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Przedmiotem badań była ocena właściwości przeciwutleniajacych hydrolizatów preparatu foswitynowego z jaj kurzych. Hydrolizę prowadzono z udziałem trypsyny wołowej i z proteazy z A. melleus. Wyznaczono stopień hydrolizy (DH), stężenie wolnych grup aminowych i przedstawiono profile RP-HPLC uzyskanych peptydów. W produktach oznaczano: zdolność do wymiatania wolnych rodników DPPH, chelatowania jonów żelaza (II) (FRAP) oraz siłę redukującą. Finalny stopień hydrolizy wyniósł: 33,7 i 23,2% odpowiednio dla proteazy z A. melleus i dla trypsyny. Potwierdzono to analizą przyrostu wolnych grup aminowych, których końcowe stężenie osiągnęło: 3816 μM · g-1 i 1198,5 μM ·g-1 dla proteazy z A. melleus i dla trypsyny. 24-godz. hydrolizat trypsynowy (0,27 μM Trolox · mg-1) wykazał większą zdolność wymiatania wolnych rodników DPPH niż hydrolizat otrzymany proteazą z A. melleus (0,21 μM Trolox · mg-1). Uzyskane hydrolizaty charakteryzowały się bardzo wysoką zdolnością chelatowania jonów Fe (II). Najwyższy poziom tej aktywności wynoszący: 1466,3 μg Fe2+ ·mg-1 uzyskano dla hydrolizatu (24-godz.) otrzyma- nego z zastosowaniem proteazy z A. melleus.
Increasing the potency of antihypertensive food-derived peptides is a critical and important step in the development of natural drugs for cardiovascular diseases prevention. We have proposed the egg-white protein precipitate (EWPP) obtained as a byproduct of cystatin and lysozyme isolation as a potential source of ACE-inhibitory peptides derived by pepsin digestion. The results indicated that hydrolysis of EWPP with pepsin produced the ACE inhibitory activity. During 3-h hydrolysis (DH: 38.3%), the IC50 value of EWPP hydrolysate was signifi cantly increased and fi nally reached IC50=643.1 μg/mL. This hydrolysate was further fractionated by RP-HPLC. The peptide fraction exhibiting the highest ACE inhibitory activity was rechromatographed. Three peptide subfractions exhibiting ACE-inhibitory activities of 69.0, 25.0, and 37.6 μg/mL were further characterised. In each of them, mixtures of peptides with different molecular masses were observed.
Biologically active peptides are of particular interest to food science and nutrition because they may act as potential physiological modulators of metabolism. Hidden or inactive in the amino-acid sequence of food proteins, they can be released or activated in vivo during gastrointestinal digestion, or in vitro during enzymatic hydrolysis and food processing. Egg proteins are an important source of these bioactive peptides. In recent years, major egg protein components, ovoalbumin, conalbumin, ovomucin and phosvitin, have also been shown to contain bioactive sequences. Peptides showing antiadhesive, anticancer as well as immunomodulatory activities were found in ovomucin sequence. Immunoregulative peptides were also found in peptic and chymotryptic hydrolysates of ovoalbumin. Ovocinin and ovocinin(2-7) - angiotensin-converting enzyme inhibitory peptides, whose pharmacological activity was observed in micromolar concentrations - were liberated during in vitro proteolysis of ovoalbumin. Furthermore, many egg protein hydrolysates showed antimicrobial and antioxidant activity after proteolysis with different enzymes, and several active peptides were isolated and identified.
Celem podjętych w pracy badań była ocena podatności wybranych białek serwatkowych na działanie zewnątrzkomórkowych proteaz drożdży Yarrowia lipolyti- ca i określenie ich potencjalnej przydatności technologicznej w procesie enzymatycznej hydrolizy. Do badań wykorzystano zarówno aktywną w środowisku kwaśnym proteazę aspartylową, a także działającą w środowisku alkalicznym proteazę serynową. Hydroli­zie poddano a-laktoalbuminę, P-laktoglobulinę oraz dostępny w handlu koncentrat białek serwatkowych (WPC-80). Reakcję hydrolizy prowadzono w temperaturze 37°C, w pH 3,0 (dla proteazy aspartylowej) i w pH 8,0 (dla proteazy serynowej), wprowadzając enzym w dawce 10 U/mg białka substratowego. Postęp proteolizy analizowano ilościowo poprzez oznaczenie stopnia hydrolizy (DH%) i przyrost wolnych grup aminowych oraz jakościo­wo, tj. elektroforetycznie, a także wykonując rozdział frakcji białkowo-peptydowych przy wykorzystaniu wysoko sprawnej chromatografii cieczowej w układzie odwróconych faz (RP-HPLC). W puli wszystkich przetestowanych wariantów badawczych najwyższy po­ziom degradacji sięgający 39% uzyskano w roztworach P-laktoglobuliny trawionych droż­dżową protezą serynową.
 The main objective of this study was to determine potential application of a serine proteinase derived from Asian pumpkin for obtaining biologically active peptides from casein. The course of casein hydrolysis by three doses of the enzyme (50, 150, 300 U/mg of protein) was monitored for 24 hours by the determinations of: hydrolysis degree DH (%), free amino group content (μmole Gly/g), RP HPLC peptide profiles and by polyacrylamide gel electrophoresis. In all hydrolyzates analyzed antioxidant activities were determined using three tests: the ability to reduce iron ions in FRAP test, the ability to scavenge free radicals in DPPH test, and Fe2+ chelating activity. The antimicrobial activity of obtained peptide fractions was determined as the ability to inhibit the growth of Escherichia coli, Bacillus cereus and Pseudomonas fluorescens in a diffusion plate test. The deepest degradation, expressed as the DH [%] and the free amino group content (67% and 7528 µmole Gly/mg, respectively), was noted in samples hydrolyzed with 300 U/ml of enzyme for 24 hours, while in other samples the determined values were about three and two times lower. The results were in agreement with the peptide profiles obtained by RP HPLC. The highest antioxidative activities determined in all tests were seen for the casein hydrolysate obtained with 300 U/mg protein of serine proteinase after 24 h of reaction (2.15 µM Trolox/mg, 96.15 µg Fe3+/mg, 814.97 µg Fe2+/mg). Antimicrobial activity was presented in three preparations. In other samples no antimicrobial activity was detected.
Natural antioxidants with high safety and long-term effect are subject of many studies because they are an alternative to chemical antioxidants which, in larger quantities, can be toxic. One type of natural antioxidants may be food- derived peptides. The aim of this study was to obtain antioxidant and antimicrobial peptides from egg yolk phosvitin and immunoglobulin Y (IgY) with participation of microbial proteinases from Bacillus amyloliquefaciens (neutrase), B. thermoproteolyticus Rokko (thermolysin), Streptomyces griseus (pronase) and Aspergillus melleus. The progress of hydrolysis was monitored by the degree of hydrolysis (DH) and free amino groups concentration measurement. The resulting hydrolysates were subjected to an assessment of their ability to reduce the oxidation state of metal ions, scavenging of 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) free radicals and chelating iron ions. The highest degree of hydrolysis of both proteins was obtained during the reaction with proteinase from S. griseus. The highest level of reduction in the oxidation state of iron ions was observed in IgY 24-hour hydrolysates obtained with the participation of enzyme from B. thermoproteolyticus Rokko (409.7 μg Fe²+/mg). However, the 24-hour hydrolysates of IgY obtained after degradation with the proteinase from A. melleus, possessed the highest free radical scavenging activity equal to1.46 μM trolox/mg. The highest activity of chelating iron ions, equal to 891.64 μg Fe²+/mg, was observed for products obtained during the 24-hour hydrolysis of phosvitin with the participation of protease from B. thermoproteolyticus Rokko.
A Yarrowia lipolytica JII1c yeast strain, isolated from the Polish ‘Rokpol’ mould cheese, was used as an adjunct culture in the production of a Dutch-type cheese. Its effect on the microbiological and biochemical characteristics of the cheese was evaluated in this research study. Milk used to produce the cheese was inoculated with 105 cfu/mL yeast cells. During the ripening process, the yeast population grew systematically to reach a maximum level of 7.9 log cfu/g in the sixth week of maturation, whereas the number of lactic acid bacteria increased until the fourth week of ripening. Thereafter, the number of microorganisms in the both groups decreased. After 8 weeks of ripening, the pH value of cheese inoculated with yeasts was significantly higher than that of the control cheese sample (produced without those microorganisms) and reached the levels of 6.37 and 5.47, respectively. In the experimental cheeses, it was also found that the utilization rate of lactic and citric acids was higher. Additionally, the concentration levels of water-soluble nitrogen (WSN) and free amino groups (FAG) in the experimental cheeses were about twice as high as in the control cheese sample. A more intensive proteolysis in the experimental cheese was accompanied by a higher accumulation of biogenic amines, especially of tyramine, putrescine, and 2-phenylethylamine; in the experimental cheese, after 8 weeks, their contents amounted to: 167.01, 77.90, and 69.54 mg/100 g, respectively. In contrast, the concentration of histamine was similar in both cheeses (9.47 and 9.81 mg/100 g in the control and experimental cheese samples, respectively). Also, the experimental cheese revealed more pronounced lipolysis resulting in a higher accumulation of free fatty acids, especially of butyric, myristic, palmitic, stearic, and oleic acids. It can be concluded that the Y. lipolytica JII1c grew well in the cheese causing the ripening process of the cheese to significantly accelerate.
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