Health-promoting activities of ultra-filtered okara protein hydrolysates released by in vitro gastrointestinal digestion: identification of active peptide from soybean lipoxygenase

Okara, a major by-product of the soymilk industry, which is rich in proteins, could possibly release under physiological conditions potential bioactive peptides. Thus, an okara protein isolate was digested sequentially with pepsin and pancreatin for c.a. 4 h. On the basis of its relatively high degree of hydrolysis and antioxidant activities (power reduction and radical scavenging activity), the okara protein hydrolysates at the end of the in vitro digestion were fractionated by ultra-filtration and the obtained ultra-filtered fractions were further tested for angiotensin-converting enzyme inhibition and multifunctional antioxidant activities. In the <1 kDa molecular weight cutoff ultra-fraction the amino acid sequence, TIIPLPV, of a peptide from soybean lipoxygenase-1 with a calculated mass 751.48 Da was identified using LC-ESI-MS/MS techniques. The hydrophobic amino acids present in this peptide, particularly Val at terminal position, could likely be associated with the relatively high health-promoting attributes tested. This study evidenced that the consumption of okara protein may exert health benefits on the basis of the bioavailability and bioactivity of the identified peptide.     

Antioxidant activity of seed polyphenols in fifteen wild Lathyrus species from South Spain

Antioxidant activity of seed phenolics was studied in the following Lathyrus species: Lathyrus hirsutus, Lathyrus filiformis, Lathyrus sativus, Lathyrus cicera, Lathyrus angulatus, Lathyrus sphaericus, Lathyrus annuus, Lathyrus clymenum, Lathyrus pratensis, Lathyrus ochrus, Lathyrus aphaca, Lathyrus latifolius, Lathyrus setifolius, Lathyrus tingitanus and Lathyrus amphicarpos. Phenolic contents ranged from 3.8 mg/g meal in L. setifolius to 29.2 mg/g meal in L. sphaericus. In general, non-cultivated Lathyrus species contained higher phenolic contents than cultivated ones. A negative correlation between seed size and phenolic contents was observed and was related to the higher proportion of hulls in the smaller seeds. L. annuus possessed phenolics with highest specific antioxidant activity. These phenolics were more than two times more antioxidant than equivalent amounts of phenolics extracted from commercial chickpea, lupin or soy. On the other hand, L. aphaca possessed the highest antioxidant activity per mg of flour extract. This antioxidant activity was twice that observed in same amounts of extracted flours from commercial chickpea, lupin or soy. Results show that studied Lathyrus species are rich in phenolic compounds with higher antioxidant activity than phenolics of widely consumed legumes such as soy, chickpea or lupin. In conclusion, Lathyrus may represent an interesting source of phenolic compounds with high antioxidant activity that may be useful as natural antioxidants and contribute to revalorize the cultivation of these legumes.     

Nutritional characteristics of seed proteins in 15 Lathyrus species (fabaceae) from Southern Spain

The nutritional characteristics of seed proteins of 15 Spanish Lathyrus species have been analyzed. Protein contents in studied Lathyrus ranged from 17.7% in Lathyrus sativus to 25.6% in L. tingitanus with a 22.4% average protein content in studied Lathyrus. Among essential amino acids the most abundant were Leu, Lys, Phe, Thr and Val. Also, all species contained Lys above FAO recommendations. On the contrary, all species were limiting in Trp ranging from 0.5% to 0.8% content and were also limiting in sulphur containing amino acids, Met and Cys. In vitro protein digestibility (IVPD) in studied Lathyrus ranged from 76.0% in L. pratensis to 86.5% in L. annuus, with an average IVPD of 80.5%. Nutritional parameters, such as protein efficiency ratio, corresponded to high quality proteins. Protein digestibility corrected amino acid score was similar to the observed in other legumes such as lentil or peanut. An analysis of similarity based on the profile of seed protein amino acid composition showed, in the cluster generated, correlation with the taxonomic classification of these species according to morphological characters. Results confirm the interest of studying wild populations of cultivated and non-cultivated Lathyrus species as source of seeds with good nutritional characteristics.     

Cell proliferation inhibitory and apoptosis-inducing properties of anacardic acid and lunasin in human breast cancer MDA-MB-231 cells

The combination of two or more chemopreventive agents is currently being used to achieve greater inhibitory effects on breast cancer cells. Anacardic acid and lunasin are two plant-derived compounds that have been associated with anti-carcinogenic properties. These compounds show inhibitory effects on cell proliferation and inducing properties of apoptosis in human breast cancer MDA-MB-231 cell line. Both lunasin and anacardic acid exert their effects through the modulation of expression of several genes involved in cell cycle, apoptosis and signal transduction. Their combination arrests the cell cycle in S-phase and induces apoptosis at higher levels than that observed when each compound is used individually. This combination also promotes the inhibition of ERBB2, AKT1, JUN and RAF1 signalling gene expression, whose up-regulation has been reported as responsible for breast cancer cells growth and resistance to apoptosis. Our results introduce these two compounds as a promising strategy to prevent/treat breast cancer.     

Affinity purification and characterisation of chelating peptides from chickpea protein hydrolysates

A chickpea protein hydrolysate produced with pepsin and pancreatin was used for the affinity purification of chickpea chelating peptides. Three chelating peptide fractions were obtained after affinity chromatography with immobilised copper. These peptide fractions showed a higher chelating activity and histidine contents than the original protein hydrolysate. Chelating activity was positively correlated with the histidine content of the purified fractions. Different subfractions were also obtained after gel filtration chromatography from the affinity purified peptide fractions. Some of these subfractions showed a higher chelating activity and histidine contents than the original fractions. These results suggest that a combination of high His contents, around 20–30%, and small peptide size provide the best chelating activities. Thus sequential purification with affinity and gel filtration chromatography is a useful procedure for the purification of chickpea peptides with high chelating activity. These results show that a range of chelating peptides are generated during digestion of the chickpea proteins that, after metal chelation, may prevent the generation of reactive oxygen species (ROS) and favour metal absorption.     

Degradation of tannic acid by cell-free extracts of Lactobacillus plantarum

The ability of Lactobacillus plantarum CECT 748^T to degrade hydrolysable tannins was evaluated. Three commercial tannic acids were incubated in presence of cell-free extracts containing soluble proteins from L. plantarum. By HPLC analyses, almost a complete tannic acid degradation was observed in the three samples assayed. By using HPLC-DAD/ESI-MS, we partially determined the composition of tannic acid from Quercus infectoria galls. This tannic acid is a gallotannin mainly composed of monomers to tetramers of gallic acid. We studied the mechanism of its degradation by L. plantarum. The results obtained in this work indicated that L. plantarum degrades gallotannins by depolymerisation of high molecular weight tannins and a reduction of low molecular weight tannins. Gallic acid and pyrogallol were detected as final metabolic intermediates. Due to the potential health beneficial effects, the ability to degrade tannic acid is an interesting property in this food lactic acid bacteria.     

Efficacy of recA gene sequence analysis in the identification and discrimination of Lactobacillus hilgardii strains isolated from stuck wine fermentations

Conventional phenotypic methods sometimes lead to misidentification of some heterofermentative wine lactobacilli such as Lactobacillus hilgardii, Lactobacillus buchneri, and Lactobacillus brevis. We establish the specificity of 16S rDNA sequencing in the differentiation of these species and in the rejection of the Lactobacillus vermiforme species name. Moreover, we succeeded in differentiating these heterofermentative species by means of recA gene sequence comparison. Short homologous regions were amplified by PCR with degenerate consensus primers, sequenced, and 280 bp were analysed and considered for the inference of phylogenetic trees. The phylogram obtained was coherent and clearly separated the three species. The recA gene sequence was a reliable and useful method that allowed a good discrimination among closely related species. The validity of the recA gene sequence, restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified 16S-23S rDNA intergenic spacer region (ISR), and random amplified polymorphic DNA (RAPD) to study the L. hilgardii intraspecies heterogeneity was tested in five strains isolated from stuck wine fermentations at the same winery in the same vintage. The results indicated that L. hilgardii is a heterogeneous species. Since L. hilgardii is a malolactic species that can influence the final quality of the wine, the presence of oenological relevant genes, such as those involved in ethyl carbamate or biogenic amine production, was investigated.     

High Hydrostatic Pressure and Temperature Applied to Preserve the Antioxidant Compounds of Mango Pulp (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.)

High hydrostatic pressure (HHP) processes combined with moderate heating can be used to preserve foods while maintaining general quality. The effect of these conditions on the total phenolic (TP), vitamin C (L-ascorbic acid (AA)), carotenoids, and antioxidant activity (AOA) of mango purees was evaluated. Purees were processed at 400–550 MPa/34 and 59 °C at different holding times. Unprocessed puree had TP of 26.6 mg gallic acid/100 g, 21.1 mg L-ascorbic acid/100 g, AOA of 885 μmol trolox equivalents/100 g, and total carotenoids of 6.0 mg β-carotene/100 g. HHP treatments increased the phenolic concentrations up to 34% (550 MPa/59 °C/2 and 4 min) compared with the initial content, probably due to improvement of their extraction. AA content was reduced significantly (10–45%) after all HHP processes performed at 59 °C, while at 34 °C, they were diminished only after 8 and 16 min of treatment (13–26%). At 34 °C and lower times, AA concentration increased in average 18%. Total carotenoid retention in HHP-treated samples varies from 77 to 98%, being the higher the temperature the lower the retention observed. The concentration of most individual carotenoids remains unchanged, but violaxanthin content was reduced (21–26%) and 9-cis-violaxanthin was increased by about 10%. The AOA was also increased (up to 39%) at some processing conditions. A linear correlation between the TP and AOA was obtained. HHP at 550 MPa combined with moderate temperature (34 °C) at processing times up to 8 min is recommended for the maximum retention of the antioxidant compounds of mango puree.     

Production,partial purification and properties of β‐mannanases obtained by solid substrate fermentation of spent soluble coffee wastes and copra paste using Aspergillus oryzae and Aspergillus niger

In order to achieve a higher added value of two galactomannan‐containing wastes, copra paste and spent coffee from the soluble coffee industry (SCW), solid substrate fermentation (SSF) was used. Filamentous fungi Aspergillus oryzae and A niger were used to evaluate the feasibility of producing β‐mannanase by SSF. A 2³ factorial design was used to select the best interaction among the two fungi, the two substrates and two fermentation times. The treatment ‘A niger–copra–2.5 days’ produced a significantly higher (p < 0.05) β‐mannanase activity, having five different isoforms of the enzyme, one of which was partially purified to a specific activity of 764 U mg⁻¹ (U = nmol of mannose released per second from a galactomannan substrate). Copra paste had a higher mannose/galactose ratio (14:1) than SCW (6:1), and low oil content, which led to higher β‐mannanase production from SSF. A β‐mannanase from SSF of copra produced by A oryzae was highly purified using acetone precipitation and cation exchange and size exclusion chromatographies. This enzyme had an MW of 110 kDa, a pI between 3.5 and 4.5 and a specific activity of 1760 U mg⁻¹; purification achieved was 90.7 times. The temperature and pH for optimal activity were 40 °C and 6.0 respectively. The optimal temperature was lower and the optimal pH higher than others previously reported (produced by submerged fermentation), which could be important for viscosity reduction of concentrated coffee extract in instant coffee manufacture. Copra is an interesting alternative for β‐mannanase production, since it is readily available in Mexico; moreover, the residue after SSF has a reduced galactomannan content and may be used for monogastric animal feed. © 2000 Society of Chemical Industry