Production of the prolyl endoprotease (PEP) from Aspergillus sp. FSDE 16 by solid-state fermentation (SSF) and use for producing a gluten-free beer.

Beer is a beverage that contains gluten and cannot be consumed by people with celiac disease. In this context, the enzyme prolyl endoprotease (PEP) can be used to reduce the gluten content in beer. The present study aimed to produce the PEP from Aspergillus sp. FSDE 16 using solid-state fermentation with 5 conditions and comparing with a similar commercial enzyme produced from Aspergillus niger in the production of a gluten-free beer. The results of the performed cultures showed that during the culture, the most increased protease activity (54.46 U/mL) occurred on the 4th day. In contrast, for PEP, the highest activity (0.0356 U/mL) was obtained on the 3rd day of culture in condition. Regarding beer production, cell growth, pH, and total soluble solids showed similar behavior over the 7 days for beers produced without enzyme addition or with the addition of commercial enzyme and with the addition of the enzyme extract produced. The addition of the enzyme and the enzyme extract did not promote changes, and all the beers produced showed similar and satisfactory results, with acid pH between 4 and 5, total soluble solids ranging from 4.80 to 5.05, alcohol content ranging from 2.83% to 3.08%, and all beers having a dark character with deep amber and light copper color. Gluten removal was effectively using the commercial enzyme and the enzyme produced according to condition (v) reaching gluten concentrations equal to 17 ± 5.31 and 21.19 ± 11.28 ppm, respectively. In this way, the production of the enzyme by SSF and its application in the removal of gluten in beer was efficient.

Laser-induced fluorescence spectroscopy of plant-based drugs: Opium and hashish provoking at 405 nm.

Here, the fluorescence properties of some plant-based drug samples are characterized using a coherent excitation source at 405 nm. The laser-induced fluorescence (LIF) spectroscopy is examined to analyze opium and hashish. In order to improve traditional fluorescence methods for better analysis of optically dense materials, we have proposed five characteristic parameters based on solvent densitometry assay as the fingerprints of drugs of interest. The signal emissions are recorded in terms of various drug concentrations, such that the best fitting over experimental data determines the fluorescence extinction (α) and self-quenching (k) coefficients according to the modified Beer-Lambert formalism. The typical α value is determined to be 0.30 and 0.15 mL/(cm∙mg) for opium and hashish, respectively. Similarly, typical k is obtained 0.390 and 1.25 mL/(cm∙mg), respectively. Furthermore, the concentration at max fluorescence intensity (Cp) is determined for opium and hashish to be 1.8 and 1.3 mg/mL, respectively. Results reveal that opium and hashish benefit their own characteristic fluorescence parameters to discriminate those illicit substances promptly using the present method.

Impact of the Addition of Fruits of Kamchatka Berries (L. caerulea var. kamtschatica) and Haskap (L. caerulea var. emphyllocalyx) on the Physicochemical Properties, Polyphenolic Content, Antioxidant Activity and Sensory Evaluation Craft Wheat Beers.

Kamchatka berry (Lonicera caerulea var. kamtschatica) and haskap (Lonicera caerulea var. emphyllocalyx) fruit are important sources of bioactive compounds, mainly polyphenols, but also macro- and microelements. Physico-chemical analysis showed that wheat beers with added fruit were characterised by an average 14.06% higher ethanol content, lower bitterness and intense colour compared to the control, which was a wheat beer without added fruit. Wheat beers enriched with kamchatka berry fruit, including the "Aurora" variety, had the highest polyphenolic profile (e.g., chlorogenic acid content averaged 7.30 mg/L), and the antioxidant activity of fruit-enriched wheat beers determined by the DPPH method showed higher antioxidant activity of wheat beers enriched with kamchatka berry fruit, while those determined by the FRAP and ABTS methods showed higher antioxidant activity of wheat beers enriched with haskap fruit, including the "Willa" variety. Sensory evaluation of the beer product showed that wheat beers enriched with kamchatka berry fruits of the "Duet" variety and haskap fruits of the "Willa" variety were characterised by the most balanced taste and aroma. On the basis of the conducted research, it follows that both kamchatka berry fruits of the "Duet" and "Aurora" varieties and haskap fruit of the "Willa" variety can be used successfully in the production of fruity wheat beers.

Jambu Flower Extract (Acmella oleracea) Increases the Antioxidant Potential of Beer with a Reduced Alcohol Content.

Craft beers with different sensory perceptions have received the attention of more demanding consumers. In this sense, the application of plant extracts as brewing adjuncts is being increasingly studied. Allied with these perspectives is the consumption of lower alcoholic beverages, which also represents the desire for a market niche that has been growing gradually. The present work aimed to produce craft lager beer with the addition of plant extract and reduced alcohol content by partial replacement of malt with malt bagasse. The physical-chemical analyzes of the beer produced showed that it was possible to reduce the alcohol content by 40.5% compared to the control sample. In addition, an extract of Acmella oleracea (Jambu) obtained by supercritical extraction was added to increase the beer's antioxidant capacity. The ABTS, DPPH, and ORAC methods evaluated the antioxidant capacity. These assays were performed again after six months of storage. The quantification and identification of the significant substance in the extract (spilanthol) were performed using Gas Chromatography (GC-FID), Thin Layer Chromatography (TLC), and Attenuated Total Reflectance Infrared Spectroscopy (FTIR-ATR). The results showed significant increases in antioxidant activity compared to the sample without extract. This positive aspect opens a perspective for using jambu flower extract as a prominent antioxidant adjunct in beer.

The effect of CO2 concentration on yeast fermentation: rates, metabolic products, and yeast stress indicators.

This research aimed to assess how the partial removal of carbon dioxide affects fermentations to provide a better understanding of how the manipulation of carbon dioxide concentration can be used to optimize industrial fermentations. To achieve this, fermentation kinetics, fermentation metabolic products, and yeast stress indicators were analyzed throughout ongoing brewing fermentations conducted under partial vacuum with atmospheric pressure controls. The partial vacuum reduced the solubility of carbon dioxide in the media and decreased the time necessary to reach carbon dioxide saturation. The effect was an increased rate of fermentation, and significantly more viable cells produced under vacuum pressure compared to controls. Ethanol, glycerol, and volatile organic compound concentrations were all significantly increased under partial vacuum, while indicators of yeast stress (trehalose) were reduced. Additionally, as the number of yeast cells was higher under partial vacuum, less sugar was consumed per volume of yeast cell. This study measured fermentation kinetics, metabolic products, and yeast health to holistically assess the effect of partial vacuum during a batch fermentation and found significant differences in each that can be individually exploited by researchers and industry. SUMMARY: An exploration of batch yeast fermentation in a low-pressure environment, with a focus on the health and productivity of the yeast cells.

Evaluation of the antioxidant characteristics of craft beer with green tea.

The addition of green tea as antioxidants to beer can improve the beer's flavor stability by protecting against staling during storage. To analyze the effect of different green teas on the flavor stability of beer, we developed an approach to rapidly evaluate their antioxidant activity. Ten types of craft beer were produced by adding different kinds of green tea during brewing, and their antioxidant activity and phenolic profiles were evaluated. The results showed remarkable variations in antioxidant activity and antioxidative compound contents, which were considerably higher in green tea beers than in non-tea beer (p < 0.05). A comprehensive evaluation function was developed to evaluate the total antioxidant activity of beers using principal component analysis. The highest total antioxidant activity was observed in Taiping Houkui beer, with a comprehensive evaluation score of 2.53. Pearson correlation analysis suggested that (-)-epigallocatechin gallate, (-)-epicatechin gallate, and (-)-epigallocatechin were strongly correlated with the total antioxidant activity of green tea beers (p < 0.01). The summation of their contents represented more than 60% of the total phenolic content of the teas, which can be used to predict the total antioxidant activity of green tea beers. PRACTICAL APPLICATION: Flavor stability is of prime concern for brewers, and flavor aging is increasingly becoming the limiting factor in beer shelf life. The application of green tea as antioxidants in beer can improve the flavor stability by protecting against beer staling during storage. The analytical method developed in this study will contribute to the rapid comparison of the effect of different green teas on the flavor stability of beer. Furthermore, the research findings demonstrate the potential benefits of green teas to beer flavor stability, which is of considerable importance in promoting the development and consumption of green teas.

Beer-derived nitrogen, phosphorus co-doped carbon quantum dots: Highly selective on-off-on fluorescent probes for the detection of ascorbic acid in fruits.

A rapid, facile and ultrasensitive fluorescence sensing system based on beer-derived nitrogen, phosphorus co-doped carbon quantum dots (N, P-CQDs) for the detection of ascorbic acid (AA) in fruits was proposed. N, P-CQDs were successfully synthesized by one-step hydrothermal method, which afforded a high quantum yield (21.7 %), and showed the fluorescence with a maximum emission wavelength of 450 nm at an excitation wavelength of 370 nm. Further, N, P-CQDs were employed as an efficient sensor for ultrasensitive Fe3+-detection at concentrations ranging from 1-20 µM and 100-300 µM, respectively. N, P-CQDs@Fe3+ showed a high sensitivity and selectivity for AA detection. A linear response range for AA was obtained from 1 to 200 µM with limit of detection of 0.84 µM was obtained for AA. The result of MTT test showed that N, P-CQDs exhibit low toxicity, providing fast, accurate and less toxic route for testing AA in the food analysis fields.

Use of Coffee Bean Bagasse Extracts in the Brewing of Craft Beers: Optimization and Antioxidant Capacity.

Coffee bean bagasse is one of the main by-products generated by industrial coffee production. This by-product is rich in bioactive compounds such as caffeine, caffeic and chlorogenic acid, and other phenols. The aims of this work are to optimize the extraction conditions of phenolic compounds present in coffee bean bagasse and incorporate them into stout-style craft beers, as well as to determine their effect on the phenol content and antioxidant capacity. The optimal conditions for extraction were 30% ethanol, 30 °C temperature, 17.5 mL of solvent per gram of dry sample, and 30 min of sonication time. These conditions presented a total phenol content of 115.42 ± 1.04 mg GAE/g dry weight (DW), in addition to an antioxidant capacity of 39.64 ± 2.65 µMol TE/g DW in DPPH• and 55.51 ± 6.66 µMol TE/g DW for FRAP. Caffeine, caffeic and chlorogenic acids, and other minor compounds were quantified using HPLC-DAD. The coffee bean bagasse extracts were added to the stout craft beer and increased the concentration of phenolic compounds and antioxidant capacity of the beer. This work is the first report of the use of this by-product added to beers.

Glycemic indices and effect of bitter leaf (Vernonia amygdalina) flavored non-alcoholic wheat beer (NAWB) on key carbohydrate metabolizing enzymes in high fat diet fed (HFD)/STZ-induced diabetic Wistar rats.

In a bid to make the use of functional food easier in the management and prevention of diseases, product development and fortification from/with functional foods have become the recent focus of research. This study, therefore, sought to exploit the recent trend in the brewing industry on the production of non-alcoholic beers by investigating the possibility of having a non-alcoholic beer flavored with bitter leaf, a known plant widely reported to have a strong hypoglycemic effect, as against the traditional use of hops, and the effect of the produced beer on the glycemic indices and various diabetic biochemical parameters that serve as biomarkers for type-2 diabetes (T2D). The glycemic indices, as well as the inhibitory potentials of bitter leaf-flavored Non-alcoholic wheat beer (NAWB) in ratios of 100%HP, 100%BL, 75:25BL, 50:50BL, and 25:75BL, on enzymes linked to a high-fat diet/streptozocin (HFD/STZ)-induced T2D albino Wistar rats were investigated. There were no significant difference (p > .05) between the starch (1.72-1.77 mg/100 mL), amylose (0.22-0.24 mg/100 mL), and amylopectin (1.49-1.53 mg/100 mL) contents of the various samples. The Glycemic Index (GI) of the samples ranged from 36 to 73 with 75:25Bl and 50:50BL have the lowest (36) values. The samples reduced blood glucose levels and inhibited pancreatic α-amylase, lipase, and intestinal α-glucosidase activity. The inhibitory potentials of these beer samples on α-amylase and α-glucosidase as well as their ability to reduce blood glucose levels in diabetic rats thus making the bitter leaf flavored NAWB a suitable healthy beverage for better glycemic control in diabetics. PRACTICAL APPLICATIONS: This study revealed the potential of producing non-alcoholic wheat beer flavored with bitter leaves as a possible substitute for hops. The potential inherent in bitter leaf in the management of type 2 diabetes can thus be made available through a far-reaching beverage medium such as non-alcoholic beer to help in the treatment/management of T2D. The results of this research could be an eye-opener to the possible utilization of bitter leaf and by extension other plants that have been reported in the management of T2D. The use of the bitter leaf as a substitute for hops in the production of non-alcoholic beer in the brewing industry could help in a health-oriented campaign for safe drinks that could be helpful in the control of blood glucose levels of diabetic patients.

The Potent Phytoestrogen 8-Prenylnaringenin: A Friend or a Foe?

8-prenylnaringenin (8-PN) is a prenylated flavonoid, occurring, in particular, in hop, but also in other plants. It has proven to be one of the most potent phytoestrogens in vitro known to date, and in the past 20 years, research has unveiled new effects triggered by it in biological systems. These findings have aroused the hopes, expectations, and enthusiasm of a "wonder-drug" for a host of human diseases. However, the majority of 8-PN effects require such high concentrations that they cannot be reached by normal dietary exposure, only pharmacologically; thus, adverse impacts may also emerge. Here, we provide a comprehensive and up-to-date review on this fascinating compound, with special reference to the range of beneficial and untoward health consequences that may ensue from exposure to it.

Assessment of brewing attitude of unmalted cereals and pseudocereals for gluten free beer production.

In this study, 40% of unmalted gluten free (GF) grains (sorghum, millet, buckwheat, quinoa and amaranth) was used in brewing process, in gelatinized and ungelatinized form, in order to produce GF beer and to extend current knowledge about their suitability as brewing adjuncts. Partial replacement of barley malt with GF grains led to a significant decrease of extract (°P) and alcohol (%v/v) content compared to control beer (p < 0.05), except for quinoa beer (QB). Results from Principal Component Analysis (PCA) highlighted a satisfactory classification of experimental beers according to the two different forms of GF grains (gelatinized and ungelatinized). However, beers brewed with ungelatinized grains (mainly sorghum and quinoa) showed acceptable technological and sensory properties, thus suggesting that the pre-gelatinization step could be bypassed with a view to more environmentally and economically sustainable time-saving process. In addition, all beer samples showed a gluten content higher than 20 ppm.

Progress of the use of alternatives to malt in the production of gluten-free beer.

Beer is the most widely consumed alcoholic drink in the world, but it is not suitable for patients who suffer from celiac disease (CD) because its main ingresdients, barley or wheat, contain gluten. Approximately 1% of the world's population is affected by CD, and the development of gluten-free beer is imperative. Gluten-free beers produced using alternative materials, such as rice, sorghum, maize, millet, oats, and pseudocereals (e.g., buckwheat, quinoa and Amaranth), are studied in this review that examines the effects of specific substitutions on the different characteristics of the final beer to ensure the appropriateness of their use. The use of alternatives to malt may affect the quality of gluten-free beer and result in some negative consequences. Accordingly, the influential factors are discussed in terms of the total substitution of malt with other grains in the production of beer. Research results have provided some new alternative solutions for the production of gluten-free beer, such as the use of malted grains to improve hydrolytic enzyme activity, the application of nonconventional mashing procedures involving the decoction method and extrusion cooking techniques to increase the extract yield, the use of exogenous enzymes and nitrogen supplements to improve the sugar and amino acid spectra necessary for yeast fermentation, and the application of combinations of alternative grains to improve the flavor, body and foam stability of gluten-free beers.

Selenium inorganic speciation in beers using MSFIA-HG-AFS system after multivariate optimization.

In this work, the use of a multisyringe flow injection analysis coupled to hydride generation atomic fluorescence spectrometry (MSFIA-HG-AFS) for inorganic selenium chemical speciation was proposed. A Doehlert design was applied to optimize the experimental conditions for hydride generation (NaBH4 and HCl concentrations). The limits of quantification (LoQ) obtained were 0.07 µg L-1, for total inorganic Se, and 0.08 µg L-1, for Se(IV). Accuracy and precision of the proposed analytical method were evaluated through analysis of standard reference material and addition and recovery tests. The optimized method was applied to analyses of eight samples of beer, produced in Spain, obtaining concentrations for Se(IV) (<0.08 - 0.46 ± 0.01 µg L-1), total inorganic Se (0.47 ± 0.01 - 3.04 ± 0.62 µg L-1) and Se(VI) (0.06 ± 0.01 - 3.00 ± 0.59 µg L-1). The proposed analytical method was accurate, precise and sensitivity for determination of selenium species in beer samples.

Identification of in vitro angiotensin-converting enzyme and dipeptidyl peptidase IV inhibitory peptides from draft beer by virtual screening and molecular docking.

BACKGROUND: Hypertension and diabetes are two kinds of senile diseases which often occur simultaneously. The commonly used drugs in clinic may produce certain side effects. Food-derived polypeptide is a kind of polypeptide with great development potential, which has many functions of regulating human physiological function. Beer is rich in nutrition but there are few researches on bioactive peptides in beer. RESULTS: In this study, a rapid virtual screening method was established to obtain bioactive peptides from Tsingtao draft beer. The peptide sequence was analyzed by ultra-performance liquid chromatography-quadrupole-Orbitrap-tandem mass spectrometry (UPLC-Q-Orbitrap-MS2 ), and 50 peptides were identified. Eight peptides with potential biological activities were screened by using Peptide Ranker software and previous literature references. On the basis of absorption prediction, toxicity prediction, and molecular docking analysis, LNFDPNR and LPQQQAQFK were finally confirmed. The molecular docking results showed that two peptides could bind angiotensin-converting enzyme (ACE) and dipeptidyl peptidase IV (DPP-IV) tightly by hydrogen bonding and hydrophobic interaction. The in vitro activity evaluation results showed that two peptides had obvious ACE and DPP-IV inhibitory activity. CONCLUSION: This study established a method for rapidly screening bioactive peptides from Tsingtao draft beer, screened two ACE and DPP-IV inhibitory peptides in beer and analyzed their active action mechanism. This article may have great theoretical significance and practical value to further explore the health function of beer. © 2021 Society of Chemical Industry.

Hallstatt miners consumed blue cheese and beer during the Iron Age and retained a non-Westernized gut microbiome until the Baroque period.

We subjected human paleofeces dating from the Bronze Age to the Baroque period (18th century AD) to in-depth microscopic, metagenomic, and proteomic analyses. The paleofeces were preserved in the underground salt mines of the UNESCO World Heritage site of Hallstatt in Austria. This allowed us to reconstruct the diet of the former population and gain insights into their ancient gut microbiome composition. Our dietary survey identified bran and glumes of different cereals as some of the most prevalent plant fragments. This highly fibrous, carbohydrate-rich diet was supplemented with proteins from broad beans and occasionally with fruits, nuts, or animal food products. Due to these traditional dietary habits, all ancient miners up to the Baroque period have gut microbiome structures akin to modern non-Westernized individuals whose diets are also mainly composed of unprocessed foods and fresh fruits and vegetables. This may indicate a shift in the gut community composition of modern Westernized populations due to quite recent dietary and lifestyle changes. When we extended our microbial survey to fungi present in the paleofeces, in one of the Iron Age samples, we observed a high abundance of Penicillium roqueforti and Saccharomyces cerevisiae DNA. Genome-wide analysis indicates that both fungi were involved in food fermentation and provides the first molecular evidence for blue cheese and beer consumption in Iron Age Europe.

Redox and Anti-Inflammatory Properties from Hop Components in Beer-Related to Neuroprotection.

Beer is a fermented beverage widely consumed worldwide with high nutritional and biological value due to its bioactive components. It has been described that both alcoholic and non-alcoholic beer have several nutrients derived from their ingredients including vitamins, minerals, proteins, carbohydrates, and antioxidants that make beer a potential functional supplement. Some of these compounds possess redox, anti-inflammatory and anticarcinogenic properties making the benefits of moderate beer consumption an attractive way to improve human health. Specifically, the hop cones used for beer brewing provide essential oils, bitter acids and flavonoids that are potent antioxidants and immune response modulators. This review focuses on the redox and anti-inflammatory properties of hop derivatives and summarizes the current knowledge of their neuroprotective effects.

Impact of copper-based fungicides on the antioxidant quality of ethanolic hop extracts.

Hops contain a variety of compounds possessing antioxidant capacity including phenolic and polyphenolic compounds as well as α- and ß- acids. These compounds may contribute to the oxidative stability of beer during brewing and storage. Hop plants may be treated with copper-based fungicides (CBFs) which have been shown to increase the total copper content of harvested hop cones; however, copper ions are well known to catalyze the generation of reactive oxygen species production in beer and may negatively impact its oxidative stability. Increased copper content in CBF-treated hops has been previously shown to have deleterious effects on the aroma quality of hops and beer. The impact of CBFs on the antioxidant content and quality of hops has not been previously investigated. In this study, ethanolic extracts of CBF-treated hops are evaluated for their metal content and antioxidant quality in order to determine whether excess copper from CBF treatment negatively impacts their antioxidant capacity.

Effects of the intake of craft or industrial beer on serum homocysteine.

Beer is a source of folate, vitamin B6 and B12, molecules involved in the pathways of homocysteine (HCY), a risk factor for cardiovascular disease. This research evaluated if a consumption of craft or industrial beer could reduce serum HCY. In a randomised cross-over study, 12 men (28.7 ± 6.0 years) and 12 women (29.4 ± 7.5 years), healthy, omnivorous, with normal body mass index, non-smoking and not taking oral supplements or contraceptives, followed a free-living diet and received, daily, for 3 weeks, 330 ml of industrial (4.5% of alcohol) or craft beer (9% of alcohol). Anthropometric measures and blood samples were taken at the beginning and at the end of each period. The consumption of industrial beer reduced (p < 0.05) HCY (7.35 vs. 6.50 µmol/L) and increased folic acid (3.46 vs. 3.94 ng/mL). Craft beer increased gamma-gluamyl transpeptidase (GGT) (16.6 vs. 18.6 U/L) and reduced vitamin B6 (20.9 vs. 16.9 ng/mL).

Flavonoids as Phytoestrogenic Components of Hops and Beer.

The value of hops (Humulus lupulus L.) in beer production has been undisputed for centuries. Hops is rich in humulones and lupulones which gives the characteristic aroma and bitter taste, and preserves this golden drink against growing bacteria and molds. Besides α- and ß-acids, the lupulin glands of hop cones excrete prenylated flavonoids, which exhibit a broad spectrum of biological activities and therefore has therapeutic potential in humans. Recently, interest in hops was raised due to hop prenylated flavanones which show extraordinary estrogen activities. The strongest known phytoestrogen so far is 8-prenylnaringenin (8-PN), which along with 6-prenylanaringenin (6-PN), 6,8-diprenylnaringenin (6,8-DPN) and 8-geranylnaringenin (8-GN) are fundamental for the potent estrogen activity of hops. This review provides insight into the unusual hop phytoestrogens and shows numerous health benefits associated with their wide spectrum of biological activities including estrogenic, anticancer, neuropreventive, antinflamatory, and antimicrobial properties, which were intensively studied, and potential applications of these compounds such as, as an alternative to hormone replacement therapy (HRT).

Fractionated stir bar sorptive extraction using conventional and solvent-assisted approaches for enhanced identification capabilities of aroma compounds in beverages.

For successful profiling of aroma carriers in food samples, a highly efficient extraction method is mandatory. A two-step stir bar sorptive extraction (SBSE) approach, namely fractionated SBSE (Fr-SBSE), was developed to improve both the organoleptic and the chemical identification of aroma compounds in beverages. Fr-SBSE consists of two multi-SBSE procedures (mSBSE) performed sequentially on the same sample. The first extraction consists of a conventional mSBSE using three polydimethylsiloxane (PDMS) stir bars (1stmSBSE). This is followed by a solvent-assisted mSBSE performed on the same sample using three solvent-swollen PDMS stir bars (2nd SA-mSBSE). The 1stmSBSE mainly extracts apolar/medium polar solutes with log Kow values >2, while the 2nd SA-mSBSE mainly extracts polar solutes with log Kow values <2. After this two-step fractionation procedure, either thermal desorption (TD) or liquid desorption - large volume injection (LD-LVI), followed by GC-MS is performed on each set of three stir bars. A real-life sample of roasted green tea was used for method development. The performance of the Fr-SBSE method is further illustrated with sensory evaluations and GC-MS analysis for a stout beer sample. Compared to an extraction procedure with SA-mSBSE only, Fr-SBSE including a 1stmSBSE and a 2nd SA-mSBSE reduced co-elution of aroma compounds in the chromatograms and was capable of providing improved mass spectral quality for identification of 17 additional compounds in roasted green tea, and 12 additional compounds in stout beer, respectively. Moreover, odor description and characterization were clearly improved.