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Production of enzyme hydrolysates from flaxseed proteins and evaluation of their properties
HAŠKOVÁ, Kateřina
This thesis focuses on effect on antioxidant properties and chosen functional properties: solubility, water and fat holding capacity, emulsification activity and stability caused by hydrolysis of protein isolates (PI) made from three different flax varieties (Libra, Agriol and Raciol). Part of this work was production of PI from flax seed. For hydrolysis trypsin, alcalase and papain were used - hydrolysis by trypsin was the mildest, while hydrolysis by papain was the most intensive one. Dynamics and degree of hydrolysis were also observed. Solubility of PI and hydrolysates was discovered to be very high (almost 100%). On the contrary water holding capacity was lower (3,93 g/g and less) than fat holding capacity (4,34 g/g), which increased upon hydrolysis for most enzymes, while water holding capacity decreased. Emulsification activity rised (1,47×) under the effect of hydrolysis, however stability dropped (1,3×). The best emulsifiction properties were found out in trypsin hydrolysates. Hydrolysis caused great impact on content of polyphenols which were in alcalase hydrolysates up to 3× higher and on antioxidant activity which grew in papain hydrolysates up to 2,5×.
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Redox and catalytic properties of modified papain
Lachmanová, Štěpánka ; Hudeček, Jiří (advisor) ; Zima, Jiří (referee)
Redox and Catalytic Properties of Modified Papain Endopeptidase papain could be modified by the organometallic complex [(η6 -C6H5(CH2)2NHCO(CH2)Cl)Ru(N^N)Cl]Cl, (N^N) = 1,10-phenanthroline (TB11), which binds to the free sulfhydryl group of cysteine in the active centre of protein. This modification influences the catalytic properties of papain. The redox properties of the TB11 complex in its free form and when it is bound to papain were studied by the polarographic and voltammetric methods. Modified endopeptidase is reduced in the adsorbed state. Therefore, a phase-sensitive AC voltammetry is used for studies of the electron transfer in the TB11 modified papain. This method gives suitable methodology for studying the electron transfer rates in the systems influenced by adsorption. Using this method the catalytic hydrogen evolution process was detected not only in the TB11, but also for TB11 covalently-attached to the papain molecule. The results help better understand the catalytic mechanism of the hydrogenation reactions in the presence of TB11 modified enzyme. Adsorption properties of papain at different substrates were studied by AFM measurement. The formation of a compact monolayer was proved for the gold substrate modified by a layer of 11-mercaptoundecanoic acid.
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Catalytic and adsorption properties of papain and its derivatives
Lachmanová, Štěpánka ; Hudeček, Jiří (advisor) ; Karpenko, Vladimír (referee)
The aminoacid sequence of papain (EC 3.4.22.2) consists of 212 aminoacids. It has only one free sulfhydryl group, which is located in the active site of the protein. Some organometallic complexes could be bonded only to this free -SH group due to their structure. The artificial metalloproteins synthesised by this way may have different electrochemical properties. In this work, we have studied the electrochemical properties of papain and its derivatives. We compared the ability of papain and its three artificial derivatives to catalyse the hydrogen evolution by the chronopotenciometry. The work was completed by the study of the electrochemical properties of the organometallic complexes of ruthenium, which were used for the artificial metalloprotein preparation. The electrochemical properties of the compounds were never studied before. The process of the hydrogen evolution catalysed by the proteins is held in the adsorbed state of the catalyst. Due to this fact we have also studied the adsorption properties of papain on the substrates with different level of hydrofobicity. (In Czech)
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Chronopotentiometry of Papain Modified by Ruthenium Complexes
Nováková Lachmanová, Štěpánka ; Pospíšil, Lubomír ; Talbi, B. ; Salmain, M. ; Hromadová, Magdaléna
Catalytic hydrogen evolution reaction is one of known effects observed during the electrochemical studies of proteins. Constant current chronopotentiometric stripping technique is suitable tool for the study of catalytic hydrogen evolution reaction due to the formation of peak H. This contribution compares the catalytic behaviour of nonmodified papain and its artificial derivatives prepared by the interaction of organometallic complexes of ruthenium with free sulfhydryl group of protein. The comparison of the chronopotentiometric behaviour of papain and its derivatives would help to better understand the catalytic hydrogen evolution reaction in these derivatives.
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Catalytic and adsorption properties of papain and its derivatives
Lachmanová, Štěpánka ; Hudeček, Jiří (advisor) ; Karpenko, Vladimír (referee)
The aminoacid sequence of papain (EC 3.4.22.2) consists of 212 aminoacids. It has only one free sulfhydryl group, which is located in the active site of the protein. Some organometallic complexes could be bonded only to this free -SH group due to their structure. The artificial metalloproteins synthesised by this way may have different electrochemical properties. In this work, we have studied the electrochemical properties of papain and its derivatives. We compared the ability of papain and its three artificial derivatives to catalyse the hydrogen evolution by the chronopotenciometry. The work was completed by the study of the electrochemical properties of the organometallic complexes of ruthenium, which were used for the artificial metalloprotein preparation. The electrochemical properties of the compounds were never studied before. The process of the hydrogen evolution catalysed by the proteins is held in the adsorbed state of the catalyst. Due to this fact we have also studied the adsorption properties of papain on the substrates with different level of hydrofobicity. (In Czech)
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Redox and catalytic properties of modified papain
Lachmanová, Štěpánka ; Zima, Jiří (referee) ; Hudeček, Jiří (advisor)
Redox and Catalytic Properties of Modified Papain Endopeptidase papain could be modified by the organometallic complex [(η6 -C6H5(CH2)2NHCO(CH2)Cl)Ru(N^N)Cl]Cl, (N^N) = 1,10-phenanthroline (TB11), which binds to the free sulfhydryl group of cysteine in the active centre of protein. This modification influences the catalytic properties of papain. The redox properties of the TB11 complex in its free form and when it is bound to papain were studied by the polarographic and voltammetric methods. Modified endopeptidase is reduced in the adsorbed state. Therefore, a phase-sensitive AC voltammetry is used for studies of the electron transfer in the TB11 modified papain. This method gives suitable methodology for studying the electron transfer rates in the systems influenced by adsorption. Using this method the catalytic hydrogen evolution process was detected not only in the TB11, but also for TB11 covalently-attached to the papain molecule. The results help better understand the catalytic mechanism of the hydrogenation reactions in the presence of TB11 modified enzyme. Adsorption properties of papain at different substrates were studied by AFM measurement. The formation of a compact monolayer was proved for the gold substrate modified by a layer of 11-mercaptoundecanoic acid.
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