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Marquez-Rios, Enrique,Cota-Arriola, Octavio,Villalba-Villalba, Ana Gloria,Ezquerra-Brauer, Josafat Marina,Ocano-Higuera, Victor Manuel,Lopez-Corona, Betzabe Ebenhezer,Torres-Arreola, Wilfrido 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.4
Chymotrypsin was purified from jumbo squid hepatopancreas (HP) with 2.4-fold and yield 1.9%, and characterized with a molecular weight of 31 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Chymotrypsin effect over collagen extracted from the mantle, fins and arms of the jumbo squid was evaluated. The enzyme exhibited the maximum activity at pH 7 and $65^{\circ}C$ using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as a substrate and it was identified using the specific inhibitors N-tosyl-L-phenylalaninechloromethyl ketone (TPCK) and phenyl methyl sulfonyl fluoride (PMSF), showing residual activities of 6% and 0%, respectively. Furthermore, high activity was observed in the pH range of 4.0 to 8.0. Purified enzyme showed a moderate in vitro activity using muscle collagen as a substrate. Although further research is needed, the results suggest that the enzyme has a potential application where acidic or slightly alkaline conditions are needed.
Sarabia-Sainz, Hector M.,Ezquerra-Brauer, Josafat Marina,Santacruz-Ortega, Hisila C.,Rouzaud-Sandez, Ofelia,Valenzuela-Soto, Elisa M.,Acosta-Elias, Monica,Torres-Arreola, Wilfrido 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.1
Muscle from mantle, fins and arms of squid (Dosidicus gigas) were compared based on lysyl oxidase activity (LOX), chemical/structural and thermodynamic properties of highly cross-linked collagen. The arms collagen presented the highest temperature (Tp) and enthalpy of transition. The arms collagen thermic properties may be explained by the higher imino amino acid content, proline and lysine hydroxylation degrees. Moreover, among the regions, the collagen from the arms had a more intense ${\beta}$ band chain, hydroxymerodesmosine peak in the resonance magnetic nuclear spectra and pyridinoline peak in the Raman spectra. Fins showed the highest LOX activity. The LOX activity was associated with the Tp, proline and lysine hydroxylation degrees. These results implied that the collagen in the arms was more intermolecularly ordered than the mantle and fins, and may provide a theoretical basis for a better understanding of the thermal behaviour of squid tissues during management and processing.
Enrique Marquez-Rios,Octavio Cota-Arriola,Ana Gloria Villalba-Villalba,Josafat Marina Ezquerra-Brauer,Victor Manuel Ocaño-Higuera,Betzabe Ebenhezer Lopez-Corona,Wilfrido Torres-Arreola 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.4
Chymotrypsin was purified from jumbo squid hepatopancreas (HP) with 2.4-fold and yield1.9%, and characterized with a molecular weight of 31 kDa, as estimated by sodium dodecyl sulfatepolyacrylamidegel electrophoresis (SDS-PAGE). Chymotrypsin effect over collagen extracted from themantle, fins and arms of the jumbo squid was evaluated. The enzyme exhibited the maximum activityat pH 7 and 65oC using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as a substrate and it was identifiedusing the specific inhibitors N-tosyl-L-phenylalaninechloromethyl ketone (TPCK) and phenyl methylsulfonyl fluoride (PMSF), showing residual activities of 6% and 0%, respectively. Furthermore, highactivity was observed in the pH range of 4.0 to 8.0. Purified enzyme showed a moderate in vitro activityusing muscle collagen as a substrate. Although further research is needed, the results suggest that theenzyme has a potential application where acidic or slightly alkaline conditions are needed.
He´ctor M. Sarabia-Sainz,Josafat Marina Ezquerra-Brauer,Hisila C. Santacruz-Ortega,Ofelia Rouzaud-Sa´ndez,Elisa M. Valenzuela-Soto,Monica Acosta-Elias,Wilfrido Torres-Arreola 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.1
Muscle from mantle, fins and arms of squid (Dosidicus gigas) were compared based on lysyl oxidase activity (LOX), chemical/structural and thermodynamic properties of highly cross-linked collagen. The arms collagen presented the highest temperature (Tp) and enthalpy of transition. The arms collagen thermic properties may be explained by the higher imino amino acid content, proline and lysine hydroxylation degrees. Moreover, among the regions, the collagen from the arms had a more intense b band chain, hydroxymerodesmosine peak in the resonance magnetic nuclear spectra and pyridinoline peak in the Raman spectra. Fins showed the highest LOX activity. The LOX activity was associated with the Tp, proline and lysine hydroxylation degrees. These results implied that the collagen in the arms was more intermolecularly ordered than the mantle and fins, and may provide a theoretical basis for a better understanding of the thermal behaviour of squid tissues during management and processing.