Protein Hydrolysis and Glycosylation as Strategies to Produce Bioactive Ingredients from Unmarketable Prawns

The present work shows a procedure to valorize non-commercial boiled shrimp to produce functional ingredients, using a combined treatment based on enzymatic hydrolysis and subsequent glycation under mild conditions. Antioxidant and prolyl endopeptidase-inhibiting activities were determined as a function of hydrolysis and glycation times (0–120 min and 0–180 min, respectively). The reaction products were characterized by determining the degree of hydrolysis, browning, fluorescent compounds, free amino acids, phenol content, Fourier transform infrared spectroscopy (FTIR), and molecular weight of the different fractions obtained. Enzymatic hydrolysis generated hydrolysates with significant antioxidant and prolyl endopeptidase-inhibiting activities. Glycation under mild conditions was used as a strategy to improve the antioxidant and potential nootropic properties of the hydrolysates. During glycation, the free amino acid content decreased, total phenols and fluorescent compounds increased significantly, and low molecular weight melanoidins were formed. The presence of peptide-glucose conjugates was also confirmed by FTIR. Glycation increased the antioxidant activities of the hydrolysates; however, their prolyl-endopeptidase-inhibiting activity was lost. Results showed that compounds with promising antioxidant (hydrolysis and glycation) and potential nootropic (hydrolysis) activities and applications in food systems were obtained from the biotechnological strategy used.

Download Full-text

The effect of enzymatic hydrolysis on the antioxidant activities and amino acid profiles of defatted chia (Salvia hispanica L.) flour

Food Research ◽

10.26656/fr.2017.4(s4).003 ◽

2020 ◽

Vol 4 (S4) ◽

pp. 38-50

Author(s):

E.S.K. Ibrahim ◽

M.A. Ghani

Keyword(s):

Amino Acid ◽

Enzymatic Hydrolysis ◽

Aromatic Amino Acid ◽

Antioxidant Activities ◽

Block Design ◽

Protein Solubility ◽

Amino Acid Content ◽

Degree Of Hydrolysis ◽

Hydrolysis Time ◽

Amino Acid Profiles

The aim of this study was to determine the effect of enzymatic hydrolysis using different proteases (Alcalase® and papain) and hydrolysis period on antioxidative activities and amino acid profiles of hydrolysed chia peptides. The experiment has been carried out using a completely randomized block design. The protein from defatted chia flour (DCF) was first isolated using different extraction pH (pH 10; 11; 12) and precipitation pH (pH 3.5, 4.0 and 4.5) to determine the highest protein isolated (CI) yield. The chia isolate (CI) extracted using the combination treatment (pH 12, 3.5) demonstrated the highest protein content of 17.22% and was selected to further hydrolysed using Alcalase® and papain enzyme at different hydrolysis time. The degree of hydrolysis (DH), protein solubility and peptide content of the chia protein hydrolysates (CH) were observed. Alcalase®-CH and Papain-CH demonstrated the highest DH at 60 mins of hydrolysis with the value of 47.09% and 44.29%, respectively. The protein solubility and peptide content were directly proportional to the DH. The Alcalase®-CH hydrolysed at 60 mins exhibited the highest antioxidant activities as measured by DPPH, ABTS and FRAP assays with values of 35.46µM AAE, 34.45µM TE and 23.11 µM FeSO4.7H2O E, respectively. The Alcalase®- CH demonstrated higher (p<0.05) hydrophobic amino acid (42.51%) compared to and Papain-CH (37.25%,). The highest aromatic amino acid content also recorded by Alcalase®-CH (20.10%), whereas Papain-CH with the value of 15.54%. However, both CH exhibited higher hydrophilic and aromatic amino acid compared to DCF and CI. This result has proved that the enzymatic hydrolysis of CH using Alcalase® and papain improved the nutritional and antioxidant capabilities, thus potentially represent a naturally occurring antioxidant ingredient in the production of functional food and nutraceutical appliance with significant health benefits.

Download Full-text

Effects of Enzymatic Hydrolysis on the Functional Properties, Antioxidant Activity and Protein Structure of Black Soldier Fly (Hermetia illucens) Protein

Insects ◽

10.3390/insects11120876 ◽

2020 ◽

Vol 11 (12) ◽

pp. 876

Author(s):

Inayat Batish ◽

Devon Brits ◽

Pedro Valencia ◽

Caio Miyai ◽

Shamil Rafeeq ◽

...

Keyword(s):

Antioxidant Activity ◽

Protein Structure ◽

Amino Acid ◽

Enzymatic Hydrolysis ◽

Functional Properties ◽

Amino Acid Content ◽

Protein Hydrolysates ◽

Degree Of Hydrolysis ◽

Protein Concentrate ◽

Black Soldier Fly

The effects of chemical protein extraction, and enzymatic hydrolysis with Alcalase, papain and pepsin, on the functional properties, antioxidant activity, amino acid composition and protein structure of black soldier fly (H. illucens) larval protein were examined. Alcalase hydrolysates had the highest degree of hydrolysis (p < 0.05), with the highest hydrolysate and oil fraction yield (p < 0.05). Pepsin hydrolysates showed the lowest oil holding capacity (p < 0.05), whereas no significant differences were observed among other enzymes and protein concentrates (p > 0.05). The emulsifying stability and foam capacity were significantly lower in protein hydrolysates than protein concentrate (p < 0.05). The antioxidant activity of protein hydrolysates from protein concentrate and Alcalase was higher than that with papain and pepsin (p < 0.05), owing to the higher hydrophobic amino acid content. Raman spectroscopy indicated structural changes in protein α-helices and β-sheets after enzymatic hydrolysis.

Download Full-text

Effects of Enzymatic Hydrolysis Conditions on the Antioxidant Activity of Red Tilapia (Oreochromis spp.) Viscera Hydrolysates

Current Pharmaceutical Biotechnology ◽

10.2174/1389201021666200506072526 ◽

2020 ◽

Vol 21 (12) ◽

pp. 1249-1258

Author(s):

Cindy T. Sepúlveda ◽

José E. Zapata

Keyword(s):

Molecular Weight ◽

Antioxidant Activity ◽

Enzymatic Hydrolysis ◽

Protein Concentration ◽

Polynomial Equation ◽

Degree Of Hydrolysis ◽

Red Tilapia ◽

Hydrolysis Conditions ◽

Hydrolysis Of ◽

By Products

Background: Fish is an essential source of nutrients for human nutrition due to the composition of proteins, vitamins, and minerals, among other nutrients. Enzymatic hydrolysis represents an alternative for the use of by-products of the aquaculture industry. Objective: We propose to evaluate the effect of stirring speed, temperature, and initial protein concentration on the degree of hydrolysis of proteins and antioxidant activity of red tilapia (Oreochromis spp.) viscera hydrolysates. Methods: The effect of stirring speed, temperature, and initial protein concentration on the degree of hydrolysis of proteins and antioxidant activity was evaluated using an experimental design that was adjusted to a polynomial equation. The hydrolysate was fractioned to determine the antioxidant activity of the fractions, and functional properties were also measured. Results: Stirring speed and protein concentration presented a statistically significant effect (p <0.05) on all the response variables. However, the temperature did not present a statistically significant effect on the degree of hydrolysis. Discussion: The best conditions of hydrolysis were stirring speed of 51.44 rpm, a temperature of 59.15°C, and the protein concentration of 10 g L-1. The solubility of the hydrolysate protein was high at different pH, and the hydrolysate fraction with the highest antioxidant activity has a molecular weight <1 kDa. Conclusion: The degree of hydrolysis and the biological activity of red tilapia viscera hydrolysates (Oreochromis spp.) are affected by temperature, substrate concentration, and stirring speed. The optimal conditions of hydrolysis allowed to obtain a hydrolysate with antioxidant activity are due to the peptides with low molecular weight.

Download Full-text

Optimization of the Enzymatic Hydrolysis of Tilapia Frames

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.554-556.1387 ◽

2012 ◽

Vol 554-556 ◽

pp. 1387-1394

Author(s):

He Jian Xiong ◽

Longfei Cao ◽

Huajun You ◽

Qingpi Yan ◽

Ying Ma

Keyword(s):

Amino Acids ◽

Enzymatic Hydrolysis ◽

Free Amino Acids ◽

Free Amino ◽

Protein Supplementation ◽

Degree Of Hydrolysis ◽

Enzyme Substrate ◽

Initial Ph ◽

Substrate Ratio ◽

Hydrolysis Of

Tilapia frames were subjected to enzymatic hydrolysis using Flavouzryme and Papain with a ratio of 2:1. The relationship of temperature (40 to 60°C), enzyme: substrate ratio (0.5% to 4.5%), initial pH (6.0 to 8.0) and hydrolysis time (1h to 9h) to the degree of hydrolysis were determined. The enzymatic hydrolysis was optimized for maximum degree of hydrolysis using surface response methodology. The optimum conditions for enzymatic hydrolysis of tilapia frames were temperature 53°C, enzyme : substrate ratio of 3.5%, initial pH 7.2, and reaction time 7h. Under these conditions a degree of hydrolysis of 40.01% were obtained. The yield of free amino acids in the hydrolysate was 46.61mg/g tilapia frames. The flavor amino acids and essential amino acids occupied up to 31.8% and 49.0% of the total free amino acids respectively. The hydrolysate of waste tilapia frames showed good potential for applications such as protein supplementation in food system.

Download Full-text

Anti-Oxidative and Anti-Aging Activities of Porcine By-Product Collagen Hydrolysates Produced by Commercial Proteases: Effect of Hydrolysis and Ultrafiltration

Molecules ◽

10.3390/molecules24061104 ◽

2019 ◽

Vol 24 (6) ◽

pp. 1104 ◽

Cited By ~ 6

Author(s):

Geun-Pyo Hong ◽

Sang-Gi Min ◽

Yeon-Ji Jo

Keyword(s):

Molecular Weight ◽

Antioxidant Activities ◽

Protein Recovery ◽

Degree Of Hydrolysis ◽

Porcine Skin ◽

Amino Group Content ◽

High Antioxidant Activity ◽

Collagen Hydrolysates ◽

Collagenase Inhibition ◽

By Products

To investigate methods for improving the processing of porcine waste, porcine skin was hydrolyzed using different commercially available proteases (Alcalase, Flavorzyme, Neutrase, Bromeline, Protamex, and Papain) under several optimal conditions. Following enzymatic hydrolysis, the collagen hydrolysates (CHs) were fractionated by molecular weight (3 kDa) via membrane ultrafiltration. The CHs were analyzed for physical properties (pH, protein recovery, free amino group content, molecular weight distribution, and amino composition) as well as for functional properties (antioxidant activities and anti-aging activities). Among the CHs, CHs hydrolyzed by Alcalase (CH-Alcalase) exhibited the highest degree of hydrolysis compared to other CHs. Both “CH-Alcalase” and “CH-Alcalase < 3 kDa” fractions showed a considerably high antioxidant activity and collagenase inhibition activity. Therefore, resulting bioactives have potential for development as antioxidants and anti-aging ingredients in the food, cosmetics, and pharmaceuticals, from animal by-products.

Download Full-text

Preparation of Antioxidant Hydrolysates from Round Scad (Decapterus maruadsi) Protein by Composite Enzymatic Hydrolysis

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.941-944.1069 ◽

2014 ◽

Vol 941-944 ◽

pp. 1069-1074 ◽

Cited By ~ 1

Author(s):

Xian Qing Yang ◽

Ting Ting Wang ◽

Xiao Hu ◽

Lai Hao Li ◽

Yan Yan Wu ◽

...

Keyword(s):

Enzymatic Hydrolysis ◽

Incubation Temperature ◽

Antioxidant Activities ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Reducing Power ◽

Degree Of Hydrolysis ◽

Optimum Conditions ◽

High Antioxidant Activity ◽

Substrate Ratio

Round scad (Decapterus maruadsi) was hydrolyzed by composite enzymatic hydrolysis to obtain the hydrolysates possessing high antioxidant activity. The optimum conditions of antioxidant hydrolysates preparation by composite enzymatic hydrolysis were investigated. The results demonstrated that superior antioxidant activities (low IC50 values of radical scavenging activity and high reducing power) were obtained at an enzyme (a mixture of neutral protease and trypsin, 1:1 w/w) to substrate ratio of 0.3%, an incubation time of 5 h and an incubation temperature of 50 °C. Under the conditions, the recovery ratio of protein was 75.24% and the degree of hydrolysis was 25.86%

Download Full-text

Steam Explosion-Assisted Extraction of Protein from Fish Backbones and Effect of Enzymatic Hydrolysis on the Extracts

Foods ◽

10.3390/foods10081942 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1942

Author(s):

Ye Dong ◽

Wen Yan ◽

Xiao-Di Zhang ◽

Zhi-Yuan Dai ◽

Yi-Qi Zhang

Keyword(s):

Enzymatic Hydrolysis ◽

Free Amino ◽

Steam Explosion ◽

Sustainable Use ◽

Fish Bone ◽

Native Fish ◽

Degree Of Hydrolysis ◽

Umami Taste ◽

Assisted Extraction ◽

Pressure Extraction

The development of an efficient pretreatment, prior to enzymatic hydrolysis, is a good strategy for the sustainable use of refractory fish byproducts. This study compared hydrothermal pretreatments at 159 °C for 2 min, followed by water extraction (steam explosion-assisted extraction, SE) and 121 °C for 70 min (hot-pressure extraction, HPE), for the recovery of proteins from fish backbones. The effect of enzymatic hydrolysis on the properties of the obtained fish bone protein (FBP) was also evaluated. The results demonstrated that FBP had high contents of protein (81.09–84.88 g/100 g) and hydroxyproline (70–82 residues/1000 residues). After hydrolysis with Flavourzyme, for 3 h, the FBP hydrolysates that were pretreated with SE (SFBP-H) exhibited a better degree of hydrolysis (DH) and nitrogen recovery (NR), and a higher level of umami taste free amino acids (151.50 mg/100 mL), compared with the HPE-treated samples. The obtained SFBP-H mainly distributed below 3000 Da and had strong scavenging effects on 1,1-diphenyl-2-picrylhydrazy (DPPH) (IC50 = 4.24 mg/mL) and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) (IC50 = 1.93 mg/mL) radicals. Steam explosion-assisted extraction is a promising route for recovering proteins from native fish bone materials, and improving the flavor and antioxidant activity of the hydrolysates.

Download Full-text

Golden Sea Cucumber: Identification and the Antioxidant Activity of Its Collagen Hydrolysates

SQUALEN Bulletin of Marine and Fisheries Postharvest and Biotechnology ◽

10.15578/squalen.v15i3.511 ◽

2020 ◽

Vol 15 (3) ◽

pp. 119

Author(s):

Ir. Yusro Nuri Fawzya, MSi ◽

Nugrah Analiadi Putra ◽

Arif Budi Witarto ◽

Gintung Patantis

Keyword(s):

Molecular Weight ◽

Antioxidant Activity ◽

Sea Cucumber ◽

Antioxidant Activities ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Degree Of Hydrolysis ◽

Dpph Method ◽

Collagen Hydrolysates ◽

Stichopus Horrens

Golden sea cucumber or locally known as “teripang emas” is one of Indonesia’s most popular sea cucumber and widely processed as functional food or supplement due to its bioactivities. The sea cucumber is often misidentified due to its morphological similarities with other Stichopus spp. This study aimed to identify the golden sea cucumber obtained from West Nusa Tenggara, Indonesia, by a molecular method and study the antioxidant activities of its collagen hydrolysates. The hydrolysates were produced by hydrolyzing acid collagen extract using neutrase for 30, 60, 120, 180, and 240 mins. The products were then analyzed for their degree of hydrolysis, peptide content, molecular weight distribution and radical scavenging activity by the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) method. Results showed that hydrolysis for 180 mins was optimal in producing the highest peptide content, 12.79 ± 0.44 mg/mL, with a degree of hydrolysis (DH) of 55.2 ± 1.50%. However, the highest antioxidant activity (IC50 of 5.25 ± 0.15 mg/mL) was demonstrated after 60 mins hydrolysis with molecular weight (MW) ranged from less than 14.4 kDa to approximately 25 kDa. The hydrolysate might be categorized as a weak to moderate antioxidant. Based on the molecular identification, the golden sea cucumber had 99% similarities with Stichopus horrens and S. monotuberculatus.

Download Full-text

Antioxidant and chelating activities from Lion fish (Pterois volitans L.) muscle protein hydrolysates produced by in vitro digestion using pepsin and pancreatin.

Emirates Journal of Food and Agriculture ◽

10.9755/ejfa.2020.v32.i1.2060 ◽

2020 ◽

pp. 62

Author(s):

Luis Chel-Guerrero ◽

David Cua-Aguayo ◽

David Betancur-Ancona ◽

Azucena Chuc-Koyoc ◽

Irma Aranda-González ◽

...

Keyword(s):

Molecular Weight ◽

Amino Acid ◽

Acid Content ◽

Amino Acid Content ◽

Fish Muscle ◽

Degree Of Hydrolysis ◽

Chelating Activity ◽

Freeze Dried ◽

Sds Page

The aim of the present research was to determine the antioxidant, and Cu2+ and Fe2+ chelating activities, amino acid content and estimated molecular weight of proteins and peptides from lion fish muscle-derived hydrolysates obtained by enzymatic hydrolysis with pepsin and pancreatin. The fillets were freeze-dried and hydrolysis was performed, taking aliquots at different times. The degree of hydrolysis, antioxidant activity against ABTS+ and DPPH, Cu2+ and Fe2+ chelating activity, as well as molecular weight estimated by SDS-PAGE and amino acid content by HPLC was determined. The highest degree of hydrolysis (DH) was at 180 min (37.75%). SDS-PAGE showed proteins with an estimated molecular weight of 45,259 and 42,487 Da, which could be associated with myofibrillar proteins. The progressive degradation of proteins by enzymes was also observed, detecting polypeptides with a EMW of 5,988 and 4,954 Da in the most representative hydrolysates. The H40 hydrolysate exhibited the highest ABTS and DPPH radical depuration activity, with values of 107.31 mM/mg protein and 54.27%, respectively. The iron and copper chelating activity was related to DH, since the highest values of iron and copper chelating activity were obtained in hydrolysates H120 and H140, with 90.83% chelation of Cu2+ and 56.33% chelation of Fe2+, respectively with no significant differences compared to subsequent times. In addition, the antioxidant and chelating activities were possibly related to Trp, Cys, Lys, Pro and Arg content. Lion fish muscle hydrolysates could be a potential source of functional ingredients due to their in vitro antioxidant and chelating activity.

Download Full-text