Structure characterization and antitumor activity of an α β-glucan polysaccharide from Auricularia polytricha

A fraction of a 0.05 M NaOH solution-soluble polysaccharide, termed AAFRC, was obtained from Auricularia polytricha by protease-assisted aqueous extraction followed by precipitation with ethanol and purification with gel filtration chromatography. Analysis by high performance liquid chromatography (HPLC) coupled to a TSK-G5000PW_XL column and gel filtration chromatography on Sephacryl S-400HR indicated that AAFRC is homogenous with an average molecular weight (Mw) of about 1.20 × 10⁶ Da. The structure of AAFRC was revealed by chemical methods including Fourier transform infrared (FT-IR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Results indicated that AAFRC is a glucan consisting 1,3-β-glucan, 1,4-α-glucan, and 1,3-α-glucan backbone with a single 1→)-α-d-glucopyranosyl side-branching unit on every six residues, on average, along the main chain. The growth of transplantable Sarcoma-180 (S₁₈₀) in mice was also significantly inhibited by AAFRC compared with the model controls (P < 0.01), with the inhibitory rate being 43.61%.     

Molecular cloning and characterization of a thermostable α-amylase exhibiting an unusually high activity

An α-amylase gene was cloned from the thermophilic bacterium Bacillus subtilis isolated from Indonesian oil palm shell waste. The gene expressed an extracellular enzyme. Optimal hydrolysis conditions for the enzyme were 70°C and pH 6.0. The specific activity of the enzyme was 16.0 kU per mg of protein, which was higher than for other thermostable amylases. Hydrolytic products of the enzyme using starch and glycogen were mainly maltohexaose and maltopentaose. The enzyme had a K _m value of 0.099 mg/mL for amylopectin, more than 10 times lower than for amylose. The catalytic efficiency of the enzyme using amylopectin was 39,200 mL/mg·s and was 3,270 mL/mg·s using amylose. The enzyme liquefied corn starch at pH 5.0, which was successfully converted to glucose using commercial glucoamylase and pullulanase without pH adjustment. The enzyme has advantages for industrial applications.     

Antioxidant properties of resveratrol: A structure–activity insight

Resveratrol, a natural product, is known to affect a broad range of intracellular mediators. In the present study, we clarified the antioxidant activity of resveratrol by employing various in vitro antioxidant assays such as DPPH?, ABTS^?+, DMPD^?+, O₂^?? and H₂O₂ scavenging activities, total antioxidant activity, reducing abilities, and Fe²⁺ chelating activities. Resveratrol inhibited 89.1% of the lipid peroxidation of linoleic acid emulsion at 30 µg/mL concentration. On the other hand, BHA, BHT, α-tocopherol, and trolox exhibited inhibitions of 83.3, 82.1, 68.1, and 81.3% against peroxidation of linoleic acid emulsion at the same concentration, respectively. In addition, resveratrol had effective DPPH?, ABTS^?+, DMPD^?+, O₂^?? and H₂O₂ scavenging activities, reducing power, and Fe²⁺ chelating activities. The present study found that resveratrol had effective in vitro antioxidant and radical scavenging activity. It can be used in pharmacological and food industry due to its antioxidant properties.Industrial relevanceAntioxidants are often added to foods to prevent the radical chain reactions of oxidation and they act by inhibiting the initiation and propagation step leading to the termination of the reaction and delay the oxidation process. At the present time, the most commonly used antioxidants are BHA, BHT, propylgallate and tert-butyl hydroquinone. Besides that BHA and BHT are restricted by legislative rules because of doubts over their toxic and carcinogenic effects. Therefore, there is a growing interest on natural and safer antioxidants in food applications, and a growing trend in consumer preferences for natural antioxidants, all of which has given more impetus to explore natural sources of antioxidants. A variety of foods and beverages of vegetable origin contain several nonflavonoid classes of phenolic compounds synthesized by plants. Among them, resveratrol has been identified as the major active compound of stilbene phytoalexins and is presumed to be beneficial for human health. Resveratrol is naturally occurring in the fruits and leaves of edible plants, peanuts, mulberries, grapes and red wine. Resveratrol is currently in the limelight all over the world due to their beneficial effects on the human body. Resveratrol can be used for minimizing or preventing lipid oxidation in pharmaceutical products, retarding the formation of toxic oxidation products, maintaining nutritional quality and prolonging the shelf life of food products and pharmaceuticals instead of BHA and BHT and other antioxidant compounds because of their safer usage.     

L-theanine—a unique amino acid of green tea and its relaxation effect in humans

Since ancient times, it has been said that drinking green tea brings relaxation. The substance that is responsible for a sense of relaxation, is theanine. Theanine is a unique amino acid found almost solely in tea plants and the main component responsible for the exotic taste of ‘green’ tea. It was found that L-theanine administered intraperitoneally to rats reached the brain within 30 min without any metabolic change. Theanine also acts as a neurotransmitter in the brain and decreased blood pressure significantly in hypertensive rats. In general, animals always generate very weak electric pulses on the surface of the brain, called brain waves. Brain waves are classified into four types, namely α,β,δ and θ-waves, based on mental conditions. Generation of α-waves is considered to be an index of relaxation. In human volunteers, α-waves were generated on the occipital and parietal regions of the brain surface within 40 min after the oral administration of theanine (50–200 mg), signifying relaxation without causing drowsiness. With the successful industrial production of L-theanine, we are now able to supply Suntheanine™ (trade name of L-theanine) which offers a tremendous opportunity for designing foods and medical foods targeting relaxation and the reduction of stress. Taiyo Kagaku Co., Ltd, Japan won the 1998 ‘Food Ingredient Research Award’ for development of Suntheanine™ at Food Ingredients in Europe (Frankfurt). The judges felt it was a particularly well-documented and fascinating piece of research.     

A large-scale isolation of native β-lactoglobulin: characterization of physicochemical properties and comparison with other methods

A novel method for β-lactoglobulin (β-lg) isolation from whey was investigated. The method comprised a peptic treatment of whey and membrane filtration under gentle conditions for isolation of native β-lg. The method was applied to 10,000 L batches of processed whey and the product quality obtained was compared with pilot-scale results of three other procedures: 3% trichloroacetic acid (TCA) precipitation, a salting-out procedure and a selective thermal precipitation. The calculated yields of native β-lg were 67.3, 44.9, 46.7 and 49.6% of the β-lg present in whey using the four methods, respectively. The composition of the different β-lg preparations was found to be comparable, however, analysis of purity revealed slight differences as indicated by electrophoresis and fast protein liquid chromatography (FPLC). The isolated β-lg retained a high degree of purity and native properties. The new method for isolation of native β-lg was found to be reproducible, selective and robust as processing of three 10,000 L batches did not indicate any technical problems or variations in the purity of the isolated β-lg.     

Antioxidant activity of yoghurt peptides: Part 2 – Characterisation of peptide fractions

The aim of the present study was to elucidate previous findings showing that peptide fractions isolated from yoghurt had antioxidant effects. Therefore, peptides and free amino acids released during fermentation of milk were characterised. Yoghurt samples were stripped from sugars and lactic acid and subsequently fractionated by ultra filtration using membranes with cut off sizes of 30, 10 and 3 kDa. The peptides in these fractions were identified by LC–MS/MS. The identified peptides comprised a few N-terminal fragments of αs₁-, αs₂-, and κ-casein, and several fragments from β-casein. Almost all the peptides identified contained at least one proline residue. Some of the identified peptides included the hydrophobic amino acid residues Val or Leu at the N-terminus and Pro, His or Tyr in the amino acid sequence, which is characteristic of antioxidant peptides. In addition, the yoghurt contained a considerable amount of free amino acids such as His, Tyr, Thr and Lys, which have been reported to have antioxidant properties. Thus, our findings confirm that the antioxidant effects of the peptide fractions from yoghurt are due to the presence of certain peptides and free amino acids with recognised antioxidant activity in these fractions.     

Kinetics and equilibria of tea infusion: Part 8—the effects of salts and of pH on the rate of extraction of theaflavins from black tea leaf

The rates of extraction of theaflavins from sieved Kapchorua Pekoe Fannings (600–710 μm) and from a Ceylon Broken Orange Pekoe Fine Leaf Blend have been measured at 80°C with a set of aqueous salt and buffer solutions spanning the pH range 3·1 to 8·1. There was little effect on adding 1:1 electrolytes but considerable decreases in rate occurred in the presence of calcium chloride. The rate constants in acid buffers were the same as in a normal unbuffered infusion but in alkaline buffers of pH 8 they were over 50% larger. This phenomenon was quantitatively accounted for by the partial dissociation of theaflavins at higher pH and the greater diffusion coefficient of the resulting ionic salt. The relevance of these findings to tea brewing and to industrial theaflavins extraction is discussed.     

Antioxidant activity and components of Salvia plebeia R.Br. — a Chinese herb

The antioxidant properties of the extracts from Salvia plebeia R.Br. which was screened out of over 700 species of Chinese herbs, were tested in lard at 110°C using the Oxidative Stability Instrument. The ethyl acetate extract of the herb was re-extracted by solvents (with increasing polarity) into petroleum ether-, diethyl ether-, acetone-and-ethanol-soluble fractions. The fractions obtained were then separated according to their acidic properties. From the most active sub-fractions, namely the acidic sub-fraction of the petroleum ether-soluble fraction and the weakly acidic sub-fraction of the diethyl ether-soluble fraction, three antioxidant components were isolated and identified as royleanonic acid, hispidulin and eupatorin. The royleanonic acid was a novel compound and eupatorin was isolated for the first time from the herb. Though royleanonic acid and hispidulin prolonged the induction period significantly, their antioxidant activities were much weaker than the crude extracts, implying that synergistic effects might be responsible for the high activity of the crude extracts.     

Quantification of folpet and phthalimide in tea and herbal infusions by LC– high-resolution MS and GC–MS/MS

Two methods based on a modified QuEChERS sample preparation and either LC coupled to atmospheric pressure ionisation and high-resolution MS or GC coupled to electron ionisation and tripled quadrupole MS have been assessed for the quantification of folpet and phthalimide in tea and other dry herbal infusions. Both methods have been fully validated in green tea and further checked in black tea, verbena and rooibos, and they performed according to the SANTE/11813/2017 criteria at the target LOQ concentration level (50 µg/kg). These methods allow the accurate quantification of folpet in the selected matrices according to the new EU residue definition, which includes phthalimide. Phthalimide is the main metabolite and degradation product of folpet, although according to recent studies, it could be generated from different sources than folpet breakdown, such as food processing or analysis by GC.     

Antioxidant activity of tomato lipophilic extracts and interactions between carotenoids and α-tocopherol in synthetic mixtures

In the present study we assayed the antioxidant activity of lipophilic extracts obtained from different tomato varieties. The results showed that cherry tomatoes, characterized by a high carotenoid content, had the highest antioxidant activity. A quantitative analysis of lycopene, β-carotene, lutein and α-tocopherol was also performed and the correlation between the antioxidant content and the antioxidant activity was estimated. The highest correlation coefficient was found for lycopene (R² = 0.9236, P ≤ 0.001). The analysis of two-component mixtures containing α-tocopherol and carotenoids showed that significant synergism occurred for all the combinations which contained α-tocopherol and β-carotene mixed together. The highest synergistic effects were detected for α-tocopherol-lycopene mixtures, which were the most efficient combinations tested in the present study. The analysis of the carotenoid combinations indicated that synergism occurred for lycopene-β-carotene, lycopene-lutein and lutein-β-carotene mixtures. The analysis of four-component mixtures did not show statistically significant synergistic effects.     

Steam explosion improves extractability,antioxidant activity and α-glucosidase inhibitory activity of the constituents of Java tea (Clerodendranthus spicatus)

In this work, the steam explosion (SE) technology was exploited to extract valuable phytochemicals from Java tea (Clerodendranthus spicatus). Modification effects of SE on apparent morphology and physicochemical properties of Java tea were investigated to confirm that SE destroyed the cell wall structure and enhanced pore space, which facilitated the dissolution of ingredients. After SE pretreatment, the contents of phenolics, triterpenes, and sterols of Java tea extracts were maximally increased by 40.68%, 25.69%, and 40.11% in the leaves group, and increased by 147.09%, 204.50%, and 115.24% in the stems group, respectively. The antioxidant and α-glucosidase inhibition capacity of extracts were also obviously enhanced. Multivariate analysis indicated that phenolics and triterpenes were the main active substances in the extracts, and the leaves treated at 1.0 MPa and stems treated at 2.4 MPa showed higher comprehensive score. This study provided a promising method for Java tea pretreatment for improving extractability and bioactivity.