Aroma compounds generated from thermal reaction of l-ascorbic acid with l-cysteine

The reaction of l-ascorbic acid with l-cysteine in heated aqueous solution (141 ± 1 °C) at five different pH values (5.00, 6.00, 7.00, 8.00, or 9.00) for 2 h, resulted in the formation of a complex mixture of aroma volatiles. The volatile compounds generated were analysed by SPME–GC–MS. The results gave 43 aroma compounds. The reaction between l-ascorbic acid and l-cysteine led mainly to the formation of alicyclic sulphur compounds, thiophenes, thienothiophenes, thiophenones, thiazoles and pyrazines, most of which contain sulphur. Many of these volatiles had meaty flavour. The origin of many of the compounds was explained. The studies showed that thienothiophenes and thienones were formed mainly at acidic pH. In contrast, higher pH values could promote the production of thiophenes, thiazoles and pyrazines.     

Identification and characterization of a novel l-amino acid ligase from Photorhabdus luminescens subsp. laumondii TT01

l-Amino acid ligase catalyzes dipeptide synthesis from unprotected l-amino acids in an ATP-dependent manner. We recently identified a new member of l-amino acid ligase, the plu1440 protein, from Photorhabdus luminescens subsp. laumondii TT01 by in silico analysis. This protein was found to synthesize dipeptides containing l-asparagine at the N-terminus, which is a novel substrate specificity.     

Mechanism of formation of sulphur aroma compounds from l-ascorbic acid and l-cysteine during the Maillard reaction

The sulphur aroma compounds produced from a phosphate-buffered solution (pH 8) of l-cysteine and l-, l-[1-¹³C] or l-[4-¹³C] ascorbic acid, heated at 140 ± 2 °C for 2 h, were examined by headspace SPME in combination with GC-MS. MS data indicated that C-1 of l-ascorbic acid was not involved in the formation of sulphur aroma compounds. The sulphur aroma compounds formed by reaction of l-ascorbic acid with l-cysteine mainly contained thiophenes, thiazoles and sulphur-containing alicyclic compounds. Among these compounds, 1-butanethiol, diethyl disulphide, 5-ethyl-2-methylthiazole, cis and trans-3,5-dimethyl-1,2,4-trithiolane, thieno[2,3-b]thiophene, thieno[3,2-b]thiophene, cis and trans-3,5-diethyl-1,2,4-trithiolane, 1,2,5,6-tetrathiocane, 2-ethylthieno[2,3-b]thiophene, 2,4,6-trimethyl-1,3,5-trithiane and cyclic octaatomic sulphur (S₈) were formed solely by l-cysteine degradation, and the rest by reaction of l-ascorbic acid degradation products, such as hydroxybutanedione, butanedione, acetaldehyde, acetol, pyruvaldehyde and formaldehyde with l-cysteine or its degradation products, such as H₂S and NH₃. A new reaction pathway from l-ascorbic acid via its degradation products was proposed.     

Effect of l-ascorbic acid, sugar, pectin and freeze-thaw treatment on polyphenol content of frozen strawberries

The stability of phenolic compounds of three strawberry cultivars was evaluated for changes during prefreezing treatments, storage and various freezing and thawing conditions. Polyphenol content was determined by HPLC-DAD-FL. The sum of assayed polyphenolic (proanthocyanidin and monomeric flavan-3-ols, anthocyanins, ellagic acid, p-coumaric acid) represented 2858.4 mg/kg in Kent, 2893.1 mg/kg in Elkat, and 2438.0 mg/kg in Senga Sengana. After freezing, 4.5-33.6% of polyphenols were lost; protective effects of prefreezing treatments were seen on anthocyanidins and proanthocyanidins: ascorbic acid was the most effective pretreatment, allowing retention of 3.9-27.5% of anthocyanins, and almost total recovery (3.9-23.9%) when associated with liquid nitrogen freezing. Pectin and sugar only allowed retention of 3.0-25.1% and 5.5-25.4% of the antocyanins, respectively. Thawing of the strawberries in a microwave oven (instead of 20 h at 20 °C) had a further positive effect on retention of anthocyanins, proanthocyanins, (+)-catechin and ellagic acid.     

Enhancement of l(+)-lactic acid production using acid-adapted precultures of Rhizopus arrhizus in a bubble column reactor

88 g/L lactic acid was produced from waste potato starch (equivalent to 100 g/L glucose) in a bubble column reactor using appropriate acid-adapted precultures of Rhizopus arrhizus. Further experiment showed that repeated dilution of cultures caused the decrease of lactic acid concentration and productivity due to formation of large fungal pellets.     

Capsular and slime-polysaccharide production by Lactobacillus rhamnosus JAAS8 isolated from Chinese sauerkraut: Potential application in fermented milk products

Exopolysaccharides (EPSs) produced by lactic acid bacteria (LAB) play an important role in the improvement of the physical properties of fermented dairy products. To find EPS-producing LAB strains with potential industrial applications, a Lactobacillus rhamnosus strain, L. rhamnosus JAAS8, that is capable of producing two forms of EPS when grown in MRS broth or semi-defined medium with glucose as a carbon source was isolated and identified from Chinese sauerkraut. The capsular-polysaccharide (CPS) present surrounding the bacterial surface of L. rhamnosus JAAS8 was observed by both optical microscopy and transmission electron microscopy. The slime-polysaccharide (SPS) present in the growth medium was produced mainly during the exponential growth phase, while the CPS was produced only in fermentation. Monosaccharide analysis of the purified polysaccharide samples showed that the CPS was composed of galactose and N-acetylglucosamine in a molar ratio of 5:1, and the SPS was composed of galactose, glucose and N-acetylglucosamine in a molar ratio of 4:1:1. The use of L. rhamnosus JAAS8 could be considered for potential applications in the dairy industry to improve the rheological properties of fermented milk products by increasing their water-holding capacity and viscosity.     

Screening of d-amino acid oxidase inhibitor by a new multi-assay method

This paper describes an investigation of the inhibition of a d-amino acid oxidase (DAAO) activity in several kinds of food additives and fruit. To screen for an inhibitor of DAAO, the method employed checked for two kinds of indicators, namely, pyruvic acid (indicator-1) and hydrogen peroxide (indicator-2), both of which are formed by digestion by DAAO of d-alanine (the standard d-amino acid used in this study). For measurement of pyruvic acid, a reliable and authenticated method was employed: the presence of pyruvic acid was determined by converting it into a chromophore derivative by tagging with 2,4-dinitro phenylhydrazine (DNP), followed by measurement of absorbance at 445 nm. The pyruvic acid thus determined is referred to as “(colorimetric) indicator-1”. To measure hydrogen peroxide, a highly sensitive fluorogenic substrate, 3-(p-hydroxyphenyl)propionic acid (HPPA), was employed in the present study. Hydrogen peroxide was detected by measuring fluorescence intensity (Ex. = 320 nm, Em. = 405 nm) of 2,2′-dihydroxybiphenyl-5,5′-dipropionic acid (oxidize-HPPA), which is produced by horseradish peroxidase (HRP)-catalyzed oxidation of HPPA with hydrogen peroxide. The hydrogen peroxide thus determined is referred to as “(flourometric) indicator-2” After optimizing the multi-assay procedures, the inhibition of DAAO activity against d-alanine (used as the standard d-amino acid) in 11 kinds of food additives and five kinds of fruit was evaluated, first in terms of indicator-1. Then, two substrates, potassium sorbate and apple juice, that were screened by indicator-1, were further evaluated by indicator-2 to conform the inhibition of DAAO activity. This is the first demonstration of the inhibition of DAAO activity by potassium sorbate, which is widely used as a synthetic preservative in food and apple juice.     

Rapid HPLC-UV method for quantification of l-citrulline in watermelon and its potential role on smooth muscle relaxation markers

A rapid and versatile HPLC-UV method for the separation and quantification of l-citrulline from watermelon was developed for the first time. Freeze dried juice and rind of watermelon were extracted with acidic methanol and analysed by HPLC using 0.03 mM phosphoric acid as a mobile phase. The separation was performed using Zorbax SB-Aq, Synergi Hydro-RP and Gemini C-18 columns and l-citrulline was detected at 207 nm. l-Citrulline was well separated using Gemini C-18 column and the identity of peak was confirmed by mass spectral analysis. These results were further confirmed by derivatisation method. The developed method was used for the quantification of l-citrulline in three varieties of watermelons such as Petite treat, Jamboree and Yellow crimson. Thus, the method is simple and involves direct analysis of aqueous extracts and can be used for rapid routine analysis of commercial samples. In addition, the role of watermelon extracts on smooth muscle relaxation markers was studied. Petite treat and Jamboree extracts induced production of nitric oxide (NO) significantly at 25 ppm. This was also confirmed by measuring the change in the intracellular calcium. Additionally, extracts of petite treat rind and juice have shown inhibition of phosphodiesterase-5A (PDE-5A) in human uterine smooth muscle cells (UtSMC). The induction of NO and the inhibition of PDE-5A seems to suggest the potency of these extracts to induce relaxation of smooth muscle. It is possible that watermelon extracts may have the potential to induce PDE-5A mediated smooth muscle relaxation. However, further studies are critical before any conclusion of smooth muscle relaxation effect.     

Substrate specificity of a recombinant d-lyxose isomerase from Providencia stuartii for monosaccharides

The specific activity and catalytic efficiency (k_cat/K_m) of the recombinant putative protein from Providencia stuartii was the highest for d-lyxose among the aldose substrates, indicating that it is a d-lyxose isomerase. Gel filtration analysis suggested that the native enzyme is a dimer with a molecular mass of 44 kDa. The maximal activity for d-lyxose isomerization was observed at pH 7.5 and 45 °C in the presence of 1 mM Mn²⁺. The enzyme exhibited high isomerization activity for aldose substrates with the C2 and C3 hydroxyl groups in the left-hand configuration, such as d-lyxose, d-mannose, l-ribose, d-talose, and l-allose (listed in decreasing order of activity). The enzyme exhibited the highest activity for d-xylulose among all pentoses and hexoses. Thus, d-lyxose was produced at 288 g/l from 500 g/l d-xylulose by d-lyxose isomerase at pH 7.5 and 45 °C for 2 h, with a conversion yield of 58 % and a volumetric productivity of 144 g l^− 1 h^− 1. The observed k_cat/K_m (920 mM^− 1 s^− 1) of P. stuartiid-lyxose isomerase for d-xylulose is higher than any of the k_cat/K_m values previously reported for sugar and sugar phosphate isomerases with monosaccharide substrates. These results suggest that the enzyme will be useful as an industrial producer of d-lyxose.     

Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine

The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain.     

Antioxidant activity of Lactobacillus plantarum strains isolated from traditional Chinese fermented foods

Eleven Lactobacillus plantarum strains isolated from traditional Chinese fermented foods were investigated for their in vitro scavenging activity against hydroxyl and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, and their resistance to hydrogen peroxide. L. plantarum C88 at a dose of 10(10) CFU/ml showed the highest hydroxyl radical and DPPH scavenging activities, with inhibition rates of 44.31% and 53.05%, respectively. Resistance of intact cells to hydrogen peroxide was also found in all strains. L. plantarum C88 was the most resistant strain against hydrogen peroxide. When L. plantarum C88 was administered to senescent mice suffering oxidative stress induced by d-galactose, the serum superoxide dismutase activity, the glutathione peroxidase activity and the total antioxidant capacity in liver increased significantly, while the level of malondialdehyde in liver decreased significantly. L. plantarum C88 isolated from traditional Chinese fermented dairy tofu could be considered as a potential antioxidant to be applied in functional foods.     

Evaluation of l-ascorbic acid oxidation on SH concentration in soy-wheat composite dough during resting period

The effect of l-ascorbic acid (l-AA) on free sulfhydryl concentration (SH) was evaluated in soy-wheat composite dough from 100-500 (g/kg) soy flour substitution for wheat flour. Raw soy flour (RSF) and physically modified soy flours (PMSF1 and PMSF2) were used for the preparation of the composite dough with wheat flour. The two physically modified soy flours were prepared by steam flushing (PMSF2) and water boiling (PMSF1) of raw soy beans before flour preparation. Using a timer, dough blends were manually mixed (at approximately 60 rpm) to dough development time after which, dough was sampled for the estimation of free SH groups. l-AA (0.05% w/w) was mixed with the dough after dough development and the dough was sampled after 1 h of resting the dough. The results showed that l-AA (0.05% w/w) acted as a reducing agent by increasing SH levels in all soy-wheat dough blends (P<0.05). After 1 h of resting, soy-wheat composite dough without l-AA had lower concentrations of SH than that with l-AA. A positive correlation was shown between soy flour concentrations and SH concentration before and after dough resting. A negative correlation existed between l-AA consumption and SH concentration for RSF-wheat, PMSF1-wheat and PMSF2-wheat doughs. The results indicated that soy flour weakened wheat flour dough by increasing SH concentration and that l-AA could have a synergistic effect on the reduction of gluten proteins and thus weakening the dough.