Thermodynamics of semi-specific ligand recognition: the binding of dipeptides to the E.coli dipeptide binding protein DppA.

This investigation of the temperature dependence of DppA interactions with a subset of three dipeptides (AA. AF and FA) by isothermal titration calorimetry has revealed the negative heat capacity ([Formula: see text]) that is a characteristic of hydrophobic interactions. The observation of enthalpy-entropy compensation is interpreted in terms of the increased structuring of water molecules trapped in a hydrophobic environment, the enthalpic energy gain from which is automatically countered by the entropy decrease associated with consequent loss of water structure flexibility. Specificity for dipeptides stems from appropriate spacing of designated DppA aspartate and arginine residues for electrostatic interaction with the terminal amino and carboxyl groups of a dipeptide, after which the binding pocket closes to become completely isolated from the aqueous environment. Any differences in chemical reactivity of the dipeptide sidechains are thereby modulated by their occurrence in a hydrophobic environment where changes in the structural state of entrapped water molecules give rise to the phenomenon of enthalpy-entropy compensation. The consequent minimization of differences in the value of ΔG0 for all DppA-dipeptide interactions thus provides thermodynamic insight into the biological role of DppA as a transporter of all dipeptides across the periplasmic membrane.

Effects of aroma and taste, independently or in combination, on appetite sensation and subsequent food intake.

Food flavour is important in appetite control. The effects of aroma and taste, independently or in combination, on appetite sensation and subsequent food intake, were studied. Twenty-six females (24 ± 4 years, 20.9 ± 1.9 kg⋅m-2) consumed, over 15 min period, one of four sample drinks as a preload, followed by an ad libitum consumption of a pasta meal (after 65 min). Sample drinks were: water (S1, 0 kcal), water with strawberry aroma (S2, 0 kcal), water with sucrose and citric acid (S3, 48 kcal) and water with strawberry aroma, sucrose and citric acid (S4, 48 kcal). Appetite sensation did not differ between the S1 (water), S2 (aroma) and S3 (taste) conditions. Compared with S1 (water), S2 (aroma) and S3 (taste), S4 (aroma + taste) suppressed hunger sensation over the 15 min sample drink consumption period (satiation) (p < 0.05). S4 (aroma + taste) further reduced hunger sensation (satiety) more than S1 at 5, 20 and 30 min after the drink was consumed (p < 0.05), more than S2 (aroma) at 5 and 20 min after the drink was consumed (p < 0.05), and more than S3 (taste) at 5 min after the drink was consumed (p < 0.05). Subsequent pasta energy intake did not vary between the sample drink conditions. S4 (aroma + taste) had the strongest perceived flavour. This study suggests that the combination of aroma and taste induced greater satiation and short-term satiety than the independent aroma or taste and water, potentially via increasing the perceived flavour intensity or by enhancing the perceived flavour quality and complexity as a result of aroma-taste cross-modal perception.

Sharing a Host Plant (Wheat [Triticum aestivum]) Increases the Fitness of Fusarium graminearum and the Severity of Fusarium Head Blight but Reduces the Fitness of Grain Aphids (Sitobion avenae).

We hypothesized that interactions between fusarium head blight-causing pathogens and herbivores are likely to occur because they share wheat as a host plant. Our aim was to investigate the interactions between the grain aphid, Sitobion avenae, and Fusarium graminearum on wheat ears and the role that host volatile chemicals play in mediating interactions. Wheat ears were treated with aphids and F. graminearum inoculum, together or separately, and disease progress was monitored by visual assessment and by quantification of pathogen DNA and mycotoxins. Plants exposed to both aphids and F. graminearum inoculum showed accelerated disease progression, with a 2-fold increase in disease severity and 5-fold increase in mycotoxin accumulation over those of plants treated only with F. graminearum. Furthermore, the longer the period of aphid colonization of the host prior to inoculation with F. graminearum, the greater the amount of pathogen DNA that accumulated. Headspace samples of plant volatiles were collected for use in aphid olfactometer assays and were analyzed by gas chromatography-mass spectrometry (GC-MS) and GC-coupled electroantennography. Disease-induced plant volatiles were repellent to aphids, and 2-pentadecanone was the key semiochemical underpinning the repellent effect. We measured aphid survival and fecundity on infected wheat ears and found that both were markedly reduced on infected ears. Thus, interactions between F. graminearum and grain aphids on wheat ears benefit the pathogen at the expense of the pest. Our findings have important consequences for disease epidemiology, because we show increased spread and development of host disease, together with greater disease severity and greater accumulation of pathogen DNA and mycotoxin, when aphids are present.

Comparison of ambient solvent extraction methods for the analysis of fatty acids in non-starch lipids of flour and starch.

BACKGROUND: Lipids are minor components of flours, but are major determinants of baking properties and end-product quality. To the best of our knowledge, there is no single solvent system currently known that efficiently extracts all non-starch lipids from all flours without the risk of chemical, mechanical or thermal damage. This paper compares nine ambient solvent systems (monophasic and biphasic) with varying polarities: Bligh and Dyer (BD); modified Bligh and Dyer using HCl (BDHCL); modified BD using NaCl (BDNaCl); methanol-chloroform-hexane (3:2:1, v/v); Hara and Radin (hexane-isopropanol, 3:2, v/v); water-saturated n-butanol; chloroform; methanol and hexane for their ability to extract total non-starch lipids (separated by lipid classes) from wheat flour (Triticum aestivum L.). Seven ambient extraction protocols were further compared for their ability to extract total non-starch lipids from three alternative samples: barley flour (Hordeum vulgare L.), maize starch (Zea mays L.) and tapioca starch (Manihot esculenta Crantz). RESULTS: For wheat flour the original BD method and those containing HCl or NaCl tended to extract the maximum lipid and a significant correlation between lipid extraction yield (especially the glycolipids and phospholipids) and the polarity of the solvent was observed. For the wider range of samples BD and BD HCl repeatedly offered the maximum extraction yield and using pooled standardized (by sample) data from all flours, total non-starch lipid extraction yield was positively correlated with solvent polarity (r = 0.5682, P < 0.05) and water ratio in the solvent mixture (r = 0.5299, P < 0.05). CONCLUSION: In general, BD-based methods showed better extraction yields compared to methods without the addition of water and, most interestingly, there was much greater method dependence of lipid yields in the starches when compared to the flour samples, which is due to the differences in lipid profiles between the two sample types (flours and starches).

Entrapment of a volatile lipophilic aroma compound (d-limonene) in spray dried water-washed oil bodies naturally derived from sunflower seeds (Helianthus annus).

Oil bodies are natural emulsions that can be extracted from oil seeds and have previously been shown to be stable after spray drying. The aim of the study was to evaluate for the first time if spray dried water-washed oil bodies are an effective carrier for volatile lipophilic actives (the flavour compound d-limonene was used as an example aroma compound). Water-washed oil bodies were blended with maltodextrin and d-limonene and spray dried using a Buchi B-191 laboratory spray dryer. Lipid and d-limonene retention was 89-93% and 24-27%. Samples were compared to processed emulsions containing sunflower oil and d-limonene and stabilised by either lecithin or Capsul. Lecithin and Capsul processed emulsions had a lipid and d-limonene retention of 82-89%, 7.7-9.1% and 48-50%, 55-59% respectively indicating that water-washed oil bodies could retain the most lipids and Capsul could retain the most d-limonene. This indicates that whilst additional emulsifiers may be required for future applications of water-washed oil bodies as carriers of lipophilic actives, oil bodies are excellent agents for lipid encapsulation.

Effect of sweeteners and carbonation on aroma partitioning and release in beverage systems.

The effect of monosaccharides (glucose, fructose and galactose) and disaccharides (sucrose and lactose) at 10, 20 and 30 % w/v on the in-vitro aroma partitioning of C4 - C10 aldehydes and ethyl esters, as well as limonene (concentration of aroma compounds at 1 µg mL-1), was studied using atmospheric pressure chemical ionisation-mass spectrometry. An increase in sugar concentration from 0 to 30 % w/v resulted in a significant increase in partitioning under static headspace conditions for the majority of the compounds (p < 0.05), an effect generally not observed when 10 % w/v sucrose was substituted with low-calorie sweeteners (p > 0.05). The complexity of the system was increased to model a soft drink design - comprising water, sucrose (10, 20 and 30 % w/v), acid (0.15 % w/v), carbonation (∼7.2 g/L CO2) and aroma compounds representative of an apple style flavouring, namely ethyl butanoate and hexanal (10 µg mL-1 each). Although the addition of sucrose had no significant in-vivo effect, carbonation significantly decreased breath-by-breath (in-vivo) aroma delivery (p < 0.05). To understand the physical mechanisms behind aroma release from the beverage matrix, the effect of sucrose on the kinetics of the matrix components was explored. An increase in sucrose concentration from 0 to 30 % w/v resulted in a significant decrease in water activity (p < 0.05), which accounted for the significantly slower rate of self-diffusion of aroma compounds (p < 0.05), measured using diffusion-ordered spectroscopy-nuclear magnetic resonance spectroscopy. No significant effect of sucrose on carbon dioxide volume flux was found (p > 0.05).

The role of inorganic-phosphate, potassium and magnesium in yeast-flavour formation.

Inorganic-phosphate, potassium, and magnesium are key-minerals required for yeast growth, metabolism, and survival, the present work investigated its impact in yeast-flavour formation using a multi-factor experimental design, which was used to generate a range of phosphorous-potassium-magnesium resulting in a 28-point D-optimal design. Samples were evaluated using HPLC (ethanol), GC-MS (aroma), and CountStar (total yeast cell). Results revealed that yeast requires a minimal amount of inorganic-phosphate, potassium, and magnesium (250, 500, and 70 mg/L, respectively) to support yeast-growth and ethanol/flavour formation. Inorganic-phosphate was important for fatty acid esters formation/short chain fatty acid (SCFA) reduction. Potassium was important in the formation of acetate esters/higher alcohols. Magnesium was the most important inorganic element for ester formation/SCFA reduction; furthermore, ethanol production is magnesium-dependent. In conclusion, inorganic phosphate, potassium and magnesium play an important role in yeast-growth, esters and higher alcohols formation; and SCFA reduction. Ethanol formation is Mg-dependent.

Influence of essential inorganic elements on flavour formation during yeast fermentation.

The relative concentration of available inorganic elements is critical for yeast growth and metabolism and has potential to be a tool leading to directed yeast flavour formation during fermentation. This study investigates the influence of essential inorganic elements during alcoholic fermentation of brewers wort, fermented using three independent yeast strains, Saccharomyces pastorianus W34/70, and Saccharomyces cerevisiae strains M2 and NCYC2592 under a range of conditions replicated for each yeast strain. 10 treatments were applied: 1 control and 9 inorganic supplementations: standard brewers wort, ammonia-nitrogen, inorganic phosphate, potassium, magnesium, copper, zinc, iron, manganese and a composite mixture, Twenty-five chemical markers were evaluated by HPLC (ethanol, glycerol), and GC-MS (aroma). There was a significant change in volatile aroma compounds during fermentation, which was more prominent when supplemented with ammonia nitrogen, inorganic phosphate, potassium or magnesium (P < 0.05). Heavy metal ions mostly had a negative effect on the flavour formation.

Reducing sugar and aroma in a confectionery gel without compromising flavour through addition of air inclusions.

Sugar plays an important role in both the flavour and structure of confectionery. Targets have been set to reduce sugar; however, common strategies often result in changes in flavour and consumer rejection. In this study, an approach was developed to reduce sugar in confectionery gels by aeration, without significantly affecting perceived chewiness. Gelatine based gels with a 23% and 38% reduction in density were formulated using aeration. Mean bubble size was consistent across all gels (0.05-0.06 mm). Time-intensity sensory evaluation was carried out by a trained sensory panel (n = 10). With aeration, no significant difference in overall flavour perception was observed in the 23% and 38% reduced sugar and aroma gels. Air inclusions create a greater surface area, therefore accelerating mass transfer of volatiles and release of sucrose at the food-saliva interface. Consequently, we propose that less calorie dense products can be produced without compromising flavour by using gel aeration technologies.

The role of sodium chloride in the sensory and physico-chemical properties of sweet biscuits.

Salt is included in many foods which consumers do not regard as salty. This "hidden-salt" may offer functional benefits but is often overlooked in sodium reduction strategies. This study investigated its role in shortbread-like sweet biscuits (1.05 g NaCl/100 g). Sensory tests revealed significant flavour and texture differences after a salt reduction of 33% (0.86 g/ 100 g). This was explained by differences in the partitioning of hydrophobic aroma compounds into the headspace and a significant impact on structure. Texture analysis and X-ray-µCT measurements revealed a reduced hardness with larger and more air cells in salt-reduced biscuits. It is suggested that salt impacts on cereal proteins by altering their aggregation around flour particles and at bubble walls and that slower water loss occurs in salted matrices during baking. Hence, this study revealed the key properties significantly affected by salt reduction and proposes an explanation which will help to develop a targeted "hidden-salt" reduction strategy.

A comprehensive investigation of gluten free bread dough rheology, proving and baking performance and bread qualities by response surface design and principal component analysis.

Contribution of methylcellulose (MC), psyllium seed husk powder (PSY), and water addition level to gluten free bread quality and correlations between dough rheological properties and bread qualities were investigated by response surface design and principal component analysis. The generalised Maxwell model was applied to estimate the relaxation frequency of gluten free doughs. The addition of PSY has a complex influence on pasting viscosity at high temperature and an additional peak was observed. MC significantly influenced dough extensibility and work of adhesion, which are good predictors of bread volume and textural properties. Other rheological responses are less significantly correlated to specific volume, but they are sensitive to formulation variations, reflect dough structures and stability, related to proving behaviours, and correlated to loaf concavity. An inappropriate combination of water and hydrocolloids might lead to problems such as low stability of doughs, overexpansion, and weak crumb structure at high water addition levels, or, in contrast, high rigidity of dough, a trap of excessive air during mixing, and restrained gas cell expansion with high hydrocolloid addition and low water addition.

Characterisation of high molecular weight hop proanthocyanidins using Analytical Ultracentrifugation.

We report the novel application of Analytical Ultracentrifugation (AUCF) to characterise the polymeric proanthocyanidin fraction of hops. Extraction of hop samples with 70% acetone (aq) followed by a C-18 Solid Phase Extraction yielded polyphenolic fractions for AUCF analysis. Sedimentation velocity experiments demonstrated the presence of discrete molecular weight bands of proanthocyanidins, as opposed to a continuous distribution of molecular weights. There were 4 such bands for Saaz hop (0.15, 1.1, 2.7 and 4.4S) and 3 bands for Magnum (0.15, 1.6 and 3.0S). The method resulted in a reproducible size (sedimentation coefficient) distribution for replicate runs of the same extract and for extracts prepared from different samples of the same hop variety. Sedimentation equilibrium experiments were then used to fit molecular weight distributions using the new SEDFIT-MSTAR method for the same samples. Thus we report for the first time polymeric proanthocyanidins in hops with molecular weights of up to 100 kDa in Saaz hop (or up to 56 kDa in Magnum). This represents the first application of AUCF to characterise complex fractions of polyphenolics extracted from botanical sources and the methodology developed should find wider application in the study of this diverse and bioactive class of compounds.

Dynamic Aroma Release from Complex Food Emulsions.

In vitro dynamic aroma release over oil-in-water (o/w) and water-in-oil-in-water (w/o/w) emulsions stabilized with Tween 20 or octenyl succinic anhydride (OSA) starch as a hydrophilic emulsifier and polyglycerol polyricinoleate (PGPR) as a hydrophobic emulsifier was investigated. The equal-molecular-weight hydrophilic aroma diacetyl (2,3-butanedione) or relatively more-hydrophobic 3-pentanone was added to the emulsions prepared by high speed mixing, or membrane emulsification followed by thickened with xanthan gum removing droplet size distribution and creaming as variables affecting dynamic release. Results showed the differences of w/o/w emulsions in the dynamic release compared to o/w emulsions mainly depended on aroma hydrophobicity, emulsion type, emulsifier-aroma interactions, and creaming. Xanthan led to a reduced headspace replenishment. Interfacially adsorbed OSA starch and xanthan-OSA starch interaction influenced on diacetyl release over emulsions. OSA starch alone interacted with 3-pentanone. This study demonstrates the potential impact of emulsifying and thickening systems on aroma release systems and highlights that specific interactions may compromise product quality.

Acid production, growth kinetics and aroma profiles of Lactobacillus flora from Stilton cheese.

The effect of Lactobacillus plantarum isolates from Stilton cheese on aroma profiles of milk fermentation was examined. Representative Lb. plantarum isolates were cultured alone and in combination with acid-producing and non-acid producing Lactococcus lactis NCIMB 9918 in UHT milk at 30 & 18 °C for 48 h & 12 weeks, respectively in presence and absence of salt, simulating cheese production and ripening. During long-term ripening, Lb. plantarum grew faster when co-cultured with non-acid producing Lc. lactis in the presence of salt. One isolate of Lb. plantarum produced the highest concentration of alcohols, organic acids and acetoin. Co-culture of Lb. plantarum with acid-producing Lc. lactis enhanced acid and alcohol production, whereas co-inoculation with non-acid producing Lc. lactis increased acetoin synthesis. Lb. plantarum is an incidental organism in cheese and its presence is unpredictable. Occurrence of different genotypes of Lb. plantarum could contribute to batch to batch variation in the cheese aroma characteristics.

Modifying Robusta coffee aroma by green bean chemical pre-treatment.

Green Robusta beans were subjected to pre-treatment with the aim of reducing the perceived aroma difference between Arabica and Robusta coffee. Treatment was a short soaking procedure with varying concentrations of acetic acid (up to 5%). Samples were subjected to thermal treatment (roasted) and ground to a standardised particle size distribution. Aroma compounds were evaluated by headspace analysis using solid-phase microextraction and gas chromatography-mass spectrometry. Pre-treatment significantly affected aroma formation during roasting and resulted in a modified level of pyrazines, furanic compounds and sulfur-containing compounds (p < 0.05). Principal component analysis illustrated that the aroma profile of the pre-treated Robusta coffee was closer to the target Arabica coffee after roasting. Sensory results confirmed that the aroma of the 2% acetic acid pre-treated Robusta brew was similar to Arabica; the maximum inclusion level of Robusta coffee in a blend could be increased from 20% to 80%.

Enhancing Robusta coffee aroma by modifying flavour precursors in the green coffee bean.

This study attempted to improve Robusta sensory properties by modifying the beans chemical composition. Building on our previous work, which modified bean pH through acid pre-treatment, a model system was developed where, sugar solutions (glucose, fructose, sucrose) were used to pre-treat Robusta coffee beans with the aim to modify the concentration/availability/location of these aroma precursors. Beans were then dried to equal water activity, subjected to equal roast intensity and ground to comparable particle size distributions. The treatment significantly impacted aroma generation during roasting leading to an altered level of pyrazines, furans, ketones, organic acid and heterocyclic nitrogen-containing compounds (p < 0.05). The optimum treatment was 15 g/100 g fructose. 80% treated Robusta could be blended with Arabica in coffee brew without significant aroma differences being perceived when compared to 100% Arabica brew. Furthermore the aroma of the fructose treated Robusta was more stable than Arabica over six weeks accelerated shelflife storage.

Aroma binding and stability in brewed coffee: A case study of 2-furfurylthiol.

The aroma stability of fresh coffee brew was investigated during storage over 60 min, there was a substantial reduction in available 2-furfurylthiol (2-FFT) (84%), methanethiol (72%), 3-methyl-1H-pyrole (68%) and an increase of 2-pentylfuran (65%). It is proposed that 2-FFT was reduced through reversible chemical binding and irreversible losses. Bound 2-FFT was released after cysteine addition, thereby demonstrating that a reversible binding reaction was the dominant mechanism of 2-FFT loss in natural coffee brew. The reduction in available 2-FFT was investigated at different pH and temperatures. At high pH, the reversible binding of 2-FFT was shown to protect 2-FFT from irreversible losses, while irreversible losses led to the reduction of total 2-FFT at low pH. A model reaction system was developed and a potential conjugate, hydroxyhydroquinone, was reacted with 2-FFT. Hydroxyhydroquinone also showed 2-FFT was released after cysteine addition at high pH.

Carbohydrate utilization and the lager yeast transcriptome during brewery fermentation.

The fermentable carbohydrate composition of wort and the manner in which it is utilized by yeast during brewery fermentation have a direct influence on fermentation efficiency and quality of the final product. In this study the response of a brewing yeast strain to changes in wort fermentable carbohydrate concentration and composition during full-scale (3275 hl) brewery fermentation was investigated by measuring transcriptome changes with the aid of oligonucleotide-based DNA arrays. Up to 74% of the detectable genes showed a significant (p

Effects of co-inoculation and sequential inoculation of Tetragenococcus halophilus and Zygosaccharomyces rouxii on soy sauce fermentation.

The use of Tetragenococcus halophilus and Zygosaccharomyces rouxii as starter cultures is essential for desirable volatiles production during moromi stage of soy sauce fermentation. In this study, the effect of simultaneous and sequential inoculation of cultures in moromi fermentation models, with respect to viability, physicochemical changes, and volatiles formation (using SPME-GC/MS) was investigated. Interestingly, an antagonism was observed as T. halophilus only proliferated (3 log increase) in the presence of Z. rouxii, while Z. rouxii growth was suppressed by 4 log in concurrence with pH increase to 7.31. Final content of reducing sugars, ethanol, acetic acid, and amino nitrogen did not differ significantly (p<0.05) between co-inoculation and sequential inoculation. However, Z. rouxii promoted alcohols formation and produced a more complex aroma profile under suppression. According to Principal Component Analysis (PCA), the inoculation sequence (co-inoculation and sequential) has impacts on volatile compound profiles during moromi fermentation.

The role of phenolic compounds on olive oil aroma release.

In this study, atmospheric pressure chemical ionisation-mass spectrometry (APCI-MS) was successfully applied to understand the effect of phenolic compounds on the release of olive oil aroma compounds. Eight aroma compounds were monitored under in-vivo and in-vitro dynamic conditions in olive oil with and without the addition of virgin olive oil (VOO) biophenols. Three model olive oils (MOOs) were set up with identical volatile compounds concentrations using a refined olive oil (ROO). Phenolics were extracted from VOOs and were added to two MOOs in order to obtain two different concentrations of phenolic compounds (P+ = 354 mg kg-1; P++ = 593 mg kg-1). Another MOO was without VOO biophenols (P-). Phenolic compounds impacted both the intensity and time of aroma release. In the in-vivo study, 1-penten-3-one, trans-2-hexenal and esters had lower release in the presence of higher levels of biophenols after swallowing. In contrast, linalool and 1-hexanol had a greater release. The more hydrophobic compounds had a longer persistence in the breath than the hydrophilic compounds. VOO phenolics-proline-rich proteins complexes could explain the binding of aroma compounds and consequently their decrease during analysis and during organoleptic assessment of olive oil.