Physiological and molecular changes of onion (Allium cepa L.) seeds under different aging conditions.

BACKGROUND: Onion seeds have limited storage capacity compared to other vegetable seeds. It is crucial to identify the mechanisms that induce tolerance to storage conditions and reduce seed deterioration. To address this goal, an experiment was conducted to evaluate changes in germination, biochemical, physiological, and molecular characteristics of onion seed landraces (Horand, Kazerun landraces and Zargan cultivar) at different aging levels (control, three-days and six-days accelerated aging, and natural aging for one year). RESULTS: The findings suggest that there was an increase in glucose, fructose, total sugar, and electrolyte leakage in the Horand (HOR), Kazerun (KAZ) landraces, and Zarghan (ZAR) cultivar, with Kazerun exhibiting the greatest increase. The percentage and rate of germination of Kazerun decreased by 54% and 33%, respectively, in six-day accelerated aging compared to the control, while it decreased by 12% and 14%, respectively, in Horand. Protein content decreased with increasing levels of aging, with a decrease of 26% in Kazerun landrace at six days of aging, while it was 16% in Horand landrace. The antioxidant activities of catalase, superoxide dismutase, and glutathione peroxidase decreased more intensively in Kazerun. The expression of AMY1, BMY1, CTR1, and NPR1 genes were lower in Kazerun landraces than in Horand and Zargan at different aging levels. CONCLUSIONS: The AMY1, BMY1, CTR1, and NPR1 genes play a pivotal role in onion seed germination, and their downregulation under stressful conditions has been shown to decrease germination rates. In addition, the activity of CAT, SOD, and GPx enzymes decreased by seed aging, and the amount of glucose, fructose, total sugar and electrolyte leakage increased, which ultimately led to seed deterioration. Based on the results of this experiment, it is recommended to conduct further studies into the molecular aspects involved in onion seed deterioration. More research on the genes related to this process is suggested, as well as investigating the impact of different priming treatments on the genes expression involved in the onion seed aging process.

Effective drying processes for Taikor (Garcinia pedunculata Roxb.) fruit by ultrasound-assisted osmotic pretreatment: Analysis of quality and kinetic models.

The present study aimed to analyze and establish an effective combination of ultrasound and immersion pretreatment processes for drying Taikor (Garcinia pendunculata Roxb.) fruits. Taikorslices were first immersed in 10 % sucrose, fructose, and glucose solution. Then, the immersed slices were treated in an ultrasonic bath at 30 °C for 10, 20, and 30 min. Drying operations were carried out at 50, 60, and 70 °C, with a fixed relative humidity of 30 %. The Page, Newton, Henderson and Pabis, and Weibull distribution models were fitted to the obtained drying data to determine the best kinetic model that effectively describes the drying properties ofTaikor. After drying operations, changes in quality parameters, e.g., ß-carotene, vitamin C, B vitamins, color, antioxidant activities, and microbial loads, were measured to obtain the best drying temperature and the most effective pretreatment combination with minimum loss of nutrients of the sample. Among different kinetic models, both Page and Weibull distribution models showed the best R2 values of 0.9867 and 0.9366, respectively. The chemical properties were preserved to the greatest extent possible by drying at 50 °C with glucose pretreatment. The color parameters were better preserved by fructose pretreatment. Sonication time also had profound effect on the quality parameters of dried Taikor slices. However, higher temperature drying required a shorter time for drying and exhibited better performance in microbial load reduction. This study's findings will help to establish an effective drying condition forGarcinia pedunculatafruits.

Dry heat whole Sorghum BRS 305 flour modulate satiety and improves antioxidant response in brain of Wistar rats fed with high-fat high-fructose diet.

Sorghum BRS 305 (Sorghum bicolor L. Moench) is a cereal with high tannins and anthocyanins content and keep better the resistant starch when submitted to dry heat treatment. Our objective was to investigate the effects of BRS 305 dry heat treatment whole sorghum flour on satiety and antioxidant response in brain and adipose tissue of Wistar rats fed with a high fat high fructose diet (HFHF). Male Wistar rats were divided in two groups: control (n = 8) and HFHF (n = 16) for eight weeks. After, animals of HFHF group were divided: HFHF (n = 8) and HFHF + BRS 305 sorghum whole flour (n = 8), for 10 weeks. Sorghum consumption reduced gene expression of leptin, resistin, and endocannabinoid receptor 1 type (CB1) in adipose and brain tissues compared to HFHF group. In brain, sorghum consumption also promotes reduction in neuropeptide Y (NPY) gene expression. BRS305 sorghum consumption improved gene expression of sirtuin-1 (SIRT1) in adipose tissue, and in the brain increased heat shock protein 72 (HSP72), erythroid-derived nuclear factor 2 (NRF2), peroxisome proliferator-activated receptor alpha (PPARα), superoxide dismutase (SOD) and catalase activity compared to HFHF. In silicoanalysis showed interaction with PPARα, CB1, and leptin receptors. Advanced glycation end products (AGEs) concentrations in group HFHF + sorghum did not differ from HFHF group. Advanced glycation end products receptors (RAGEs) concentrations did not differ among experimental groups. Then, BRS 305 sorghum submitted to dry treatment was able to modulate gene expression of markers related to satiety and improve antioxidant capacity of rats fed with HFHF diet.

Validation of an Eco-Friendly Automated Method for the Determination of Glucose and Fructose in Wines.

Fermentable sugar dosage helps oenologists to establish a harvest's moment and control the fermentation process of the musts. The official analyses recommended for their determination are long, laborious, and must be carried out by specialized personnel. On the contrary, instrumental analysis automation limits human errors, increases precision, and reduces the time and cost of the analyses. In the food production sector, to use methods other than those recommended by supranational bodies in official reports, it is necessary to validate the analytical processes to establish the conformity of the results between the new methods and the reference ones. This work validated an automated enzymatic apparatus to determine the sum of glucose and fructose levels in wine samples. The validation was carried out on wine samples (dry red wine, dry white wine, moderately sweet wine, and sweet wine) containing different sugar concentrations by comparing data obtained using the OIV-MA-AS311-02 method performed by a specialized operator (reference method) and the same method performed by an automated apparatus. The difference between the results' means obtained with the two procedures was significant. Nevertheless, the automated procedure was considered suitable for the intended use since the differences between the averages were lower than the measurement uncertainty at the same concentration, and the repeatability results were better for the automated procedure than the reference method.

Determination of quality traits and possible adulteration of molasses using FT-IR spectroscopy: A study from Turkish market.

We aimed to develop portable Fourier transform infrared (FT-IR) spectroscopy-based prediction algorithms for the key quality characteristics (soluble solids, water activity, pH, sucrose, glucose, fructose, fructose/glucose, hydroxymethylfurfural) of various types of molasses, establish their legitimacy, and create a model to separate them based on their botanical origin. Samples labeled as carob (n = 27), grape (n = 24), Juniper (n = 13), and mulberry (n = 12) were purchased from different local markets in Turkey. Labeling issues were revealed in five carob and seven grape molasses, and those samples classified as non-authentic by the FT-IR algorithms were corroborated by reference analysis. Partial least squares regression models generated to predict the key quality traits of Turkish molasses demonstrated excellent correlation with reference analysis (R2Val ≥ 0.96) and low standard error of prediction (SEP ≤ 2.88). The FT-IR sensor provided a feasible approach for molasses testing to assess its quality through manufacturing and storage, also provided a powerful tool to -ensure proper product labeling.

Determination of Sugars and Acids in Grape Must Using Miniaturized Near-Infrared Spectroscopy.

An automatic determination of grape must ingredients during the harvesting process would support cellar logistics and enables an early termination of the harvest if quality parameters are not met. One of the most important quality-determining characteristics of grape must is its sugar and acid content. Among others, the sugars in particular determine the quality of the must and wine. Chiefly in wine cooperatives, in which a third of all German winegrowers are organized, these quality characteristics serve as the basis for payment. They are acquired upon delivery at the cellar of the cooperative or the winery and result in the acceptance or rejection of grapes and must. The whole process is very time-consuming and expensive, and sometimes grapes that do not meet the quality requirements for sweetness, acidity, or healthiness are destroyed or not used at all, which leads to economic loss. Near-infrared spectroscopy is now a widely used technique to detect a wide variety of ingredients in biological samples. In this study, a miniaturized semi-automated prototype apparatus with a near-infrared sensor and a flow cell was used to acquire spectra (1100 nm to 1350 nm) of grape must at defined temperatures. Data of must samples from four different red and white Vitis vinifera (L.) varieties were recorded throughout the whole growing season of 2021 in Rhineland Palatinate, Germany. Each sample consisted of 100 randomly sampled berries from the entire vineyard. The contents of the main sugars (glucose and fructose) and acids (malic acid and tartaric acid) were determined with high-performance liquid chromatography. Chemometric methods, using partial least-square regression and leave-one-out cross-validation, provided good estimates of both sugars (RMSEP = 6.06 g/L, R2 = 89.26%), as well as malic acid (RMSEP = 1.22 g/L, R2 = 91.10%). The coefficient of determination (R2) was comparable for glucose and fructose with 89.45% compared to 89.08%, respectively. Although tartaric acid was predictable for only two of the four varieties using near-infrared spectroscopy, calibration and validation for malic acid were accurate for all varieties in an equal extent like the sugars. These high prediction accuracies for the main quality determining grape must ingredients using this miniaturized prototype apparatus might enable an installation on a grape harvester in the future.

Determination of sugar content in honey using LC-Raman and programmable pump-Raman methods.

We combined (i) liquid chromatography and Raman spectrometry (LC-Raman) and (ii) programmable pump and Raman spectrometry (PP-Raman) to separate and identify compounds in a mixture. These techniques were applied to conduct a quantitative analysis of the sugars in honey. The spectral and temporal axes of the LC-Raman data were analyzed using the MCR-ALS analysis procedure, which enabled the separation and identification of four sugars (glucose, fructose, sucrose, and trehalose). The PP-Raman method was employed to examine the sugar concentration dependence of the intensity pattern of the Raman spectrum, and the linear concentration dependence of the intensity was obtained. The sugar contents were quantitatively determined from the integrated area of the elution peaks. The result was consistent with those derived from mass spectrometry and previous studies. The origin of the errors in the derived sugar contents is discussed. Our study presents a novel quantitative LC-Raman spectrometric method that does not rely on resonance or surface enhancement effects.

Authentication and Geographical Characterisation of Italian Grape Musts through Glucose and Fructose Carbon Isotopic Ratios Determined by LC-IRMS.

The authenticity of grape musts is normally checked through a time-consuming stable isotopic analysis of carbon (δ13C) after fermentation and distillation by following the official OIV MA AS-312-06 method. In this study, the alternative use of a technique based on δ13C isotopic analysis of the major sugars of the grape must by liquid chromatography coupled with isotope ratio mass spectrometry (LC-IRMS) is provided. It allows not only the detection of the fraudulent addition to grape must of exogenous glucose and fructose deriving from C4 plants but also the characterisation of it based on its geographical origin. In order to discriminate between musts from different areas of Italy, a preliminary dataset was considered; the δ13C isotopic ratios of glucose and fructose of around 100 authentic samples were analysed. The two analysed parameters, ranging from -29.8‱ to -21.9‱, are well correlated (R2 = 0.7802) and the northern regions showed significantly more negative δ13C values for both sugars than the rest of the dataset.

Arginine-fructose-glucose from red ginseng extract reduces stiffness of keratin fiber in corneocyte of skin.

PURPOSE: The moisture content of the stratum corneum of the skin changes depending on the external environment. The structure of keratinous fiber protein in corneocyte of the skin changes depending on the amount of moisture. As the moisture decreases, the population of the alpha-helix increases, the beta-sheet deceases, and the stiffness increases accordingly. Here, we investigated the effect of humectants from ginseng on the keratin structure. METHODS: Corneocyte was prepared from dry porcine skin with disc tape and measured through ATR-FT-IR. The signal from amide I of the keratin protein in corneocyte was detected, and the change in the ratio of alpha-helix and beta-sheet was calculated. The test samples were treated on the exfoliated corneocyte, and the degree of change was checked. RESULT: Arginine-fructose-glucose (AFG)-enriched extract of red ginseng was effective in changing the keratin structure and was superior to humectants such as glycerin. However, arginine, mono sugar were not effective, and the AFG form in which two sugars were bound to one amino acid could perform its function. CONCLUSION: The present study suggests that AFG, when applied to cosmetics, is expected to improve skin texture in a different way from existing moisturizers represented by glycerin by reducing the alpha-helix structure of corneocyte keratin.

Sugar Concentration, Nitrogen Availability, and Phylogenetic Factors Determine the Ability of Acinetobacter spp. and Rosenbergiella spp. to Grow in Floral Nectar.

The floral nectar of angiosperms harbors a variety of microorganisms that depend predominantly on animal visitors for their dispersal. Although some members of the genus Acinetobacter and all currently known species of Rosenbergiella are thought to be adapted to thrive in nectar, there is limited information about the response of these bacteria to variation in the chemical characteristics of floral nectar. We investigated the growth performance of a diverse collection of Acinetobacter (n = 43) and Rosenbergiella (n = 45) isolates obtained from floral nectar and the digestive tract of flower-visiting bees in a set of 12 artificial nectars differing in sugar content (15% w/v or 50% w/v), nitrogen content (3.48/1.67 ppm or 348/167 ppm of total nitrogen/amino nitrogen), and sugar composition (only sucrose, 1/3 sucrose + 1/3 glucose + 1/3 fructose, or 1/2 glucose + 1/2 fructose). Growth was only observed in four of the 12 artificial nectars. Those containing elevated sugar concentration (50% w/v) and low nitrogen content (3.48/1.67 ppm) were limiting for bacterial growth. Furthermore, phylogenetic analyses revealed that the ability of the bacteria to grow in different types of nectar is highly conserved between closely related isolates and genotypes, but this conservatism rapidly vanishes deeper in phylogeny. Overall, these results demonstrate that the ability of Acinetobacter spp. and Rosenbergiella spp. to grow in floral nectar largely depends on nectar chemistry and bacterial phylogeny.

Formation and reduction of furan in pumpkin puree by precursors, antioxidants, sterilization and reheating.

The formation and mitigation of furan in pumpkin puree (PP) were studied during the complete process of producing PP. The content of furan was determined using headspace solid-phase microextraction combined by GC-MS analysis. Various PP samples added with precursors (glucose, fructose, linoleic acid, linolenic acid, ß-carotene, ascorbic acid, glutamic acid, alanine, and serine) showed increased furan formation (30.81 âˆ¼ 94.45 µg/kg) compared with the control (30.81 µg/kg), with ß-carotene resulting in the formation of the largest amount of furan. The effects of antioxidants, such as caffeic acid, chlorogenic acid, quercetin, and butylated hydroxytoluene, on the reduction of furan in PP containing ß-carotene were also investigated. All antioxidants showed significant reduction of furan. During sterilizing, the content of furan was considerably affected by temperature but not heating time. Reheating PP samples using a microwave oven, water bath, or open pot, revealed that open-pot reheating was the most effective for reducing furan (10.28 âˆ¼ 11.72 µg/kg).

Non-targeted detection of grape molasses adulteration with sugar and apple molasses by mid-infrared spectroscopy coupled to independent components analysis.

In the light of the current food security crisis, food adulteration has resurfaced on the international scene, inflicting potential safety issues and leading more and more consumers into deception. This situation led food control actors to remobilise their potential to face this problem, particularly in terms of analytical chemistry competencies. Similar to honey, grape molasses may be considered very likely to be adulterated leading to quality and authenticity issues, especially in the Eastern Mediterranean, where it is widely consumed as a traditional sweetener. This work reports the use of attenuated total reflectance-mid-infrared spectroscopy (ATR-MIR) coupled to chemometrics, as an alternative to complex, expensive and time-consuming analytical techniques, in the aim of detecting fraudulent glucose, fructose, sucrose and apple molasses additions to pure grape molasses. After collecting a widespread unadulterated grape molasses database, spiked samples with increasing concentrations (w/w) of the selected adulterants were prepared. In order to establish a qualitative model, whose potential is to detect adulteration and discriminate between the different adulterants, samples underwent ATR-MIR analyses without any prior preparation, and the collected spectral data were subjected to independent components analysis (ICA), where Random_ICA was used to retrieve the optimal number of independent components (ICs). Thereupon, the extraction of seven ICs allowed the establishment of a qualitative model with a clear discrimination between molasses adulterated with fructose, sucrose and glucose syrup, relying on MIR specific signals and incorporated ratios of the different adulterants. However, it failed in detecting apple molasses adulteration, calling for the development of a different analytical approach. The developed model underwent a verification step using a control set recorded on a different spectrometer, proving its potential to provide reproducible discrimination and classification rates.

Selective detection of starch in banana fruit with Raman spectroscopy.

To develop an analytical method for rapid quantification of starch in agricultural produce, we measured the Raman spectra of ripening banana fruit and compared the obtained data to those of standard starch, sugar, and fiber chemical samples. Standard starches exhibited distinctive Raman bands, which were similar to the spectral features in green banana before ripening. Moreover, these banana-derived Raman bands gradually weakened during 10 d of storage. Standard sugars (glucose, sucrose, and fructose) exhibited Raman bands in both solid and liquid states, whereas standard fibers exhibited broad spectra and no such bands. Although the sugar content increased, no sugar bands were observed in the banana fruit even after ripening. A correlation was found between the Raman bands and starch content obtained by chemical analysis. These results demonstrate that Raman spectroscopy can selectively provide information regarding starch in banana fruit and be applied as an analytical method for rapid starch quantification.

Monitoring glycolytic dynamics in single cells using a fluorescent biosensor for fructose 1,6-bisphosphate.

Cellular metabolism is regulated over space and time to ensure that energy production is efficiently matched with consumption. Fluorescent biosensors are useful tools for studying metabolism as they enable real-time detection of metabolite abundance with single-cell resolution. For monitoring glycolysis, the intermediate fructose 1,6-bisphosphate (FBP) is a particularly informative signal as its concentration is strongly correlated with flux through the whole pathway. Using GFP insertion into the ligand-binding domain of the Bacillus subtilis transcriptional regulator CggR, we developed a fluorescent biosensor for FBP termed HYlight. We demonstrate that HYlight can reliably report the real-time dynamics of glycolysis in living cells and tissues, driven by various metabolic or pharmacological perturbations, alone or in combination with other physiologically relevant signals. Using this sensor, we uncovered previously unknown aspects of ß-cell glycolytic heterogeneity and dynamics.

Differentiation and Quantitation of Coeluting Isomeric Amadori and Heyns Peptides Using Sugar-Specific Fragment Ion Ratios.

d-glucose and d-fructose present in blood, tissues, and organs of all mammals can react with amino groups, leading to glucated (Amadori) and fructated (Heyns) products, i.e., proteins glycated at lysine residues. While typically present at low concentration in humans, metabolic diseases including diabetes elevate sugar levels, favoring glycation and consecutive reactions leading to advanced glycation end products (AGEs) linked to diabetic complications and cardiovascular diseases. Analytical methods able to differentiate and to individually quantify Amadori- and Heyns-modified proteins in complex sample mixtures, e.g., serum, are still very limited. Here, we show that the reported and supposedly specific neutral losses displayed in tandem mass spectra of Heyns peptides cannot be used for a reliable differentiation as they were also observed for Amadori peptides. However, the combination of several neutral loss signals in fragment ion ratios at both precursor and fragment ion signals allowed the differentiation and relative quantitation of coeluting isomeric Amadori and Heyns peptides at different concentrations and peptide ratios. This was also true for digested human plasma. Thus, the presented strategy allows the quantitation of Amadori and Heyns peptides in complex samples, especially by spiking isotope-labeled peptides. This will allow searching for glucated and fructated biomarkers in clinical samples.

Fructose and methylglyoxal-induced glycation alters structural and functional properties of salivary proteins, albumin and lysozyme.

Glycation process refers to reactions between reduction sugars and amino acids that can lead to formation of advanced glycation end products (AGEs) which are related to changes in chemical and functional properties of biological structures that accumulate during aging and diseases. The aim of this study was to perform and analyze in vitro glycation by fructose and methylglyoxal (MGO) using salivary fluid, albumin, lysozyme, and salivary α-amylase (sAA). Glycation effect was analyzed by biochemical and spectroscopic methods. The results were obtained by fluorescence analysis, infrared spectroscopy (total attenuated reflection-Fourier transform, ATR-FTIR) followed by multivariate analysis of principal components (PCA), protein profile, immunodetection, enzymatic activity and oxidative damage to proteins. Fluorescence increased in all glycated samples, except in saliva with fructose. The ATR-FTIR spectra and PCA analysis showed structural changes related to the vibrational mode of glycation of albumin, lysozyme, and salivary proteins. Glycation increased the relative molecular mass (Mr) in protein profile of albumin and lysozyme. Saliva showed a decrease in band intensity when glycated. The analysis of sAA immunoblotting indicated a relative reduction in intensity of its correspondent Mr after sAA glycation; and a decrease in its enzymatic activity was observed. Carbonylation levels increased in all glycated samples, except for saliva with fructose. Thiol content decreased only for glycated lysozyme and saliva with MGO. Therefore, glycation of salivary fluid and sAA may have the potential to identify products derived by glycation process. This opens perspectives for further studies on the use of saliva, an easy and non-invasive collection fluid, to monitor glycated proteins in the aging process and evolution of diseases.

Comparative study of sugar extraction procedures for HPLC analysis and proposal of an ethanolic extraction method for plant-based high-protein ingredients.

BACKGROUND: The increasing importance of plant-based proteins in the food sector makes a reliable compositional analysis of plant-based high-protein ingredients a necessity. Specifically, the quantification of short-chain carbohydrates is relevant for multiple areas, including food product development, food labelling and fundamental food chemistry and food technology research. Commonly used extraction procedures for subsequent high-performance liquid chromatographic separation and quantification of short-chain carbohydrates have been discussed controversially regarding a range of complications that can potentially lead to inaccurate sugar determination. The present study compares the sugar levels in wheat flour and wholemeal wheat flour determined with different aqueous and ethanolic extraction procedures. These procedures included measures to prevent enzyme activity and microbial growth, which represent two of the most relevant challenges in sugar extraction from food samples. RESULTS: Differences in sugar levels (sum of sucrose/maltose, glucose and fructose) as high as 1.8% dry matter (wheat flour) were observed between the employed extraction procedures. Ethanolic extraction (80% ethanol in ultrapure water) with the use of the antimicrobial agent sodium azide but without Carrez clarification was identified as most promising for sugar determination in plant-based high-protein ingredients. CONCLUSION: A screening of high-protein ingredients derived from cereals (wheat gluten), pseudocereals (quinoa, amaranth, buckwheat) and legumes (soy, pea, lupin, lentil, carob, chickpea, faba bean) concerning their levels of sucrose, maltose, glucose and fructose confirmed the applicability of the chosen extraction procedure. © 2021 Society of Chemical Industry.

Inulin-grown Faecalibacterium prausnitzii cross-feeds fructose to the human intestinal epithelium.

Many chronic diseases are associated with decreased abundance of the gut commensal Faecalibacterium prausnitzii. This strict anaerobe can grow on dietary fibers, e.g., prebiotics, and produce high levels of butyrate, often associated to epithelial metabolism and health. However, little is known about other F. prausnitzii metabolites that may affect the colonic epithelium. Here, we analyzed prebiotic cross-feeding between F. prausnitzii and intestinal epithelial (Caco-2) cells in a "Human-oxygen Bacteria-anaerobic" coculture system. Inulin-grown F. prausnitzii enhanced Caco-2 viability and suppressed inflammation- and oxidative stress-marker expression. Inulin-grown F. prausnitzii produced excess butyrate and fructose, but only fructose efficiently promoted Caco-2 growth. Finally, fecal microbial taxonomy analysis (16S sequencing) from healthy volunteers (n = 255) showed the strongest positive correlation for F. prausnitzii abundance and stool fructose levels. We show that fructose, produced and accumulated in a fiber-rich colonic environment, supports colonic epithelium growth, while butyrate does not.

Validity of FFQ Estimates of Total Sugars, Added Sugars, Sucrose and Fructose Compared to Repeated 24-h Recalls in Adventist Health Study-2 Participants.

Sugar intake is a potentially important aspect of diet which has not previously been validated in the Adventist Health Study-2 (AHS-2). We sought to validate the food frequency questionnaire (FFQ) measurement of total sugars, added sugars, sucrose, and fructose against multiple 24-h dietary recalls (recalls) in AHS-2 participants. Food consumption data from a self-administered FFQ and six recalls from 904 participants were combined with nutrient profile data to estimate daily sugar intake. Validity was evaluated among all participants and by race. FFQ and recall means were compared and correlation coefficients (Spearman's, energy-adjusted log-transformed Pearson's, deattenuated Pearson's) were calculated. Mean total energy, total sugars, and fructose intake were higher in the FFQ, whereas added sugars and sucrose were higher in recalls. The energy-adjusted (log-transformed) deattenuated correlations among all participants were: total sugars (r = 0.42, 95% CI 0.32-0.52), added sugars (r = 0.50, 95% CI 0.36-0.59), sucrose (r = 0.32, 95% CI 0.23-0.42), and fructose (r = 0.50, 95% CI 0.40-0.59). We observed moderate validity for added sugars and fructose and low-moderate validity for total sugars and sucrose measured by the AHS-2 FFQ in this population. Dietary sugar estimates from this FFQ may be useful in assessing possible associations of sugars intake with health outcomes.

Recombinant protein production provoked accumulation of ATP, fructose-1,6-bisphosphate and pyruvate in E. coli K12 strain TG1.

BACKGROUND: Recently it was shown that production of recombinant proteins in E. coli BL21(DE3) using pET based expression vectors leads to metabolic stress comparable to a carbon overfeeding response. Opposite to original expectations generation of energy as well as catabolic provision of precursor metabolites were excluded as limiting factors for growth and protein production. On the contrary, accumulation of ATP and precursor metabolites revealed their ample formation but insufficient withdrawal as a result of protein production mediated constraints in anabolic pathways. Thus, not limitation but excess of energy and precursor metabolites were identified as being connected to the protein production associated metabolic burden. RESULTS: Here we show that the protein production associated accumulation of energy and catabolic precursor metabolites is not unique to E. coli BL21(DE3) but also occurs in E. coli K12. Most notably, it was demonstrated that the IPTG-induced production of hFGF-2 using a tac-promoter based expression vector in the E. coli K12 strain TG1 was leading to persistent accumulation of key regulatory molecules such as ATP, fructose-1,6-bisphosphate and pyruvate. CONCLUSIONS: Excessive energy generation, respectively, accumulation of ATP during recombinant protein production is not unique to the BL21(DE3)/T7 promoter based expression system but also observed in the E. coli K12 strain TG1 using another promoter/vector combination. These findings confirm that energy is not a limiting factor for recombinant protein production. Moreover, the data also show that an accelerated glycolytic pathway flux aggravates the protein production associated "metabolic burden". Under conditions of compromised anabolic capacities cells are not able to reorganize their metabolic enzyme repertoire as required for reduced carbon processing.