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[Influence of hydrothermic and enzymatic treatments on the nutritive value of Faba beans: in vitro study].

In order to improve the digestibility of the Faba bean flour an hydrothermic enzymic and fermentation treatment has been thought out (US patent 395 8015, Ets Ury, M. Gay). This study aims at stemming out the nutritionnal and structural repercusions of this treatment on the Faba proteins. The nitrogen distribution is deeply changed : total nitrogen increase (47%), water soluble nitrogen decrease (41%), water soluble non-protein nitrogen increase. The treatment can enrich the flour in nitrogen but the proteins supplement is insoluble in water. The treatment changed the electrophoretic behaviour of flour proteins letting disappear precipitable pH 4,5 proteins. Amino acid composition is slightly changed, however there is an increase of lysine and methionine (15 et 25%) and a decrease of cysteine (28%). The chemical score shows sulfur amino acids and tryptophan deficiency. Enzymatic (pepsine and pancreatine mixture) liberation of all amino acids is low and is not improved by the treatment but for the lysine. Cell proteins supplement, which appeared during the treatment (61%), water insoluble, could improve nutritive value of the flour if a more complete destruction of the cell walls permitted their liberation and their solubilization.

Amino Acids↗

A single rep protein initiates replication of multiple genome components of faba bean necrotic yellows virus, a single-stranded DNA virus of plants.

Faba bean necrotic yellows virus (FBNYV) belongs to the nanoviruses, plant viruses whose genome consists of multiple circular single-stranded DNA components. Eleven distinct DNAs, 5 of which encode different replication initiator (Rep) proteins, have been identified in two FBNYV isolates. Origin-specific DNA cleavage and nucleotidyl transfer activities were shown for Rep1 and Rep2 proteins in vitro, and their essential tyrosine residues that catalyze these reactions were identified by site-directed mutagenesis. In addition, we showed that Rep1 and Rep2 proteins hydrolyze ATP, and by changing the key lysine residue in the proteins' nucleoside triphosphate binding sites, demonstrated that this ATPase activity is essential for multiplication of virus DNA in vivo. Each of the five FBNYV Rep proteins initiated replication of the DNA molecule by which it was encoded, but only Rep2 was able to initiate replication of all the six other genome components. Furthermore, of the five rep components, only the Rep2-encoding DNA was always detected in 55 FBNYV samples from eight countries. These data provide experimental evidence for a master replication protein encoded by a multicomponent single-stranded DNA virus.

Adenosine Triphosphatases↗

Dose-independent effect of misonidazole in fractionated irradiations of hypoxic Vicia faba bean roots.

The radiosensitization of 5 mM misonidazole (Miso) was measured in Vicia faba bean roots with regimens of single, three, six and twelve fractions of 250 kVp X-rays. To inhibit cell division, the beans were kept at a constant temperature of 3.5 degrees C during irradiation and between fractions that were spaced 24 hours apart. The doses in various regimens were graded such that they ranged between 27 and 350, and 42 and 513 cGy per fraction in Miso-treated and non-treated regimens, respectively, under hypoxia. The sensitivity enhancement ratio (s.e.r.) was constant throughout the dose range employed with an average value of 1.62. The s.e.r. increased to 2.3 when measured with single doses at 19 degrees C. It is concluded that the s.e.r. is dose-independent and that temperature enhances the effectiveness of the drug.

Dose-Response Relationship, Drug↗

Analysis of six DNA components of the faba bean necrotic yellows virus genome and their structural affinity to related plant virus genomes.

Faba bean necrotic yellows virus (FBNYV) has a multicomponent circular ssDNA genome. In addition to a previously described genome component (C1) coding for a replicase-associated protein (Rep), five further components (C2 to C6) have now been identified. Each of the six components is about 1 kb in size, contains one major open reading frame (ORF) in the virion sense with a TATA box and polyadenylation signal, and has a noncoding region containing a highly conserved sequence possibly forming a stem-loop structure. Similar to C1, C2 encodes another putative Rep of 33.1 kDa, which is closely related to the Rep of banana bunchy top virus (BBTV). Based on bacterial expression and immunoblot analysis, the ORF of C5 encodes the capsid protein (CP) with a deduced molecular mass of 19 kDa. The FBNYV CP shares the highest amino acid (aa) identity (56.2%) with that of subterranean clover stunt virus (SCSV). The ORF of C4 potentially codes for a hydrophobic protein which appears to be structurally and functionally similar to the BBTV-C4 and SCSV-C1 proteins. No protein sequence similarities were found in databases for the C3 and C6 ORFs of FBNYV. FBNYV is clearly distinct from any known virus but is taxonomically related to BBTV and SCSV.

Amino Acid Sequence↗

Faba bean necrotic yellows virus (genus Nanovirus) requires a helper factor for its aphid transmission.

Purified faba bean necrotic yellows virus (FBNYV; genus Nanovirus) alone is not transmissible by its aphid vector, Acyrthosiphon pisum, regardless of whether it is acquired from artificial diets or directly microinjected into the aphid's hemocoel. The purified virus contains all of the genetic information required for its infection cycle as it readily replicated in cowpea protoplasts and systemically infected Vicia faba seedlings that were biolistically inoculated using gold particles coated with intact virions or viral DNA. The bombarded plants not only developed the typical disease syndrome, thus indicating that FBNYV is the sole causal agent of the disease, but also served as a source from which the virus was readily acquired and transmitted by A. pisum. The defect of the purified virus in aphid transmissibility suggests that FBNYV requires a helper factor (HF) for its vector transmission that is either nonfunctional or absent in purified virus suspensions. The requirement for an HF was confirmed in complementation experiments using two distinct isolates of the virus. These experiments revealed that aphids transmitted the purified virus isolate from artificial diets only when they had fed previously on plants infected with the other FBNYV isolate. Also, microinjected FBNYV, which persisted to the same extent in A. pisum as naturally acquired virus, was transmissible when aphids had acquired the HF from infected plants. This suggests that one of the functions of the HF in the transmission process is to facilitate virus transport across the hemocoel-salivary gland interface.

Amino Acid Sequence↗

Physiochemical characteristics of flours of faba bean as influenced by processing methods.

Wet- and dry-processing with and without heating treatments were used to dehull faba beans for preparation of flours from the cotyledons. Flour qualities were assessed by levels of tannin and trypsin inhibitor and other measures of proximate composition. High roasting temperature and shorter process time that improved the recovered cotyledon yields were verified significant by path analysis methodologies. In turn, the higher cotyledon recoveries correlated with higher protein levels and inversely with the measured tannin levels. Ash reductions were correlated to the wet processing options in hull removal while reductions in the insoluble dietary fiber were notably influenced by level of heating temperatures applied. Trypsin inhibitor levels ranged from 42 to 56% of the original with reductions tied to applications of wet and/or heat processing in each case improved by increased durations and temperatures of treatments.

Chemical Phenomena↗

Changes in interfacial behaviour, emulsifying and foaming properties of faba bean legumin after modification with dimethylsuberimidate.

The effect of a rising rigidity and surface hydrophobicity of the 11S storage protein from faba beans--legumin--induced by chemical modification with dimethylsuberimidate (DMS) on some surface functional properties was studied. Short-time adsorption kinetics using a droplet-volume tensiometer, pressure transformation and desorption behaviour of monolayer using a film balance, and emulsifying and foaming properties were determined to characterize surface activity and interfacial film forming behaviour. Tensio-active properties at the air-water interface, i.e. decay in surface tension and pressure transformation in monolayer, were improved by modification. However, a decrease in emulsifying activity, foam capacity and foam expansion after modification of the legumin points to an overall deterioration of energy-induced film forming behaviour. The results support the view that surface activity is generally governed more by molecular flexibility than by surface hydrophobicity.

Dimethyl Suberimidate↗

Structural and functional changes of faba bean legumin during super-limited tryptic hydrolysis.

The influence of a super-limited tryptic hydrolysis on physicochemical and surface functional properties of faba bean legumin has been studied using size-exclusion HPLC, SDS-PAGE, UV and fluorescence spectroscopy, fluorescence probe techniques, surface tension measurements as well as determination of emulsifying activity index (EAI) and emulsion droplets diameter (D). The extent of legumin hydrolysis comprised the range between about 14 and 60 split peptide bonds per molecule resulting in a stepwise decrease of legumin molecular weight to 240 kDa (legumin-T) via discrete intermediates with characteristic subunit patterns. These changes are accompanied by an increase in the surface hydrophobicity and the exposure of aromatic chromophores. No differences were found in the surface tension between the variously hydrolyzed legumin samples. Best emulsifying properties (highest EAI and lowest D values) were attained after a rather low tryptic hydrolysis (about 30 split peptide bonds per mol). Further hydrolysis impaired the emulsifying parameter which were, however, higher (EAI) or lower (D) than those for native legumin.

Amines↗

Ten distinct circular ssDNA components, four of which encode putative replication-associated proteins, are associated with the faba bean necrotic yellows virus genome.

Four further circular ssDNA components (C7-C10), about 1 kb in size and structurally similar to the previously described components (C1-C6) found associated with a Syrian (Sy) isolate of faba bean necrotic yellows virus (FBNYV), have been identified. Similar to C1 and C2, two of the new components (C7 and C9) encode putative replication-associated (Rep) proteins of 33.2 and 32.7 kDa, respectively, the former of which is 90% identical to the C10 Rep protein of milk vetch dwarf virus (MDV). C8 encodes a putative protein (17.4 kDa) whose function is unknown, but which is highly conserved between FBNYV and the other nanoviruses MDV, subterranean clover stunt virus and banana bunchy top virus. The putative protein (19.7 kDa) encoded by C10 contains an LXCXE motif, which is also present in the homologues of its relatives, suggesting that they may all interact with plant retinoblastoma-like proteins. Sequence information for seven components of an Egyptian (Eg) FBNYV isolate indicated that six of them share > 96% identity with FBNYV-Sy. However, the C1 Rep protein of FBNYV-Eg was only 63.5% identical to that of FBNYV-Sy, but was 88.3 % identical to the MDV-C2 Rep protein. We conclude that the FBNYV genome consists of seven to ten components, six of which encode non-Rep proteins. All ten components of the FBNYV genome, except the Rep components C2 and C9, had closely related counterparts in the MDV genome. In spite of this similarity, FBNYV and MDV appear to be distinct virus species.

Amino Acid Sequence↗

[In vitro digestibility of original and modified protein isolate from faba beans].

Protein isolate from Vicia faba (API) and 18 different partial hydrolysates of API as well as plasteins produced from them were submitted to an in vitro digestion with pepsin and pancreatic proteases at their specific pH-optimum. By means of this technique a final value of hydrolysis (49,8 +/- 3,0)% was obtained with all investigated samples, in spite of the different intensity of the preliminary hydrolysis. Two samples with a degree of preliminary hydrolysis of 35% showed with 55% and 56%, respectively, a significantly higher final value. From the results may be assumed that by means of partial hydrolysis and plastein formation only such peptides will be formed which are again hydrolyzable in vitro.

Fabaceae↗

Radiobiological intercomparison of clinical neutron beams for growth inhibition in Vicia faba bean roots.

Relative biological effectiveness (RBE) and oxygen enhancement ratio (OER) values of different neutron beams produced at the variable energy cyclotron "Cyclone" of Louvain-la-Neuve (Belgium) were determined. The neutrons were obtained by bombarding a beryllium target with 34-, 45-, 65-, or 75-MeV protons or with 50-MeV deuterons. The biological system was growth inhibition in Vicia faba bean roots. Taking the p(65) + Be neutron beam as a reference, RBE values were found equal to 1.36 +/- 0.2, 1.20 +/- 0.1, 1.00 (ref), 0.98 +/- 0.1, and 1.18 +/- 0.1, respectively; the doses corresponding to 50% growth inhibition were 0.39, 0.44, 0.53, 0.54, and 0.45 Gy. For the same beams, OER values were found equal to 1.55 +/- 0.1, 1.38 +/- 0.1, 1.29 +/- 0.1, 1.41 +/- 0.1, and 1.60 +/- 0.2, respectively.

Growth↗

Isolates from faba bean and soybean with lowered content of phytic acid and activity of the trypsin inhibitors.

For obtaining protein isolates, water, whey, and waste effluents from a potato processing plant were used as extraction solvents. Isolates obtained on laboratory conditions were subjected to chemical and organoleptic tests. It was shown that proper steering of the processing allowed for obtaining isolates with lower phytic acid content and decreased activity of the trypsin inhibitors, although the efficiency of the process decreased also. Modified processing was more efficient for elimination of the phytic acid from soybean isolates compared to faba bean. It was also shown that isolate neutralization considerably improved their solubility and decreased the activity of trypsin inhibitors.

Chemical Phenomena↗

Bioavailability to rats and toxicity in mice of carbofuran residues bound to faba beans.

14C-carbofuran penetrated readily into seeds of Vicia faba and the rate of penetration was found to be dose dependent. The percentage of bound residues was generally low and did not exceed 3% of the applied dose. When the bound residues were fed to rats 46% of the radioactivity was eliminated via CO2 and urine, while tissues contained 25%. Carbofuran phenol and 3-hydroxy carbofuran represented the main metabolites in the urine. These data indicate that bean-bound carbofuran residues are highly bioavailable to rats. Feeding mice with bound carbofuran residues for 90 days led to inhibition of erythrocyte cholinesterase activity after 30 days (35-40%) while the plasma enzyme remained unaffected. Serum transaminases and blood urea nitrogen were significantly elevated, indicating injury to hepatic and renal structures. The results strongly suggest that the bound residues can induce adverse biological effects in mice.

Animals↗

[Structure of the photosynthetic apparatus of the bean Vicia faba L. when treated with 6-benzylaminopurine].

The level of endogenous cytokinins changed with growth and development of faba bean (Vicia faba L.) leaves. Typical of juvenile leaves, amounting to 25% of the final leaf size (Smax) was a low content of these plant hormones. The level of cytokinins increased in growing leaves (50% of Smax) and decreased in the leaves that stopped growing. The content of cytokinins in senescent leaves dropped considerably. Exogenous treatment with 6-benzylaminopurine (BAP) had no effect on the structure of terminal phloem; however, it stimulated elongation of mesophyll cells; increases in the area and thickness of leaf blade, amount of photosynthetic pigments, and assimilation potential; and delayed senescence of leaves and defoliation, thereby increasing biomass of the aboveground plant part. It was inferred that BAP had a potential for induction of photosynthetic apparatus development and increase in the yield of faba bean green mass.

Adenine↗

Hormone-related, muscle-specific changes in protein metabolism and fiber type profile after faba bean intake.

Male growing Wistar rats were fed, over 15 days, isoenergetic (16.72 +/- 0.49 MJ) and isoproteic (11%) diets containing either lactalbumin or raw Vicia faba L. (Vf) as the sole source of protein. Compared with pair-fed controls (PF), soleus muscles of Vf-fed rats showed increased (P < 0.05) synthesis and breakdown rates. In addition, the soleus of Vf-fed rats displayed a decrease (P < 0.05) in type I and an increase (P < 0.01) in type IIc fibers compared with that of PF animals. On the contrary, extensor digitorum longus muscles of both Vf-fed and PF rats showed an increase (P < 0.01) in type I and a reduction (P < 0.05) in type IIb fibers together with a decrease (P < 0.05) in the cross-sectional area of the latter fibers. Vf-fed rats exhibited a significant decrease in serum insulin (P < 0.05) and thyrotropin (P < 0.01) levels, together with an increase in plasma glucagon (P < 0.05) and 3,5,3'-triiodothyronine (P < 0.01) concentrations, compared with the PF group. Both Vf-fed and PF rats experienced an increase in corticosterone concentrations (P < 0.01 vs. control; P < 0.05 vs. PF). The muscle-specific changes in both protein metabolism and fiber type composition may partly depend on the hormonal changes that were observed after Vf intake.

Amino Acids↗