Simultaneous quantification of the degree of hydrolysis, protein conversion rate and mean molar weight of peptides released in the course of enzymatic proteolysis

Beaubier, Sophie; Framboisier, Xavier; Ioannou, Irina; Galet, Olivier; Kapel, Romain

doi:10.1016/j.jchromb.2018.12.005

Cited by 36 publications

(27 citation statements)

References 27 publications

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“…The hydrolysis kinetics were monitored by SEC analyses according to the method developed by Beaubier et al [ 13 ]. A Superdex peptide 10/300 GL column (fractionation range; 10 × 300 mm, GE Healthcare, USA) was used.…”

Section: Methodsmentioning

confidence: 99%

“…The obtained hydrolysates from the SPI were also characterized according to the same method which allows quantifying simultaneously: the DH, the protein conversion rate (Xp) and the mean number of amino acids by peptide produced (Naa), [ 13 ]. The hydrolysate chromatograms were exported in Excel spreadsheets to determine the parameters.…”

Section: Methodsmentioning

confidence: 99%

“…The hydrolysate composition is characterized with two main parameters: the protein conversion rate (Xp) and the mean size of the peptides produced (Naa). The Naa can be expressed by a mean number of amino acids composing the peptide of the hydrolysate and the Xp represents the ratio of protein hydrolyzed on the initial protein at a given time of the hydrolysis reaction [ 13 ]. The advancement of the hydrolysis reaction is commonly determined with the degree of hydrolysis (DH) which represents the ratio of peptide bonds cleaved on protein peptide bonds expressed in percentage [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

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A Rational Approach for the Production of Highly Soluble and Functional Sunflower Protein Hydrolysates

Beaubier

Albe-Slabi

Aymes

et al. 2021

Foods

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Exploitation of plant proteins as an alternative to animal proteins currently presents an important challenge for food industries. In this contribution, total sunflower protein isolate from cold press meal was used as a starting material for the generation of highly soluble and functional hydrolysates that could be used in various food formulations. To do this, a rational and complete approach of controlled hydrolysis was implemented using the individual Alcalase and Prolyve enzymes. The method of stopping the hydrolysis reaction was also evaluated. The influence of operating conditions on hydrolysis kinetics and enzymatic mechanism was studied to identify the appropriate hydrolysis conditions. The gain of the solubility was then analyzed and compared to that of the initial proteins. Finally, the emulsifying and foaming properties (capacities and stabilities) of the resulting hydrolysates were also assessed. As a result, controlled enzymatic proteolysis significantly improved the sunflower protein solubility at neutral pH (twofold increase) and generated highly soluble hydrolysates. The limited proteolysis also maintained the good foam capacities and allowed an improvement in the initial foam stabilities and emulsifying capacities and stabilities of sunflower proteins. This contribution can greatly increase the value of sunflower meal and help in the development of sunflower protein products in the future.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Rational Approach for the Production of Highly Soluble and Functional Sunflower Protein Hydrolysates

Beaubier

Albe-Slabi

Aymes

et al. 2021

Foods

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“…On the other hand, the pH stat technique is simpler and a more direct assay, as it is based on the principle of pH titration. The quantification of the number of terminal end groups liberated from the hydrolysis is calculated based on the amount of NaOH or HCl consumed into the reaction mixture to keep the pH constant during enzymatic hydrolysis [ 91 , 92 , 93 , 94 ]. The reliability of the pH stat technique is in a mathematical relationship between the consumption of the acid/base and the number of the terminal end groups being analyzed.…”

Section: High-resolution Ultrasonic Spectroscopy (Hr-us)mentioning

confidence: 99%

“…Other analytical methods that provide other descriptors of the extent reactions include reverse-phase high-performance liquid chromatography (RP-HPLC) and Fourier-transform infrared spectroscopy (FTIR). Both methods have been utilized to concomitantly determine the degree of hydrolysis, protein conversion rate and molecular weight distribution and explore their complementary natures [ 94 , 95 , 96 , 97 , 98 ]. Both methods are invasive, time-consuming and tedious in sample preparations and data collection and interpretation.…”

Section: High-resolution Ultrasonic Spectroscopy (Hr-us)mentioning

confidence: 99%

Advances in Analysis of Milk Proteases Activity at Surfaces and in a Volume by Acoustic Methods

Dizon

Tatarko

Hianik

2020

Sensors

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This review is focused on the application of surface and volume-sensitive acoustic methods for the detection of milk proteases such as trypsin and plasmin. While trypsin is an important protein of human milk, plasmin is a protease that plays an important role in the quality of bovine, sheep and goat milks. The increased activity of plasmin can cause an extensive cleavage of β-casein and, thus, affect the milk gelation and taste. The basic principles of surface-sensitive acoustic methods, as well as high-resolution ultrasonic spectroscopy (HR-US), are presented. The current state-of-the-art examples of the application of acoustic sensors for protease detection in real time are discussed. The application of the HR-US method for studying the kinetics of the enzyme reaction is demonstrated. The sensitivity of the acoustics biosensors and HR-US methods for protease detection are compared.

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High Metal Chelating Properties from Rapeseed Meal Proteins to Counteract Lipid Oxidation in Foods: Controlled Proteolysis and Characterization

Durand

Beaubier²,

Fine

et al. 2021

Euro J Lipid Sci & Tech

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Rapeseed meal proteins (RP) are enzymatically hydrolyzed using three individual proteases (Alcalase, Flavourzyme, and Prolyve) and the enzymatic mechanism is studied. Rapeseed hydrolysates are produced under controlled conditions and the Prolyve hydrolysate is separated by membrane filtration. Their capacity to reduce free radicals (by transfer of hydrogen or electron) or transition metals (by electron transfer) in the absence of an oxidizable substrate, their metal chelating capacity as well as the antioxidant performances in model (conjugated autoxidizable triene assay) are investigated. All hydrolysates show a reduction capacity (by transfer of hydrogen or electron) and antioxidant activities, in a dose‐dependent manner, which are however not significantly increased in comparison to the native proteins. A noteworthy metal chelating activity of the peptides produced with Prolyve is highlighted. These results indicate the potential of valorization of RP as a source of high metal chelating peptides to counteract lipid oxidation in foods. Practical applications: Over the last decade, the antioxidative potential of peptides from plant biomass has been evidenced by much research. Considering the myriad of possible sources and the diversity of technology and means to obtain peptides from protein materials, it is reasonable to expect more applications. Concomitantly, preventing lipid oxidation, especially with the polyunsaturated fat‐based products, is a major concern in sectors such as agri‐food and cosmetic. Although the efficacy of synthetic antioxidants is recognized, both consumers and manufacturers are looking for more innovative, healthy, environmental‐friendly processes and quality products. In this context, a controlled proteolysis of proteins from plant by‐products can be used as a sustainable strategy to produce antioxidant peptides. Among them, new peptides released from rapeseed proteins with Prolyve can provide interesting metal chelators to counteract lipid oxidation in foods.

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Simultaneous quantification of the degree of hydrolysis, protein conversion rate and mean molar weight of peptides released in the course of enzymatic proteolysis

Cited by 36 publications

References 27 publications

A Rational Approach for the Production of Highly Soluble and Functional Sunflower Protein Hydrolysates

A Rational Approach for the Production of Highly Soluble and Functional Sunflower Protein Hydrolysates

Advances in Analysis of Milk Proteases Activity at Surfaces and in a Volume by Acoustic Methods

High Metal Chelating Properties from Rapeseed Meal Proteins to Counteract Lipid Oxidation in Foods: Controlled Proteolysis and Characterization

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