Production of Brassica carinata Protein Hydrolyzates with a High Fischer's Ratio Using Immobilized Proteases

Abstract

Brassica carinata protein isolates were hydrolyzed using the digestive enzymes trypsin, chymotrypsin, and carboxypeptidase A in order to obtain hydrolyzates with a high Fischer's ratio. The proteases were immobilized using two glyoxyl-agarose supports of different porosity, 4 and 10% agarose gels, in order to evaluate the effect of substrate diffusion into the support containing the enzyme on the hydrolytic process. Reaction time, substrate concentration, and the enzyme to substrate ratio were optimized in an attempt to increase the Fischer's ratio in the resulting hydrolyzates. Gel filtration chromatography of a hydrolyzate with a degree of hydrolysis of 36% yielded a fraction that represented 31% of the total hydrolyzed proteins and had a Fischer's ratio of 28.3 with a phenylalanine + tyrosine content below 1.5%. This material could be used for preparing special diets when there is a need to increase the supply of branched amino acids and/or reduce the intake of aromatic amino acids.

High Fischer ratio oligopeptides determination from Antartic krill: Preparation, peptides profiles, and in vitro antioxidant activity

“…Moreover, the Fischer ratio of PH‐I after removing AAA was 21.12, which was 9.51‐fold of the Fischer ratio of PH‐I before removing AAA. Compared with the literature, HFO with Fischer ratios of 24.58, 32.08, 30.6, and 28.3 from pearl oyster meat protein (Zheng et al, ), tuna meat (Du et al, ; Luo et al, ), bovine casein (Pedroche et al, ), and B. carinata protein (Pedroche et al, ), respectively, were produced through the procedure of protease hydrolysis, activated carbon absorption, and gel permeation chromatography separation. In terms of HFO produced from Antarctic krill as compared to other sources of proteins already investigated, the technology of HFP is a low cost production since the expensive protocol of gel permeation not needed.…”

“…In addition, peptides in HFO usually contain 3-9 amino acids residues (Wei, Huang, Luo, & Song, 2014). At present, HFO prepared by enzymatic hydrolysis of food proteins have been successfully produced from flaxseed (Udenigwe & Aluko, 2010), pearl oyster (Pinctada martensii) (Zheng, Zhang, Cao, Liu, & Ji, 2009), casein (Pedroche et al, 2004), Brassica carinata (Pedroche et al, 2006), corn (Ma et al, 2008;Tanabe et al, 1991), and tuna (Luo, Du, Zheng, Song, & Yang, 2013). HFO from flaxseed proteins could scavenge radical, protect linoleic acid from oxidation and show potential antihypertensive properties by inhibiting angiotensin I-converting enzyme in a mixedtype inhibition pattern (Udenigwe & Aluko, 2010).…”

Obtaining of Brassica carinata protein hydrolysates enriched in bioactive peptides using immobilized digestive proteases

“…Hydrolysis of B. carinata protein isolate was carried out according to the procedure described by Pedroche et al (2006). Protein isolates, 0.4% (w/v) in 50 mM NH 4 HCO 3 , were taken to pH 8.0 and hydrolyzed at 50°C in a batch reactor by addition of the immobilized enzymes in the following order: trypsin, chymotrypsin, and carboxypeptidase A.…”

Protein hydrolysis by immobilized and stabilized trypsin

“…Some years ago, our group was able to prepare stable trypsin catalysts that were useful for the hydrolysis of proteins. 7, 8 These derivatives were obtained by multipoint covalent immobilization (at pH 10) between the region richest in lysines on the trypsin surface and the highly activated glyoxyl supports.…”