersed and exposed to elevated oxygen, but without respiratory organs adapted for uptake of oxygen from air. We sought to determine whether oxygen handling approaches differ in two species of sea cucumber, Cucumaria miniate and Parastichopus californicus, which inhabit distinct niches that vary in dissolved oxygen profiles. C. miniate occupies the rocky intertidal zone and thus experiences daily oscillations in oxygen content, whereas P. californicus inhabits the more oxygen stable, subtidal zone. Using closed chamber respirometry, our data showed that C. miniate have a higher basal metabolic rate than P. californicus, attributed to continuous filter-feeding, a phenomenon that contrasts with the sporadic feeding habits of the sediment-feeding P. californicus. Exposure to 6-h of anoxia or 6-h of emersion had no effect on coelomic fluid glucose concentrations in either species, however P. californicus body wall glucose concentrations increased ~6-fold relative to immersed normoxic controls under both treatments. Lactate production, a marker of anaerobic metabolism, was unaltered in both species under both anoxia and emersion conditions. These data demonstrate that habitat influences anoxia tolerance in sea cucumbers, with the intertidal C. miniate displaying a higher tolerance to both low dissolved oxygen and emersion, than the subtidal species, P. californicus.

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Effect of mixed species alcoholic fermentation on growth and malolactic activity of lactic acid bacteria.

Englezos, V., Cachón, D. C., Rantsiou, K., Blanco, P., Petrozziello, M., Pollon, M., Giacosa, S., Segade, S. R., Rolle, L. & Cocolin, L. (2019). Applied Microbiology and Biotechnology, 103(18), 7687-7702.

In recent years, there is an increasing interest from the winemaking industry for the use of mixed fermentations with Starmerella bacillaris (synonym Candida zemplinina) and Saccharomyces cerevisiae, due to their ability to modulate metabolites of oenological interest. The current study was carried out to elucidate the effect of this fermentation protocol on the growth and malolactic activity of lactic acid bacteria (LAB) used for malolactic fermentation (MLF) and on the chemical and volatile profile of Nebbiolo wines and their chromatic characteristics. To this end, two LAB species, namely Lactobacillus plantarum and Oenococcus oeni, were inoculated at the beginning and at the end of the alcoholic fermentation (AF) performed by pure and mixed yeast using the abovementioned yeasts. The different yeast inoculation protocols and the combination of species tested influenced greatly the interactions and behavior of the inoculated yeasts and LAB during AF and MLF. For both LAB species, inoculation timing was critical to how rapidly MLF started and finished. Fermentation inoculated with L. plantarum, at the beginning of the AF, completed MLF faster than those inoculated with O. oeni. The presence of Starm. Bacillaris in mixed fermentation promoted LAB growth and activity, in particular, O. oeni. Furthermore, LAB species choice had a greater impact on the volatile and chromatic profile of the wines than inoculation time. These findings reveal new knowledge about the importance of LAB species choice and inoculation time to ensure fast MLF completion and to improve wine characteristics in mixed fermentation with Starm. Bacillaris and S. cerevisiae.

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Assessment of β-glucans, phenols, flavor and volatile profiles of hulless barley wine originating from highland areas of China.

Zhang, K., Yang, J., Qiao, Z., Cao, X., Luo, Q., Zhao, J., Wang, F. & Zhang, W. (2019). Food Chemistry, 293, 32-40.

Low alcohol hulless barley wine (HW) is a popular beverage among the highland areas in China. It is known to have several health benefits due to the presence of β-glucan and antioxidant compounds. Therefore, the total β-glucan content, total phenols and flavonoids of HW samples from the highland areas of Sichuan province and Tibet were determined in this study. The results indicated that HW is abundant in both β-glucan (54-76 mg/L) and phenolic compounds (131-178 mg/L). Moreover, this study also investigated the flavor and aroma characteristics of HW samples. A total of forty six volatile aroma substances were identified by GC-MS. The HWs could be classified into three distinct groups in terms of the region of origin according to the results of PCA based on the GC-MS data. These findings provide a useful foundation for further study of the health benefits and the flavor characteristics of HW in highland areas.

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Acetate metabolism and the inhibition of bacterial growth by acetate.

Pinhal, S., Ropers, D., Geiselmann, J. & de Jong, H. (2019). Journal of Bacteriology, 201(13).

During aerobic growth on glucose, Escherichia coli excretes acetate, a mechanism called “overflow metabolism.” At high concentrations, the secreted acetate inhibits growth. Several mechanisms have been proposed for explaining this phenomenon, but a thorough analysis is hampered by the diversity of experimental conditions and strains used in these studies. Here, we describe the construction of a set of isogenic strains that remove different parts of the metabolic network involved in acetate metabolism. Analysis of these strains reveals that (i) high concentrations of acetate in the medium inhibit growth without significantly perturbing central metabolism; (ii) growth inhibition persists even when acetate assimilation is completely blocked; and (iii) regulatory interactions mediated by acetyl-phosphate play a small but significant role in growth inhibition by acetate. The major contribution to growth inhibition by acetate may originate in systemic effects like the uncoupling effect of organic acids or the perturbation of the anion composition of the cell, as previously proposed. Our data suggest, however, that under the conditions considered here, the uncoupling effect plays only a limited role.

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Role of adiponectin in the metabolism of skeletal muscles in collagen VI-related myopathies.

Gamberi, T., Magherini, F., Mannelli, M., Chrisam, M., Cescon, M., Castagnaro, S., Modesti, A., Braghetta, P. & Fiaschi, T. (2019). Journal of Molecular Medicine, 97(6), 793-801.

The role of adiponectin has been particularly deepened in diabetic muscles while the study of adiponectin in hereditary myopathies has been marginally investigated. Here, we report the study about adiponectin effects in Col6a1−/− (collagen VI–null) mice. Col6a1−/− mice show myophatic phenotype closer to that of patients with Bethlem myopathy, thus representing an excellent animal model for the study of this hereditary disease. Our findings demonstrate that Col6a1−/− mice have decreased plasma adiponectin content and diseased myoblasts have an impaired autocrine secretion of the hormone. Moreover, Col6a1−/− myoblasts show decreased glucose uptake and mitochondria with depolarized membrane potential and impaired functionality, as supported by decreased oxygen consumption. Exogenous addition of globular adiponectin modifies the features of Col6a1−/− myoblasts, becoming closer to that of the healthy myoblasts. Indeed, globular adiponectin enhances glucose uptake in Col6a1−/− myoblasts, modifies mitochondrial membrane potential, and restores oxygen consumption, turning closer to those of wild-type myoblasts. Finally, increase of plasma adiponectin level in Col6a1−/− mice is induced by fasting, a condition that has been previously shown to lead to the amelioration of the dystrophic phenotype. Collectively, our results demonstrate that exogenous replenishment of adiponectin reverses metabolic abnormalities observed in Col6a1−/− myoblasts.

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Utilization of brewing and malting by-products as carrier and raw materials in l-(+)-lactic acid production and feed application.

Radosavljević, M., Pejin, J., Pribić, M., Kocić-Tanackov, S., Romanić, R., Mladenović, D., Djukić-Vuković, A. & Mojović, L. (2019). Applied Microbiology and Biotechnology, 103(7), 3001-3013.

Application of agro-industrial by-products for the production of lactic acid was studied in this paper. Brewer’s spent grain (BSG), malt rootlets (MR), brewer’s yeast (BY), and soy lecithin (SL) were used as raw materials in L-(+)-LA fermentation by free and immobilized Lactobacillus rhamnosus ATCC 7469. The BSG, solid remains after BSG and MR hydrolysis (BSGMRSR), and MR were evaluated as carriers for batch and repeated batch fermentations with immobilized cells. During batch fermentations with immobilized cells, high cell viability (10 to 11 log CFU/g) was achieved on all carriers. In batch fermentation with BSG as a carrier, the highest LA yield of 93.79% and volumetric productivity of 1.15 g/L/h were obtained. Furthermore, very high LA yield (95.46%), volumetric productivity (1.98 g/L/h) and L. rhamnosus viability (11.5 log CFU/g) were achieved in repeated batch fermentations with the cells immobilized on this carrier. The immobilized cells showed high survival rate (94-95%) during exposure to simulated gut condition. Based on the analysis of BSGMRSR, and BY solid remains, and on in vitro evaluation of the probiotic characteristics of immobilized cells, it was observed that they could satisfy the recommendations for high-quality feed preparation.

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Potential properties of Lactobacillus plantarum F-10 as a bio-control strategy for wound infections.

Onbas, T., Osmanagaoglu, O. & Kiran, F. (2019). Probiotics and Antimicrobial Proteins, 11(4), 1110-1123.

In this study, Lactobacillus plantarum  F-10, a promising probiotic strain isolated from fecal microbiota of healthy breastfed infant, was assessed as a bio-control strategy for wound infections. Pseudomonas aeruginosa = PAO1/ATCC 27853, methicillin-resistant Staphylococcus aureus ATCC 43300, and their hospital-derived strains isolated from skin chronic wound samples were used as important skin pathogens. The cell-free extract (CFE) of the strain F-10 was shown to inhibit the growth of all pathogens tested, while no inhibition was observed when CFE was neutralized. The strain displayed no hemolysis and exhibited a strong auto-aggregating phenotype (51.48 ± 1.45%, 5 h) as well as co-aggregation. Antibiotic resistance profile was found to be safe according to the European Food Safety Authority. Biofilm formation was measured by crystal violet assay and visualized with scanning electron microscopy and confocal laser scanning microscopy. One hundred percent reduction in biofilm formation of all pathogens tested was obtained by sub-MIC value (12.5 mg/ml) of CFE following 24-h co-incubation. Inhibition of quorum sensing–controlled virulence factors (motility, protease and elastase activity, production of pyocyanin and rhamnolipid) of P. aeruginosa strains was also observed. DPPH radical scavenging activity of the CFE was determined as 88.57 ± 0.49%. In conclusion, our results suggest that L. plantarum F-10 may represent an alternative bio-control strategy against skin infections with its antimicrobial, anti-biofilm, anti-quorum sensing, and antioxidant activity.

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