Honey major protein characterization and its application to adulteration detection

The major proteins in honey have different molecular weights depending upon the honeybee species. To confirm the origin of major honey proteins, honey protein produced by Apis cerana or Apis mellifera were purified and analyzed by MALDI-TOF. Two major proteins were identified as a major royal jelly protein 1. Although two major proteins shared primary structure, they showed different molecular weights of 56 and 59 kDa, respectively. To discriminate the honeybee species producing honey using SDS–PAGE, artificial marker proteins, 56 and 59 kDa, were produced from Escherichia coli. Two artificial marker proteins were co-electrophoresed with honey samples and the difference in molecular weight was readily distinguished by SDS–PAGE. Therefore, the measurement of major proteins in honey is a useful method to discriminate the honey that produced from different honeybee species.     

Comparative study of the physicochemical and palynological characteristics of honey from Melipona subnitida and Apis mellifera

Twenty‐four samples of Apis mellifera honey and twenty‐four samples of Melipona subnitida (Jandaira) honey were collected in the northeast of Brazil. Moisture, hydroxymethylfurfural, free acidity, insoluble solids in water, diastase activity, ashes, electrical conductivity, proteins, lipids, total carbohydrates, energy and sugars were the parameters analysed. The efficiency of the qualitative tests (Fiehe's test, Lugol's reaction, Lund's reaction) was tested. Pollen types and the corresponding plant species were identified in all samples (3 in Apis and 1 in Melipona). Apis mellifera honey samples demonstrated parameters in accordance with the Brazilian Legislation, while the Melipona subnitida honey samples displayed moisture (24.80%) and diastase activity (null) in discordance with the established by the regulation for Apis mellifera honeys. Apis honey samples presented higher values of electric conductivity (284.00 μS cm^?1) than the obtained from the Jandaira honey samples (102.77 μS cm^?1) as well as a darker colour (26.67 mmPfund) when compared with Jandaira honey (7.00 mmPfund). The concentration of the glucose, fructose and sucrose was higher in the Apis honeys than in the Jandaira honey. The characteristics of the two types of honey were very different, highlighting the need of developing specific legislation for stingless bees' honey.     

Hepatoprotective Effects of the Honey of Apis cerana Fabricius on Bromobenzene‐Induced Liver Damage in Mice

Apis cerana honey (honey of Apis cerana Fabricius), widely distributed in the mountain areas of East Asia, has not been studied fully. The hepatoprotective activity of A. cerana honey was evaluated against bromobenzene‐induced liver damage in mice. In high dose, A. cerana honey can significantly alleviate liver injury, as is indicated by the depressed levels of serum alanine aminotransferase (ALT) (59.13%) and aspartate aminotransferase (AST) (79.71%), the inhibited malondialdehyde (MDA) content (63.30%), the elevated activities of superoxide dismutase (SOD) (73.12%) and glutathione‐Px (57.24%), and the decreased expression of Transforming growth factor β1 (51.83%) induced by bromobenzene (P < 0.05). The quantitative analysis of twelve major constituents (1 to 12) of A. cerana honey was executed by high performance liquid chromatography‐diode array detector. The results indicate that treatment with A. cerana honey can prevent bromobenzene‐induced hepatic damage in mice. Polyphenols might be the bioactive substances attributed to its antioxidant properties and intervention of oxidative stress.     

Authentication of beeswax (Apis mellifera) by high-temperature gas chromatography and chemometric analysis

Chemical characterization and authentication of beeswax of Apis mellifera was performed by high temperature capillary gas chromatography coupled to electron impact mass spectrometry or to flame ionisation detection and chemometric analysis. Many major components (>50) of beeswax, odd and even hydrocarbons, oleofin, palmitate, oleate and hydroxypalmitate monoesters were detected, and for the first time palmitate and oleate monoesters esterified with 1-octadecanol and 1-eicosanol are reported to be present in beeswax. Unsupervised pattern recognition procedures, cluster analysis and principal component analysis, were used to find data patterns and successfully differentiate authentic and paraffin adulterated beeswax based on the chemical profile obtained. Independent assessment of beeswax quality and performance of the unsupervised classification methods were performed using classical analytical parameters. The discrimination power of the chemometric unsupervised methods for detection of paraffin adulterated beeswax was superior to the discriminating power of classical analytical parameters. Using linear discriminant analysis, classification rules for authentic and paraffin adulterated beeswax samples were developed. The model was validated by leave-one-out cross validation and showed good recognition and prediction abilities, 100% and 99%, respectively.     

Biomedical Activity and Related Volatile Compounds of Thai Honeys from 3 Different Honeybee Species

This study investigated the effect of 3 factors (floral source, honeybee species, and postcollection processing) that influence the antibacterial activity, free radical reduction, and other biochemical compositions of different honey types typical of Thailand. Honey samples from 3 honeybee species (Apis mellifera, Apis cerana, and Apis dorsata) were obtained from 9 floral sources (longan, wild flower, lychee, coffee, sunflower, sesame, bitter bush, para‐rubber, and manuka as a control) in different regions of Thailand. These samples were evaluated for both their total and nonperoxide antibacterial activity against 10 human pathogens by agar incorporation technique. Honey samples were further analyzed to evaluate the capacity for free radical‐scavenging activity, total phenolic content, and the total flavonoid contents by the 2,2‐diphenyl‐1‐picrylhydrazyl assay, Folin–Ciocalteu method, and aluminum chloride colorimetric assay, respectively. Furthermore, the volatile organic compounds (VOCs) of Thai honey samples were investigated by headspace solid‐phase microextraction and gas chromatography‐mass spectrometry analysis. Findings of this study suggest a strong correlation between floral origin and honeybee species on one hand, and differences in %Brix, total acidity, protein content, antimicrobial activities, free radical reduction, phenolic, and flavonoid contents on the other hand. Moreover, VOCs of wild and coffee honey types were remarkably different, depending on the floral source. Both honeys contained characteristics of VOCs, some of which are involved in antibacterial and antioxidant activities.     

The botanical sources,entomological proteome and antibiotic properties of wild honey

We hypothesize that the wild honey (Apis dorsata honey (ADH), A. florea honey, A. laboriosa honey and A. andreniformis honey) had high antibacterial activity to defend against degradation. The entomological proteins, botanical sources and antibacterial activities of wild honey were determined via a label-free proteome, DNA metabarcoding and agar well diffusion assay. The results showed that there were significant differences among physico-chemical properties, proteins and plant sources of honey samples from different honey bee species. All the honey samples diluted to a 90% moisture content inhibited Staphylococcus aureus, Escherichia coli and Bacillus subtilis with the exception of 90% ADH. Honey solutions inhibited Chromobacterium violaceum, with exception of ADH and some other species honey with different moisture contents. The differences in antibacterial activity were attributed to differences in the botanical sources, entomological proteins of the honey. The species-specific proteins in honey from each species can be used for their identification     

Stingless bee honey: Quality parameters,bioactive compounds,health-promotion properties and modification detection strategies

BackgroundHoney is a natural product produced and marketed worldwide by stingless bees and Apis bees. Both these types of honey contain unique and distinct compounds of variable nutritional and biological importance. Stingless bee honey is popular for its distinct sweetness, mixed with an acidic taste, and fluid texture; it has higher added value than Apis mellifera honey. Due to the relatively low output of stingless bee honey compared to Apis mellifera honey, comprehensive data regarding the former is limited. This complex, natural product requires official, international methodologies and standards to be established to serve as a reference for quality control, to prevent adulteration, and to aid marketing purposes.Scope and approachThe article summarises the existing literature regarding the physicochemical parameters, chemical composition, bioactive constituents, biological properties, and modification detection strategies of honey originated from 478 honey samples from 66 different stingless bee species produced worldwide.Key findings and conclusionsStingless bee honey is one of the most complex natural foodstuffs. This type of honey quantitatively possesses a higher moisture content, greater acidity, a slightly lower level of total carbohydrates, and higher levels of antioxidant and biological activities than Apis mellifera honey. This review emphasises that stingless bee honey represents an important innovation for the food, pharmaceutical and cosmetic industries, due to its positive health effects and market potential.     

Storage-induced chemical changes in active components of honey de-regulate its antibacterial activity

To elucidate reasons for the observed variability in the antibacterial activity of honeys, we analysed a causal relationship between (a) honey floral sources and the activity and (b) the effect of honey storage on stability of compounds conferring this activity. Honeys from diverse floral sources were screened against Escherichia coli (ATCC 14948) and Bacillus subtilis (ATCC 6633) using the broth microdilution method. Among “active” honeys, 37% originated from buckwheat, 18% from clover and 12% from blueberry, indicating that these floral sources produced phytochemical(s) that inhibited bacterial growth. The stability of the putative phytochemical(s) was analysed in “active” honeys (MIC₉₀ 6.25% v/v) by measuring the activity every 3–6 months for a period of 1–3 years. A sharp decline in activity against both bacteria was observed in the first 3–6 months of storage. The decline coincided with major changes in chemical composition of honeys which included a significant change in colour (p < 0.0025), extremely significant change in concentration of UV-absorbing compounds (p < 0. 0001) and appearance of melanoidins. While these changes reduced E. coli sensitivity to honey, it rendered B. subtilis completely insensitive. Thus, the data indicates that the presence of phytochemical(s) conferring the antibacterial activity is sensitive to storage. The de-regulation of the antibacterial activity with the concomitant appearance of melanoidins suggests that the active phytochemical components might be sequestered into melanoidin aggregates, losing their function.     

Raw Millefiori honey is packed full of antioxidants

Total polyphenols, flavonoids and antioxidant power of raw honey samples from two of the most common Italian varieties, i.e., Millefiori and Acacia, were evaluated. Phenolic content, expressed as caffeic acid equivalents, ranged from 12.5 to 17.5 mg/100 g and from 3 to 11 mg/100 g in Millefiori and Acacia honeys, respectively. All Millefiori samples exhibited the highest flavonoid concentration being between 1.23 and 2.93 mg catechin equivalents (CE)/100 g honey. Total flavonoids in 100 g Acacia honeys were in the range of 0.45–1.01 mg CE. Acacia honeys had lower total antioxidant power, as assessed by ferric reducing/antioxidant power assay, than Millefiori. The relationship between phenolic content and antioxidant power was discussed. Comparative experimental analysis was performed with an artificial honey and processed honeys. Raw Millefiori honey is rich in both amount and variety of antioxidant substances, and its inclusion in the diet may be recommended to complement other polyphenol sources.     

Short communication: Viability of culture organisms in honey-enriched acidophilus-bifidus-thermophilus (ABT)-type fermented camel milk

The aim of this research was to monitor the survival during refrigerated storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in cultured dairy foods made from camel and, for comparison, cow milks supplemented with black locust (Robinia pseudoacacia L.) honey and fermented by an acidophilus-bifidus-thermophilus (ABT)-type culture. Two liters of dromedary camel milk and 2 L of cow milk were heated to 90°C and held for 10 min, then cooled to 40°C. One half of both types of milk was fortified with black locust honey at the rate of 5.0% (wt/vol), whereas the other half was devoid of honey and served as a control. The camel and cow milks with and without honey were subsequently inoculated with ABT-5 culture and were fermented at 37°C until a pH value of 4.6 was reached. Thereafter, the probiotic fermented milks were cooled to 15°C in ice water and were each separated into 18 fractions that were transferred in sterile, tightly capped centrifuge tubes. After 24 h of cooling at 8°C (d 0), the samples were stored at refrigeration temperature (4°C). Three tubes of all 4 products (i.e., fermented camel and cow milks with and without honey) were taken at each sampling time (i.e., following 0, 7, 14, 21, 28, and 35 d of storage), and the counts of characteristic microorganisms and those of certain spoilage microbes (yeasts, molds, coliforms, Escherichia coli) were enumerated. The entire experimental program was repeated twice. The results showed that addition of black locust honey at 5% to heat-treated camel and cow milks did not influence the growth and survival of starter streptococci during production and subsequent refrigerated storage of fermented ABT milks. In contrast, honey improved retention of viability of B. animalis ssp. lactis BB-12 in the camel milk-based product during storage at 4°C up to 5 wk. No spoilage organisms were detected in any of the samples tested in this study. In conclusion, supplementation of cultured dairy foods, especially those made from camel milk, with honey is recommended because honey is a healthy natural sweetener with a variety of beneficial microbiological, nutritional, and sensory properties.     

Changes in phosphorylation of myofibrillar proteins during postmortem development of porcine muscle

A gel-based phosphoproteomic study was performed to investigate the postmortem (PM) changes in protein phosphorylation of the myofibrillar proteins in three groups of pigs with different pH decline rates, from PM 1 to 24 h. The global phosphorylation level in the group with a fast pH decline rate was higher than that in the slow and intermediate groups at early PM time, but became the lowest at 24 h. The protein phosphorylation level of seven individual protein bands was only significantly (p < 0.05) affected by PM time, and two protein bands were subjected to a synergy effect between PM time and pH decline rate. A total of 35 non-redundant highly abundant proteins were identified from 19 protein bands; most of the identified proteins were sarcomeric function-related proteins. Myosin-binding protein C, troponin T, tropomyosin and myosin regulatory light chain 2 were identified in the highly phosphorylated protein bands with the highest scores. The results indicate that the phosphorylation pattern of myofibrillar proteins in PM muscle is mainly changed with PM time, but only to a minor extent influenced by the rate of pH decline, suggesting that the phosphorylation of myofibrillar proteins may be related to the meat rigor mortis and quality development.     

Production and characterization of mead from the honey of Melipona scutellaris stingless bees

Mead is a traditional alcoholic beverage obtained by fermenting must and can offer a solution to honey over‐production and a way of valorizing honey of lower quality. The purpose of this study was to produce and characterize mead with different levels of sugars and alcohol obtained from honey from Melipona scutellaris. The honey used for mead preparation was analysed in order to ensure that it met the required quality standards. It was found that the alcoholic content and volatile acidity were outwith the limits established by Brazilian law. Mead legislation is based on the product obtained from Apis mellifera (‘honey bee’) honey and these results indicate the need to re‐evaluate the standards established for this product in order to incorporate mead produced from honey from stingless bees of the genus Melipona. Copyright © 2018 The Institute of Brewing & Distilling     

Comparative proteomic analysis of Listeria monocytogenes exposed to enterocin AS-48 in planktonic and sessile states

Enterocin AS-48 is a cyclic peptide of great interest for application in food preservation and sanitation. In the present study, the proteome response of Listeria monocytogenes to purified enterocin AS-48 was studied under two different conditions: planktonic cells and sessile cells grown on polystyrene plates. Ten different proteins were differentially expressed in planktonic L. monocytogenes cells treated with 0.1 μg/ml enterocin AS-48 compared to the untreated controls. Overexpressed proteins were related to stress response (DnaK) or carbohydrate transport and metabolism, while underexpressed and unexpressed proteins were related to metabolism (such as glyceraldehyde-3-phosphate dehydrogenase, pyruvate oxidase, glutamate dehydrogenase or glutamate decarboxylase) or stress (GroEL). In the sessile state, L. monocytogenes cells tolerated up to 10 μg/ml bacteriocin, and the treated biofilm cells overexpressed a set of 11 proteins, some of which could be related to stress response (DnaK, GroEL), protein synthesis and carbohydrate metabolism, while glyceraldehyde-3-phosphate dehydrogenase was the only unexpressed protein. Some of the overexpressed proteins (such as elongation factor Tu and GroEL) could also be implicated in cell adhesion. These results suggest different cell responses of L. monocytogenes to enterocin AS-48 in the planktonic and in the sessile state, including stress response and cell metabolism proteins. While in the planktonic state the bacterium may tend to compensate for the cytoplasmic cell permeability changes induced by AS-48 by reinforcing carbohydrate transport and metabolism, sessile cells seem to respond by shifting carbohydrate metabolism and reinforcing protein synthesis. Stress response proteins also seem to be important in the response to AS-48, but the stress response seems to be different in planktonic and in sessile cells.     

Deterrent and insecticidal properties of bean seed (Phaseolus vulgaris L.) whole meal or protein extract incorporated into the diet of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae)

Callosobruchus maculatus, a pest that causes serious damage to chickpea Cicer arietinum, cannot develop in the seeds of Phaseolus or Vigna spp. which contain lectins. The insecticidal activity towards C. maculatus in these seeds is attributed both to lectins with specific affinity to N-acetylglucosamine, the major component of insect chitin, and to alpha-amylase inhibitors (lectin-like proteins). The insecticidal properties of bean meal or bean protein extracts from different sources towards different pest species are variable and need to be experimentally evaluated. The main objective of this study was to determine through a feeding trial on artificial chickpea seed the potential of bean seed meal from a wild bean Vigna caracalla, four varieties of Phaseolus vulgaris, and of a protein extract of P. vulgaris seed, to alter different life history traits of C. maculatus. The chickpea weevil was set up on artificial chickpea seeds containing different amounts of bean meal to observe the effects on female oviposition, percentage of development to adulthood and juvenile development time. These traits were combined in a composite index measuring the alteration of the multiplication rate of C. maculatus fed on artificial seed. The activity of lectin-like extracts was observed on chickpea artificial seed spiked with bean seed extract. Incorporation of bean flour at a rate of 10 and 20% in chickpea artificial seed significantly decreased C. maculatus female fecundity, percentage of adult emergence, and greatly increased the development time. Feeding trials with protein extracts of P. vulgaris reduced fecundity and survival of C. maculatus. Incorporation of 10% V. caracalla bean seed meal in chickpea artificial seed, reduced the multiplication potential of C. maculatus by over 90% showing that bean seed lectin extracts are worthy of further investigation for post-harvest infestation control.     

蜂蜜中原蜜黄酮类化合物HPLC图谱比较分析

利用高效液相色谱比较分析不同蜂种、不同产地、加工与否以及不同蜜源蜂蜜中原蜜黄酮类化合物的种类及含量差异。结果表明,意大利蜜蜂油菜原蜜和中华蜜蜂油菜原蜜总峰数和峰形都比较相似,意大利蜜蜂油菜原蜜的总黄酮含量比中华蜜蜂油菜原蜜总黄酮含量高。不同产地的意大利蜜蜂油菜原蜜的图谱峰形整体相似,总黄酮含量和黄酮类化合物种类大致相同。加工后的商品蜜总黄酮含量减少,黄酮类化合物的种类也减少,商品蜜的高效液相色谱图显示其黄酮类化合物的出峰时间靠前,主要密集分布在水溶性的分离相中。3个不同蜜源蜂蜜原蜜黄酮类化合物高效液相色谱图的总峰数和峰形都有较大差异,油菜原蜜、洋槐原蜜及柑橘原蜜的总峰数分别为65、58、70个,柑橘原蜜的总黄酮含量最高,达161.62μg/100 g。单花蜂蜜原蜜黄酮类化合物的高效液相色谱图峰形具有一定的特异性,可以作为指纹图谱用于蜂蜜蜜源的鉴定。     

Production and partial purification of proteases from Aspergillus oryzae grown in a medium based on whey protein as an exclusive nitrogen source

Several attempts have been made to incorporate whey proteins into curd to increase cheese yield. For some types of cheese, degradation of whey proteins that have been incorporated into the curd would be required to obtain acceptable flavor and texture. On the basis of the high potential for protease synthesis in Aspergillus oryzae, sodium nitrate as a nitrogen source in a minimal medium for fungi, known as Czapek-Dox medium, was replaced with whey protein isolate to induce the protease to hydrolyze whey protein using A. oryzae AHU7146. A solid-phase medium adjusted to pH 6 was suitable for this purpose when incubation was carried out at 25°C for 2 wk. The application of column chromatography enabled the resolution of 3 proteolytic components (1, 2, and 3). With respect to optimal temperature and zymographic analysis, component 1 was similar to component 3. In contrast, component 2 was less abundant than the other components and exhibited activity in the alkaline pH region. The degradation of β-lactoglobulin and α-lactalbumin in whey protein isolate solution by the crude enzyme was primarily attributed to the action of components 1 and 3, based on HPLC analysis and the N-terminal amino acid sequences; however, zymography demonstrated evident proteolysis due to component 2. Because heat-denatured whey protein aggregates were digestible by the crude enzyme, the proteolytic system from A. oryzae has the potential as an additive to stimulate the ripening of cheese enriched with whey protein.     

A novel glycoprotein from mushroom Hypsizygus marmoreus (Peck) Bigelow with growth inhibitory effect against human leukaemic U937 cells

This study was to isolate the anti-leukaemic component from edible mushroom Hypsizygus marmoreus (Peck) Bigelow. Crude protein was extracted from the basidioma, and then purified with DEAE-Sepharose CL-6B ion exchange chromatography followed by Sephacryl S-300 gel filtration. A protein which exerted high growth inhibitory effect on human leukaemic U937 cells and sufficient toxicological safety on normal human white blood cells was isolated and named HM-3A. Electrophoresis showed HM-3A approximately 52 kDa in size. N-terminal analysis found the amino acid sequence ATTQWKTSAA and confirmed HM-3A a novel protein. High-performance anion-exchange column chromatography revealed HM-3A a glycoprotein with galactose as the major monosaccharide. Haemagglutination assay proved it non-lectin. We suggest that HM-3A is worth further investigation for antitumour use.     

Characterization of Callosobruchus chinensis (L.) resistance in Vigna umbellata (Thunb.) Ohwi & Ohashi

Resistance to azuki bean weevil, Callosobruchus chinensis, was studied in a series of field and laboratory experiments in two accessions of rice bean (Vigna umbellata (Thunb.) Ohwi & Ohashi), one accession of black gram (V. mungo (L.) Hepper), and one accession of mungbean, (V. radiata (L.) Wilczek). Weevil damage to immature pods of the rice bean accessions, ‘Menaga’ and ‘Miyazaki’, was significantly less than to the susceptible mungbean, VC1973A. In mature pods, the pest damage to the pod wall of Menaga was significantly higher than to VC1973A, whereas the damage to Miyazaki was similar to VC1973A. Seeds within the pods of both rice bean accessions were resistant no matter when the pods were harvested. When the insects were exposed directly on dry seeds, both rice bean accessions and a black gram accession VM2164 were resistant to them. In artificial seeds made by mixing flour of the individual resistant Vigna accessions with VC1973A and subsequently exposed to bruchid oviposition, the higher the quantity of resistant Vigna flour the lower the number of bruchids that emerged from such seeds. No bruchids emerged from artificial seeds containing crude starch fraction from the three resistant Vigna accessions when such seeds were exposed to bruchid infestation, whereas many insects emerged from the seeds containing starch of VC1973A or flour of VC1973A alone. In artificial seeds made by mixing crude protein fractions of the three resistant Vigna accessions with flour of VC1973A, as the concentration of protein increased the number of C. chinensis adults that emerged decreased. Fractionation of crude proteins into acetone-precipitable proteins and peptide and amino acid portions resulted in the loss of antibiosis effect. Artificial seeds made from purified starch-polysaccharides fraction, however, exhibited antibiosis effects if prepared from the rice bean seed of Menaga and Miyazaki but not if made from the black gram seed, VM2164.     

Isolation and characterization of a novel analyte from Bacillus subtilis SC-8 antagonistic to Bacillus cereus

In this study, an effective substance was isolated from Bacillus subtilis SC-8, which was obtained from traditionally fermented soybean paste, cheonggukjang. The substance was purified by HPLC, and its properties were analyzed. It had an adequate antagonistic effect on Bacilluscereus, and its spectrum of activity was narrow. When tested on several gram-negative and gram-positive foodborne pathogenic bacteria such as Salmonella enterica, Salmonella enteritidis, Staphylococcus aureus, and Listeria monocytogenes, no antagonistic effect was observed. Applying the derivative from B. subtilis SC-8 within the same genus did not inhibit the growth of major soybean-fermenting bacteria such as Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens. The range of pH stability of the purified antagonistic substance was wide (from 4.0 to >10.0), and the substance was thermally stable up to 60 °C. In the various enzyme treatments, the antagonistic activity of the purified substance was reduced with proteinase K, protease, and lipase; its activity was partially destroyed with esterase. Spores of B. cereus did not grow at all in the presence of 5 μg/mL of the purified antagonistic substance. The isolated antagonistic substance was thought to be an antibiotic-like lipopeptidal compound and was tentatively named BSAP-254 because it absorbed to UV radiation at 254 nm.     

Protein isolates from two Mediterranean legumes: Lathyrus clymenum and Lathyrus annuus. Chemical composition,functional properties and protein characterisation

Protein isolates were analysed from two Mediterranean legumes, Lathyrus clymenum and L. annuus. Protein isolates were prepared by alkaline extraction, including sodium sulphite and acid precipitation of Lathyrus proteins at their isoelectric point (pH 4.5). The percentage of proteins recovered from L. annuus and L. clymenum flours during the preparation of the protein isolates was around 60%. Chemical composition, nutritional parameters, main functional properties and protein composition of Lathyrus protein isolates were studied. L. annuus and L. clymenum protein isolates contained 81.07% and 82.4% of proteins, respectively, and they have a balanced content of essential amino acids, except for sulphur amino acids, with respect to the FAO pattern. The in vitro protein digestibility increased in the protein isolates to 93% and 95% in L. annuus and L. clymenum, respectively. Functional properties were similar to those observed in other legumes protein isolates. These results confirm the interest of local crops as sources of high value protein products obtained after convenient protein extraction procedures and the removal of antinutritional components. These high added value protein isolates are of interest for the food industry and for the revalorisation of L. annuus and L. clymenum favouring the bioconservation of Lathyrus.