Effect of thermal and high pressure processes on structural and health-related properties of carrots (Daucus carota)

In the present study, the effect of thermal and equivalent high pressure processes on structural (texture and microstructure) and health-related (total β-carotene concentration, β-carotene isomerisation and β-carotene bio-accessibility) properties of carrots was investigated. Both a mild and strong pasteurisation process and a sterilisation process were considered.     

Precipitation of β-carotene microparticles from SEDS technique using supercritical CO2

This work investigates the application of the Solution-Enhanced Dispersion by supercritical fluids technique for the precipitation of β-carotene. The effect of pressure (8.0–12.0 MPa), temperature (293–313 K) anti-solvent flow rate (20–40 mL/min), solution flow rate (1–4 mL/min) and concentration of β-carotene in the dichloromethane solution (4 and 8 mg/mL) on the precipitation yield, particle morphology and particle size and size distribution was examined. Precipitated powders presented mean particle size varying from 3.2 μm to 96.8 μm with morphology of β-carotene microparticles changing from plate-like to leaf-like particles. The statistical analysis of the experimental results revealed that pressure, organic solution concentration and CO₂ flow rate had a significant effect on particle size. The precipitation yield was observed to be within the range of 71–94% and was statistically influenced by system temperature and pressure, and anti-solvent flow rate.     

Supercritical fluid extraction of β-carotene from crude palm oil using CO2

The aim of this study was to ascertain the influence of pressure, temperature and time on the supercritical fluid extraction of β-carotene from the crude palm oil. The operating conditions were shown as follows: pressures of 75, 125 and 175 bar, temperatures of 80, 100 and 120 °C and extraction time of 1, 3 and 5 h. The extracts were analyzed using UV spectroscopy at a wavelength of 450 nm. Then the experimental data was compared with the data obtained using a statistical method. The results from the model showed a good agreement with the experimental data. The results (obtained from the statistical model) demonstrate that a pressure of 140 bar, temperature of 102 °C and extraction time of 3.14 h are required to obtain optimum yield of β-carotene (1.028 × 10⁻²%) from the extraction process, however the maximum yield of the β-carotene (1.741 × 10⁻²%) was experimentally obtained at a pressure of 75 bar, temperature of 120 °C and extraction time of 1 h.     

Heat-induced aggregation of β-lactoglobulin A and B with α-lactalbumin

β-Lactoglobulin A and β-lactoglobulin B were heated at 75°C in the absence and presence of α-lactalbumin, and the aggregation products were characterized by size exclusion chromatography in combination with multi-angle laser light scattering and electrophoretic techniques. α-Lactalbumin did not form aggregates when heated alone, but in admixture with β-lactoglobulin it was incorporated into both the disulphide-bonded and the hydrophobically associated aggregates as well as forming α-lactalbumin dimers and other oligomers. The presence of α-lactalbumin diminished the proportion of smaller aggregates and increased the number of very large aggregates within both variant protein mixtures. In the presence of α-lactalbumin, β-lactoglobulin A was converted into a series of disulphide-bonded and the hydrophobically associated aggregates more slowly, but with a greater proportion of hydrophobically associated aggregates, than β-lactoglobulin B. These patterns are similar to that when β-lactoglobulin A or B are heated on their own. These and other results indicate that the mechanism of aggregation of α-lactalbumin/β-lactoglobulin mixtures is governed by β-lactoglobulin.     

Release and absorption of carotenes from processed carrots (Daucus carota) using in vitro digestion coupled with a Caco-2 cell trans-well culture model

The release and absorption (bioavailability) of carotenoids is a prerequisite for their nutritional impact. This can be strongly affected by the processing conditions used to prepare the food matrix that contains them. To determine the effect of processing on carotenoid bioavailability, homogenized, raw, blanched and cooked carrots were exposed to an in vitro gastric and intestinal digestion model. Final digest samples were placed onto a Caco-2 cell trans-well monolayer culture to mimic intestinal absorption. The results show that the cooked carrot puree consisting of primarily single plant cell particles had the highest release of carotenes, followed by blanched consisting primarily of plant cell clusters and raw carrot puree consisting of larger plant cell clusters. Absorption through the Caco-2 cell layer was the highest from the digesta of cooked carrot puree followed by the digesta of blanched carrot puree. This study demonstrates that thermal processing and/or mechanical homogenization to disrupt plant cell wall matrix enhances the in vitro bioavailability of carotenes from carrots.     

DETERMINATION OF α- AND β-AMYLASE IN MALT

The Analysis Committee of the European Brewery Convention carried out a collaborative trial on malts using the specific analysis methods for α- and β-amylase activities based on dyed substrates supplied by MegaZyme (Aust.) Pty. Ltd. The repeatability and reproducibility values for the methods were judged to be unsatisfactory and consequently the methods were not recommended for Analytica-EBC.     

β-carotene content in sweet potato varieties from China and the effect of preparation on β-carotene retention in the Yanshu No. 5

To compare the β-carotene contents in different Chinese sweet potato (SP) varieties and to choose a variety of SP rich in β-carotene for the study of the effect of processing methods on β-carotene retention, β-carotene in thirteen varieties of sweet potato from China was measured by HPLC. The results showed that β-carotene contents were significantly correlated with SP flesh colours, with the orange-red fleshed SP varieties being higher in β-carotene.β-carotene contents in SP were affected by many factors, and this was demonstrated using the variety of Yanshu No.5, showing that the β-carotene contents in SP grown in different farming sites in the same area ranged from 53.2 to 84.3 mg kg^− 1 fresh weight. Moreover, β-carotene distributes unevenly in one SP root, with highest concentrations in the core. The β-carotene content was positively related to the root size.Five processing methods including boiling, steaming, microwave cooking, frying, and post steam-drying were simulated in the study to check their effects on the true retention of β-carotene in SP. Compared to boiling, steaming resulted in much more loss of β-carotene and microwave cooking resulted in the biggest loss of β-carotene among the five processing methods.

Industrial relevance

Orange-fleshed sweet potato can be prepared for sale and consumption, using methods that protect the β-carotene content. This can aid in promoting sweet potato as a staple food as well as a snack food for supplying vitamin A for both rural and urban populations. Prepared orange-fleshed sweet potato can contribute to alleviating vitamin A deficiency in China as well as other low-income countries.     

Effects of Processing on the Reduction of β-ODAP (β-N-Oxalyl-L-2,3-diaminopropionic acid) and Anti-Nutrients of Khesari Dhal,Lathyrus sativus

Effects of different processing techniques on the neurotoxin, β-ODAP (β- N -oxalyl-L-2,3-diaminopropionic acid), and the anti-nutritional compounds (phytate, polyphenols, trypsin and amylase inhibitors, and lectins) within four lines of Lathyrus sativus (high-, medium- and low-ODAP, and so-called ODAP-free) were investigated. Soaking of seeds in various media reduced the contents of these compounds to a varying and significant extent; losses were higher in freshly boiled water, alkaline and tamarind solutions than after soaking in drinking water. The highest losses in boiled water (65–70%) were observed for β-ODAP, followed by trypsin inhibitors (42–48%) and polyphenols (30–37%). Ordinary cooking and pressure cooking of pre-soaked seeds were found to be most effective in reducing the levels of all the natural toxicants examined, whilst fermentation and germination were more effective in destroying both of the enzyme inhibitors (amylase inhibitors by 69–71%; trypsin inhibitors by 65–66%) than either phytates or polyphenols. Lectins were not affected by most of these processes except by pressure cooking and fermentation. Dehusking of pre-soaked seeds significantly reduced β-ODAP levels, but this reduction was lower for the anti-nutrients. These findings and the high water solubility suggest that a simple and effective means of detoxifying Lathyrus by removing this neurotoxic amino acid may be practicable.     

Fractionation and identification of ACE-inhibitory peptides from α-lactalbumin and β-casein produced by thermolysin-catalysed hydrolysis

The thermolysin catalysed hydrolysates of α-lactalbumin and β-casein were fractionated by size-exclusion chromatography (SEC) and reversed-phase high performance liquid chromatography (RP-HPLC) in order to identify the peptides responsible for the high ACE-inhibitory activity of these hydrolysates. The SEC fractionation separated many co-eluting peptides into different fractions allowing individual peptides to be isolated in one or two subsequent semi-preparative RP-HPLC fractionation steps. Five potent ACE-inhibitory peptides from α-lactalbumin were isolated. They all contained the C-terminal sequence -PEW, corresponding to amino acid residues 24–26 in α-lactalbumin, and had IC₅₀ values of 1–5 μm. From one SEC fraction of the β-casein hydrolysate two potent ACE-inhibitory peptides were isolated and identified as f58-76 and f59-76 of β-casein A2. They both contained IPP as the C-terminal sequence and had IC₅₀ values of 4 and 5 μm. From another SEC fraction a new but less ACE-inhibitory peptide from β-casein was identified (f192–196; LYQQP).     

ENZYMIC QUANTIFICATION OF (1→3) (1→4)-β-D-GLUCAN IN BARLEY AND MALT

A simple and quantitative method for the determination of (1→3) (1→4)-β-D-glucan in barley flour and malt is described. The method allows direct analysis of β-glucan in flour and malt slurries. Mixed-linkage β-glucan is specifically depolymerized with a highly purified (1→3) (1→4)-β-D-glucanase (lichenase), from Bacillus subtilis, to tri-, tetra- and higher degree of polymerization (d.p.) oligosaccharides. These oligosaccharides are then specifically and quantitatively hydrolysed to glucose using purified β-D-glucosidase. The glucose is then specifically determined using glucose oxidase/peroxidase reagent. Since barley flours contain only low levels of glucose, and maltosaccharides do not interfere with the assay, removal of low d.p. sugars is not necessary. Blank values are determined for each sample allowing the direct measurement of β-glucan in maltsamples.α-Amylasedoes not interfere with the assay. The method issuitable for the routineanalysis of β-glucan in barley samples derived from breeding programs; 50 samples can be analysed by a single operator in a day. Evaluation of the technique on different days has indicated a mean standard error of 0–1 for barley flour samples containing 3–8 and 4–6% (w/w) β-glucan content.     

Factors influencing the production of o/w emulsions stabilized by β-lactoglobulin–pectin membranes

A primary emulsion was prepared by homogenizing 10 wt% corn oil with 90 wt% aqueous β-lactoglobulin solution (0.5 wt% β-lg, pH 3 or 7) using a two-stage high-pressure valve homogenizer. This emulsion was mixed with aqueous pectin (citrus, 59% DE) stock solution (2 wt%, pH 3 or 7) and NaCl solution to yield secondary emulsions with 5 wt% corn oil, 0.225 wt% β-lactoglobulin, 0.2 wt% pectin and 0 or 100 mM NaCl. The final pH of the emulsions was then adjusted (3–8). Primary and secondary emulsions were ultrasonically treated (30 s, 20 kHz, 40% amplitude) to disrupt any flocculated droplets. Secondary emulsions were more stable than primary emulsions at intermediate pHs. Secondary emulsions prepared at pH 7 had smaller particle diameters (0.35 to 6 μm) than those prepared at pH 3 (0.42 to 18 μm) across the whole pH range studied, and also had smaller diameters than the primary emulsions (0.35 to 14 μm). Ultrasound treatment reduced the particle diameter of both primary and secondary emulsions and lowered the rate of creaming. The presence of NaCl screened the charges and thus the electrostatic interaction between biopolymer molecules and primary emulsion droplets. Secondary emulsions were more stable to the presence of 100 mM NaCl at low pHs (3–4) than primary emulsions. This study shows that stable emulsions can be prepared by engineering their interfacial membranes using the electrostatic interaction of natural biopolymers, especially at intermediate pHs where proteins normally fail to function.     

Emulsifying properties and structural characteristics of β‐conglycinin and dextran conjugates synthesised in a pressurised liquid system

The purpose of this study was to characterise the Maillard reaction in a pressurised liquid system containing β‐conglycinin and dextran. The rate of browning and the free amino content in this liquid system, with an initial pH of 7.2, were measured after reaction under atmospheric pressure and at 10 MPa. The secondary and tertiary structures of the conjugates were analysed by circular dichroism, showing that the increased pressure accelerated the reaction rate of the free amino groups and reduced the rate of browning. Significant structural changes occurred in the tertiary structure of the conjugates. Additionally, the emulsifying properties of the β‐conglycinin–dextran conjugates produced were studied. The emulsifying properties of the conjugates produced at 10 MPa were higher than those made at atmospheric pressure.     

Cloning and Sequence Analysis of the estA gene encoding enzyme for producing (R)-β-acetylmercaptoisobutyric acid from Pseudomonas aeruginosa 1001

The estA gene encoding the enzyme that catalyzes the production of (R)-β-acetylmercaptoisobutyric acid from (R,S)-ester from Pseudomonas aeruginosa 1001, was cloned in Escherichia coli and its nucleotide sequence was determined, revealing the presumed open reading frame encoding a polypeptide of 316 amino acid residues (948 nucleotides). The overall A+T and C+G compositions were 32.59% and 67.41%, respectively. The amino acid sequence of the estA gene product showed a significant similarity with that of the triacylglycerol lipase from Psychrobacter immobilis (38% identity), triacylglycerol lipase from Moraxella sp. (36% identity), and two forms of carboxyl esterases from Acinetobacter calcoaceticus (17% and 17% identities). The deduced amino acid sequences have a pentapeptide consensus sequence, G-X-S-X-G, having an active serine residue, and another active site, dipeptides H-G, located at 70–100 amino acids upstream of the G-X-S-X-G consensus sequence.     

Structural modification and characterization of rice starch treated by Thermus aquaticus 4-α-glucanotransferase

Rice starch was modified using Thermus aquaticus 4-α-glucanotransferase (TAαGTase) in this study. The changes in the molecular structure and the effect on the starch retrogradation by TAαGTase treatment were investigated on isolated rice starch. By treating TAαGTase, molecular weight profile of amylopectins shifted to higher elution time from 1.0 × 10⁸ to 2.4 × 10⁷ or 0.8 × 10⁷, depending on the level of enzyme dosage. Meanwhile, there were huge increases in the proportions of content corresponding to amylose size and even smaller molecules. On treating with TAαGTase, short branch chains (DP 1–8) increased, and longer branch chains (>DP 19) increased significantly as well, with a broader distribution up to DP 46 compared to the control rice starch. Amylose content decreased from 30.0 to 21.8–23.7%. This indicated that the amylose could be transferred to the amylopectin branch chain by the disproportionation of TAαGTase, resulting in lowering the amylose content and the formation of amylopectin with a broader branch-chain length distribution. TAαGTase modified rice starch showed that X-ray diffraction pattern of the B-type crystalline even before cold storage, and that a variety of cyclic glucans (DP 5–19) were produced by enzymatic reaction. In particular, the accelerated rate of starch retrogradation was clearly observed compared to the control due to an overall increase in the number of elongated long-branch chains, decrease in the amount of amylose–lipid complex, and the possible synergistic effects of these factors.     

Biotransformation of (R)-(+)- and (S)-(−)-limonene to α-terpineol by Penicillium digitatum— investigation of the culture conditions

The biotransformation of (R)-(+)- and (S)-(−)-limonene by Penicillium digitatum was investigated. One strain of P. digitatum was able to convert (R)-(+)-limonene to pure (R)-(+)-α-terpineol in 8 h with a yield of up to 93%. It was found that (R)-(+)-limonene was converted much better into α-terpineol than (S)-(−)-limonene, and that no significant chemical conversion of the substrate occurred in control flasks at pH 3.5. The culture conditions involved such as the type and concentration of co-solvent applied and the sequential addition of substrate were investigated, taking into account some findings on the physical behaviour of the system. The highest bioconversion yields were obtained when the substrate was applied as a diluted solution in EtOH.     

Measurement of (1 → 3),(1 → 4)-β-D-glucan in barley and oats: A streamlined enzymic procedure

A commercially available enzymic method for the quantitative measurement of (1 → 3),(1 → 4)-β-glucan has been simplified to allow analysis of up to 10 grain samples in 70 min or of 100–200 samples by a single operator in a day. These improvements have been achieved with no loss in accuracy or precision and with an increase in reliability. The glucose oxidase/peroxidase reagent has been significantly improved to ensure colour stability for periods of up to 1 h after development. Some problems experienced with the original method have been addressed and resolved, and further experiments to demonstrate the quantitative nature of the assay have been designed and performed.     

Interaction of β‐casein with (−)‐epigallocatechin‐3‐gallate assayed by fluorescence quenching: effect of thermal processing temperature

The interactions between the flavan‐3‐ol (?)‐epigallocatechin‐3‐gallate (EGCG) and bovine β‐casein in phosphate‐buffered saline (PBS) of pH 6.5 subjected to thermal processing at various temperatures (25–100 °C) were investigated using fluorescence quenching. The results indicated that different temperatures had different effects on the structural changes and EGCG‐binding ability of β‐casein. At temperatures below 60 °C, the β‐casein–EGCG interaction changed little (P > 0.05) with increasing temperature. At temperatures above 80 °C, native assemblies of β‐casein in solution dissociated into individual β‐casein molecules and unfolded, as demonstrated by a red shift of the maximum fluorescence emission wavelength (λ_max) of up to 8.8 nm. The highest quenching constant (K_q) and the number of binding sites (n) were 0.92 (±0.01) × 10¹³ m ^?1 s^?1 and 0.73 (±0.02) (100 °C), respectively. These results provide insight into the potential of interactions between β‐casein–EGCG that may modulate bioactivity or bioavailability to be altered during thermal process.     

Phenolic contents and antioxidant activities of bitter gourd (Momordica charantia L.) leaf, stem and fruit fraction extracts in vitro

Bitter gourd (Momordica charantia L.) has long been regarded as a food and medicinal plant. We investigated the antioxidant activity of the water extract of leaf, stem and fruit fractions by several in vitro systems of assay, namely DPPH radical-scavenging activity, hydroxyl radical-scavenging activity, β-carotene–linoleate bleaching assay, ferric reducing/antioxidant power (FRAP) assay and total antioxidant capacity. Total phenolic content was measured by Folin–Ciocalteu reagent. Identification of phenolic compounds was achieved using HPLC with the UV-diode array detection. The extracts of different fractions were found to have different levels of antioxidant activity in the systems tested. The leaf extract showed the highest value of antioxidant activity, based on DPPH radical-scavenging activity and ferric reducing power, while the green fruit extract showed the highest value of antioxidant activity, based on hydroxyl radical-scavenging activity, β-carotene–linoleate bleaching assay and total antioxidant capacity. The predominant phenolic compounds were gallic acid, followed by caffeic acid and catechin. The present study demonstrated that the water extract fractions of bitter gourd have different responses with different antioxidant methods. Total phenol content was shown to provide the highest association with FRAP assay in this present study (R² = 0.948).