Design and Characterization of a Novel Fermented Beverage from Lentil Grains

The experimental activities carried out in this study aimed at designing a lentil-based beverage rich in soluble and digestible proteins. In order to extract soluble proteins, lentil grains were soaked in water overnight, blended, treated with proteolytic enzymes and fermented with Lactobacillus strains. Protein enzymatic hydrolysis, carried out with four commercial food grade enzyme preparations, showed that bromelin, at the enzyme to substrate ratio of 10%, was the best solution to produce this novel beverage. Even though the seven Lactobacillus strains were all able to ferment aqueous extract within 24 h, L. acidophilus ATCC 4356, L. fermentum DSM 20052 and L. paracasei subsp. paracasei DSM 20312 showed the highest growth rate and the lowest pH values. In fermented lentil-based beverages, the antinutritional factor phytic acid decreased up to 30%, similarly, the highest reduction in raffinose oligosaccharides content reached about 12% the initial concentration. It is worthy of note that the viable density of all strains remained higher than 7 log cfu/mL after 28 days of cold storage. The results here reported show for the first time the possibility to obtain a probiotic lentil-based beverage rich in soluble proteins, peptides and amino acids with low content in main antinutritional factors.

Expression of Oryza sativa galactinol synthase gene in maize (Zea may L.)

Galactinol synthase (GolS) is a key biological catalyst for the synthesis of the raffinose oligosaccharides (RFOs) which play important roles in abiotic stress adaptation of plants, especially drought tolerance. GolS gene has been isolated on a variety of plants in order to create material resources for generating transgenic plants resistant to adverse environmental factors. In our previous research, we have isolated a GolS gene from drought stress cDNA library of Oryza sativa L. Moctuyen (named OsGolS). In this study, the expression vector pCAM-Rd/OsGolS carrying the isolated OsGolS gene under the control of stress-inducible Rd29A promoter was constructed and introduced into Agrobacterium tumefaciens LBA4404, which was used for maize transformation. PCR and Real-time PCR assay indicated that transgene was integrated in the genome of the regenerated Zea mays plants. Reverse transcription-PCR showed that the OsGolS was transcribed into mRNA in Zea mays and was highly expressed. These results provide a basis for the study of the function of OsGolS in drought responses and for the development of drought stress tolerant crops.

Effect of Lactic Acid Bacteria Starter Cultures on Vitamin and Oligosaccharide Composition of Milk Extracted from Three Common Bean (Phaselous Vulgaris L) Varieties

Fermented foods have in recent times attracted consumer interest mainly due to perceived health benefits of probiotic microorganisms. This study characterized changes in the concentrations of selected B-complex vitamins and oligosaccharides of common bean milk during fermentation by a common dairy starter culture, YF L-903 (Streptococcus thermophilus + Lactobacillus Bulgaricus subs Debulgaricus), and three probiotic cultures namely ABT (Lactobacillus acidophilus La-5 + Bifidobacterium animalis Bb-12 + Streptococcus thermophilus), Yoba (Lactobacillus rhamnosus yoba + Streptococcus thermophilus), and Yoba Fiti (Lactobacillus rhamnosus GR1 + Streptococcus thermophilus). Bean milk was prepared from three common bean varieties. It was found that, apart from thiamine (vitamin B1) and riboflavin (vitamin B2), fermentation with each of the mixed cultures caused significant increase in the vitamin B complex. Significant reductions (p<0.05) in the oligosaccharides concentration of the bean milks were observed upon fermentation. Highest reduction in the oligosaccharide sugars of 77.8% was found in milk from pinto bean variety fermented with ABT culture. These findings suggest that LAB probiotic cultures have a potential for improving biosynthesis of vitamins and removal of the verbascose, stachyose and raffinose oligosaccharides, thus making the product more digestible and the nutrients more bioavailable.

Biosynthesis of raffinose family oligosaccharides and galactosyl pinitols in developing and maturing seeds of winter vetch (Vicia vlllosa Roth.)

Changes in the accumulation of two types of α-D-galactosides: raffinose family oligosaccharides and galactosyl pinitols were compared with changes in the activities of galactosyltransferases during winter vetch (<em>Vicia villosa</em> Roth.) seed development and maturation. Occurrence of galactinol and raffinose in young seeds and changes in activities of galactinol synthase and raffinose synthase during seed development indicated that formation of raffinose oligosaccharides (RFOs) preceded synthesis of galactopinitols. Although transfer of galactose residues into raffinose oligosaccharides increased as seeds were maturing, at late stages of seed maturation the accumulation of galactopinitols was preferred to that of RFOs. In the present study, activities of enzymes transferring galactose moieties from galactinol to D-pinitol forming galactopinitol A, and further transfer of galactose moieties from galactinol to mono- and di-galactopinitol A were detected throughout seed development and maturation. This is a new observation, indicating biological potential of winter vetch seeds to synthesize mono-, di- and tri-galactosides of D-pinitol in a pathway similar to RFOs. The pattern of changes in activities of stachyose synthase and enzymes synthesizing galactopinitols (named galactopinitol A synthase and ciceritol synthase) suggests that formation of stachyose, mono- and di-galactopinitol A (ciceritol) is catalyzed by one enzyme. High correlation between activities of verbascose synthase and enzyme catalyzing synthesis of tri-galactopinitol A from galactinol and ciceritol (named tri-galactopinitol A synthase) also suggests that biosynthesis of both types of tri-galactosides was catalyzed by one enzyme, but distinct from stachyose synthase. Changes in concentrations of galactosyl acceptors (sucrose and D-pinitol) can be a factor which regulates splitting of galactose moieties between both types of galactosides in winter vetch seeds.

Purification and characterization of an alpha-galactosidase from Aspergillus fumigatus

Aspergillus fumigatus secreted invertase (beta-fructofuranosidase) and alpha-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). alpha-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtration and two ion exchange chromatographies and showed Mw of 54.7 kDa. The purified enzyme showed maximal activity against p-nitrophenyl-alpha-D-galactopyranoside (pNPGal) at pH 4.5-5.5 and 55 °C, and retained about 80% of the original activity after incubation for 90 minutes at 50ºC. The KM for pNPGal was 0.3 mM. Melibiose was hydrolyzed by the enzyme but raffinose was very poor substrate.