## Abstract One‐dimensional and two‐dimensional SDS‐PAGE and Hydrophobic Cluster Analysis (HCA) were used for investigating the course of limited tryptic hydrolysis of faba bean legumin. SDS‐PAGE revealed the formation of an “unequal” subunit pattern, characterized by 30 and 50 kDa subunits, which
Changes in interfacial properties of legumin from faba beans (Vicia faba I.) by tryptic hydrolysis
✍ Scribed by Krause, J.-P. ;Schwenke, K. D.
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 589 KB
- Volume
- 39
- Category
- Article
- ISSN
- 0027-769X
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✦ Synopsis
Changes of the surfxe tension at a planar interface and of the emulsifying properties of tryptically digested legumin from faba beans (enzyme/substrate ratio = 1/40) were studied in terms of time of hydrolysis (0-48 h). Simultaneously, molecular changes of legumin were studied by determination of liberated amino groups, RP-HPLC. and SDS-PAGE analysis.
While the surface tension continuously decreases with increasing amount of liberated amino groups, improved emulsion properties, i.e. decreasing droplet size and increasing coalescence as well as creaming stability, were only observed during the first 240 min of hydrolysis, where incredse in the content of free amino groups amounted to 18-24 free amino groups per protein subunit. The surface tension during the diffusion-controlled phase reached the lowest value after a hydrolysis time of 24 h which corresponds to an increase of 44-50 free amino groups per protein subunit. Additionally, the results support the view that the interfacial adsorption at quiet interfaces and during emulsion formation are governed by different transport mechanisms.
Zusammenfassung
Veranderungen des Adsorptionsverhaltens des Legumins der Ackerbohne (Vicia $aha L.) an der planaren Luft/Wasser-Grenzflache sowie der Emulgieraktivitat wurden in Abhangigkeit von der Dauer der tryptischen Hydrolyse (0-48 h) bei einem Enzym/Substratverhlltnis von 1/40 untersucht. Die Bestimmung der Zahl freier Aminogruppen (NH,) sowie RP-HPLC und SDS-PAGE-Untersuchungen gaben AufschluB uber strukturelle Veranderungen des Legumins wahrend der Hydrolyse. Die Oberflachenspannung nimmt nahezu linear mit steigender Zahl freigesetzter Aminogruppen a b und erreicht den niedrigsten Wert nach einer Hydrolysezeit von 24 h (44 -50 Aminogruppen pro Untereinheit). Verbesserungen der Emulsionseigenschaften, wie abnehmende TropfchengroBe oder erhohte Koaleszenz-und Aufrahmstabilitlt, sind nur wiihrend der ersten 240 min der Hydrolyse zu verzeichnen. Die Zahl zusltzlicher freier Aminogruppen lag zwischen 18 und 24 Aminogruppen pro Leguminuntereinheit. Die Ergebnisse stiitzen die These, dal3 unterschiedliche Transportmechanismen und somit verschiedene Fraktionen des hydrolysierten Leguminmolekuls die diffusionskontrollierte Phase der Adsorption und die Homogenisierung bestimmen.
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## Abstract Kinetics and mechanism of proteolysis of broad bean (__Vicia faba__ L.) legumin by trypsin in concentrated solutions at different enzyme‐substrate ratios (__E/S__ = 1/40, 1/100, 1/200, 1/1000) were studied. By the method of HPLC it was established that during proteolysis process (__E/S_