Comparison the effect of three commercial enzymes for enzymatic hydrolysis of two substrates (rice bran protein concentrate and soy-been protein) with SDS-PAGE

Abstract

In this research enzymatic hydrolysis of rice bran protein concentrate (RBPC) and soybean Protein (SBP) as control were studied with 3 commercial enzymes (Alcalase®, Papain and acommercial 3-enzyme cocktail containing of 1.6 mg ml − 1 Trypsin, 3.1 mg mlChymotrypsin, 1.3 mg ml −1 Aminopeptidase (SIGMA P7500) and 7.95 mg ml −1 pronase type XIV (SIGMA P5147) by the pH stat method. The hydrolysis was carried out at temperature of 28 C, 60 min and pH 8.00. Results were showed that RBPC, and SBP had higher Degree h y d r o l y s i s ( D H % ) w i t h A l c a l a s e ® e n z y m e . Alcalase®had stronger capability for hydrolysis compared to the other tested enzymes. After 60 minute of hydrolysis time, the DH% of Alcalase® for RBPC and SBP was 12.69 and 12.50 %, respectively. In contrast, papain enzyme was showed lowest DH% in three substrates that 1.56 and 1.24 % were for SBP and RBPC, respectively.The hydrolysis of the protein fraction performed the three enzymes on the two substrates was followed in SDS-PAGE. RBPC and SBP showed almost complete digestion with Alcalase® enzyme after 60 minutes. 3-enzyme cocktail enzyme hydrolyzed better the RBPC than the SBP. Papain enzyme had less effect on the two substrates than other 2 enzymes. It was found that Alcalase® has highest capability for hydrolysis compared to other enzyme preparations. The high value protein hydrolysates prepared by Alcalase® can be used as value added ingredients in many food formulations. They are also suitable for a broad range of industrial food applications and also for cosmetic and personal care products.

Online monitoring of enzymatic hydrolysis of marine by-products using benchtop nuclear magnetic resonance spectroscopy

“…Comparing results, it appears that the reactions monitored via NMR reach equilibrium more quickly than those in the literature monitored by other methods. In particular, while both the NMR measurements here and measurements in the literature (Pagan et al 2015;Ahmadifard et al 2016) showed a similar rapid increase at the beginning of hydrolysis, the results from the NMR plateaued sooner than results measured by other methods.…”

“…The calculated reaction rates approximate rates found in the literature for hydrolysis using Alcalase and Flavourzyme enzymes under similar experimental conditions (Ahmadifard 2016;Sundekilde et al 2018; Kristoffersen et al 2019). It was observed that the combined Alcalase and Flavourzyme measurement on the red cod produced a slower reaction and a less efficient hydrolyzation than for the Alcalase alone.…”

Combinations of Legume Protein Hydrolysates Synergistically Inhibit Biological Markers Associated with Adipogenesis

“…On the other hand, due to the very broad substrate specificity, alcalase can hydrolyze most peptide bonds within a protein molecule. Therefore, its use on substrates like legume proteins causes a high degree of hydrolysis, producing many peptides of small sizes [ 43 ]. The low DH values obtained in fava bean and chickpea LPH with alcalase could be related to vicilin, as this protein is known for its emulsifying, foaming and gelling properties [ 44 , 45 ].…”

Characterization of a soy protein hydrolyzate for the development of a functional ingredient

“…The commercial enzyme used for the hydrolysis showed to be very efficient compared to the enzymes used by Kong et al (2008) with only 18.36 DH % in 180 min. Ahmadifard et al (2016) used the commercial enzyme Alcalase to hydrolyze soy protein, obtaining 12.50 DH % after 60 min, whereas Corolase 7089 reached 47.91 DH % at 60 min. A reasonably low concentration of enzyme was needed to obtain hydrolysates.…”