Anna Jakubczyk

Effect of abiotic elicitation on main health-promoting compounds, antioxidant activity and commercial quality of butter lettuce (Lactuca sativa L.)

The study presents changes in the phytochemical levels, antiradical activity and quality of lettuce caused by different chemical elicitors: arachidonic acid (AA), jasmonic acid (JA), and abscisic acid (ABA). The application of 1 μM and 100 μM JA induced an increase in the concentration of phenolic compounds, including flavonoids and phenolic acids. Flavonoid levels were also increased after treatment with 100 μM AA and ABA. Some of the elicitor concentrations used also caused an increase in the levels of other phytochemicals, such as chlorophyll a (1 μM and 100 μM AA, 50 μM ABA); chlorophyll b (100 μM AA); carotenoids (100 μM AA, 1 μM JA and 100 μM ABA) and vitamin C (100 μM AA, 100 μM JA). The highest antiradical activity was noted after treatment with 100 μM AA, 100 μM JA. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) scavenging ability was positively and significantly correlated with flavonoid, chlorophyll and carotenoid levels. These results may suggest that the antiradical activity of lettuce was determined not only by phenolics, but also by other bioactive compounds. Elicitation did not change the sensory quality of lettuce. Therefore, treatment with elicitors could be a useful tool for improving the health-promoting qualities of lettuce without the loss of sensory quality.

Biologically active peptides obtained by enzymatic hydrolysis of Adzuki bean seeds

This study investigated the antioxidant and antihypertensive activities of peptides obtained from protein fractions of Adzuki bean seeds. Peptides were obtained by the use of hydrolytic enzymes in vitro under gastrointestinal conditions. A determination was made of the activity of the peptide inhibitors of the angiotensin I converting enzyme (ACE), and the antiradical and ion chelating activity of peptides from different protein fractions. The highest peptide levels after the absorption process (<7 kDa) were noted in the albumin fraction (50.69 μg/ml). Furthermore, it was found that peptides from the prolamin fraction were characterised by the highest antiradical activity and ACE inhibitory activity (IC50=0.17 mg/ml). Peptides obtained from the globulin fraction showed the highest ability to chelate iron ions, and peptides from the glutelin fraction were characterised as being the most effective in the chelation of copper ions.

The impact of fermentation and in vitro digestion on formation angiotensin converting enzyme (ACE) inhibitory peptides from pea proteins.

Pea seeds were fermented by Lactobacillus plantarum 299v in monoculture under different time and temperature conditions and the fermented products were digested in vitro under gastrointestinal conditions. After fermentation and digestion ACE inhibitory activity was determined. In all samples after fermentation no ACE inhibitory activity was noted. Potentially antihypertensive peptides were released during in vitro digestion. The highest DH (68.62%) were noted for control sample, although the lowest IC50 value (0.19 mg/ml) was determined for product after 7 days fermentation at 22 °C. The hydrolysate characterised by the highest ACE inhibitory activity was separated on Sephadex G10 and two peptides fractions were obtained. The highest ACE inhibitory activity (IC50=64.04 μg/ml) for the first fraction was noted. This fraction was separated by HPLC and identified by LC-MS/MS and the sequence of peptide derived from pea proteins was determined as KEDDEEEEQGEEE.

Stevia Rebaudiana Bert. Leaf Extracts as a Multifunctional Source of Natural Antioxidants

The aim of the presented study was to characterize the content and biological activity of extracts prepared from dried Stevia rebaudiana leaves with potential application in the food or cosmetic industry. Aqueous (A), ethanolic (E) and glycol-aqueous (GA) extracts were analyzed for the content of polyphenols and proteins, showing that the highest amount of phenols (15.50 mg/g) and flavonoids (3.85 mg/g) contained GA. All extracts contained significant amount of protein (69.40–374.67 mg/g). Between analyzed stevia extracts (HPLC) GA contained the highest amount of polyphenols, especially ferulic (5.50 mg/g) and rozmaric (4.95 mg/g) acids derivates. The highest antiradical activity against DPPH• and ABTS•+ was noted for GA and E (IC50 = 0.38 and 0.71 µg flavonoids/mL). The highest ability to chelate Fe2+ was observed for E (IC50 = 2.08 µg flavonoids/mL). Stevia extracts were also analyzed for their cytotoxicity and fibroblast irritation potential in vitro. E and GA were the most cytotoxic and irritating, probably due to the high content of biologically active phytochemicals. On the other hand, a extract was the most tolerable by the cells. To summarize, the presented study evaluated the potential application of A, E and GA stevia extracts as natural source of antioxidants in the food and cosmetic industry.

Angiotensin I converting enzyme inhibitory peptides obtained after in vitro hydrolysis of pea (Pisum sativum var. Bajka) globulins.

Pea seeds represent a valuable source of active compounds that may positively influence health. In this study, the pea globulins were digested in vitro under gastrointestinal condition and potentially bioaccessible angiotensin I converting enzyme (ACE) inhibitory peptides were identified. The degree of hydrolysis after pepsin, 14.42%, and pancreatin, 30.65%, were noted. The peptides with the highest ACE inhibitory properties were separated using ion exchange chromatography on DEAE-cellulose. Thirteen peptides fractions were obtained but only four showed potential antihypertensive properties. The highest inhibitory activity was determined for the fraction F8 (IC50 = 0.0014 mg/mL). This fraction was separated on Sephadex G10 and two peptide fractions were obtained. The peptides fraction (B) with the highest ACE inhibitory activity (IC50 = 0.073 mg/mL) was identified by ESI-MS/MS. The sequences of ACE inhibitory peptides were GGSGNY, DLKLP, GSSDNR, MRDLK, and HNTPSR. Based on Lineweaver-Burk plots for the fraction B, the kinetic parameters as K m, Vmax, and K i and mode of inhibition were determined. This fraction belongs to uncompetitive inhibitor of ACE activity. The seeds of pea are the source of precursor protein, which releases the ACE inhibitory peptides as a result of enzymatic hydrolysis.

Enhancement of yield, nutritional and nutraceutical properties of two common bean cultivars following the application of seaweed extract (Ecklonia maxima)

In the present study, application of Ecklonia maxima extract (Kelpak SL – a water soluble concentrate) was optimized and its impact on yield, nutraceutical and nutritional potential of Phaseolus vulgaris L. (var. Aura and Toska) was measured. The study was carried out in 2012 and 2013 in Poland. During the growing season, 0.2% and 0.4% solution of Kelpak SL was applied by single and double spraying of plants. These four treatments with Kelpak SL were compared with the control, where no biostimulator was applied. Kelpak SL treatments stimulated the yield of both cultivars studied. The application of E. maxima extract had no effect on the content of starch, free sugars or proteins in seeds of either of the tested cultivars. The highest level of phenolics was found for double sprayed Toska plants. All the tested variants of Kelpak SL application significantly increased the content of anthocyanins in the seeds. Also, both the reducing power and antiradical ability of Aura seeds were elevated in all the studied treatments. E. maxima extract is a natural, environmentally friendly and safe preparation increasing the yield and nutraceutical quality of beans without any negative effect on their nutritional quality.

Digestion and bioavailability of bioactive phytochemicals

Summary There are many scientific reports on determination of the content and biological activity of compounds found in food. However, these analyses are not sufficient to determine their effect on the human body. During digestion of food ingredients, many changes can modify their structure and this may affect their absorption and bioactivity. Many phenolic aglycones are hydrophilic and can be absorbed through biological membranes by diffusion. However, most polyphenols occur in the glycosidic form, which undoubtedly affects their absorption in the intestine. Oligopeptides are also absorbable via secondary active transport but based on the hydrogen ion gradient or with transporter PepT1. The bioavailability of phytochemicals is determined by their molecular weight or chemical structure and the food matrix. Accordingly, the aim of this work was to present the novel scientific reports related to the influence the many factors on digestibility, bioaccessibility and activity of selected bioactive compounds of plant origin.

Ozone-induced changes in the content of bioactive compounds and enzyme activity during storage of pepper fruits.

This paper presents for the first time the results of investigations concerning the effect of treatment of whole pepper fruits with gaseous ozone and the refrigeration storage period conditions on pepper quality. The effects are reflected in changes in the flavonoid contents, the antioxidant activity of the phenolic compound fraction and the enzymes involved in phenylpropanoid metabolism. The investigations were carried out on a hot pepper fruit cultivar, Cyklon. It was found that the levels of a majority of flavonoids, in particular those of quercetin 3-O-rhamnoside and quercetin 3-O-rhamnoside-7-O-glucoside increased in the pericarp of fruits treated with ozone for 3h and stored for 20days (by 25% relative to the control). Simultaneously, reduced phenylalanine ammonia-lyase and tyrosine ammonia-lyase activity were noted, which implies slight degradation of enzymes caused by the ozone treatment and enhancement of the polyphenol oxidase and guaiacol oxidase activity involved in response to increased oxidative stress.

Identification of potential inhibitory peptides of enzymes involved in the metabolic syndrome obtained by simulated gastrointestinal digestion of fermented bean (Phaseolus vulgaris L.) seeds

The aim of this study was to determine the conditions of bean fermentation carried out by Lactobacillus plantarum to obtain biologically active peptide fractions after in vitro digestion. The results suggest that optimum process conditions should be selected according to the specific activity of peptides. Only a fraction with a molecular mass of 3.5-7kDa obtained after fermentation at 22°C for 3h had α-amylase inhibitory activity. The optimal fermentation conditions for bean seeds to release peptide fractions with a molecular mass of 3.5-7.0kDa and the highest lipase or ACE inhibitory activity (IC50 1.19 and 0.28mg mL-1, respectively) were determined as 30°C and 3days. The fractions with the highest inhibitory activity were identified by LC-MS/MS and the sequences of the peptide derived from bean proteins were determined as INEGSLLLPH, FVVAEQAGNEEGFE, SGGGGGGVAGAATASR, GSGGGGGGGFGGPRR, INEGSLLLPH, GGYQGGGYGGNSGGGYGNRG, GGSGGGGGSSSGRRP, and GDTVTVEFDTFLSR.

ANTIOXIDANT ACTIVITY OF PROTEIN HYDROLYSATES FROM RAW AND HEAT-TREATED YELLOW STRING BEANS (PHASEOLUS VULGARIS L.)

Nowadays, legume plants have been considered not only a source of valuable proteins necessary for the proper functioning and growth of the body but also a source of bioactive compounds such as bioactive peptides, that may be beneficial to human health and protect against negative change in food. The aim of this study was to investigate the effect of heat treatment on the release of antioxidant peptides obtained by hydrolysis of the yellow string beans protein. The antioxidant properties of the hydrolysates were evaluated through free radical scavenging activities (DPPH and ABTS) and inhibition of iron activities (chelation of Fe2+). The results show that the heat treatment had influence on both increased peptides content and antioxidant activity after pepsin hydrolysis of string bean protein. The peptides content after protein hydrolysis derived from raw and heat treated beans were noted 2.10 and 2.50 mg·ml-1, respectively. The hydrolysates obtained from raw (PHR) and heat treated (PHT) beans showed better antioxidant properties than protein isolates (PIR and PIT). Moreover, the hydrolysates obtained from heat treated beans showed the higher ability to scavenge DPPH• (46.12%) and ABTS+• (92.32%) than obtained from raw beans (38.02% and 88.24%, correspondingly). The IC50 value for Fe2+ chelating ability for pepsin hydrolysates obtained from raw and heat treatment beans were noted 0.81 and 0.19 mg·ml-1, respectively. In conclusion, the results of this study showed that the heat treatment string beans caused increase in the antioxidant activities of peptide-rich hydrolysates.