Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

Acid hydrolysis of gelatin extracted from cow skin: properties and potential for use as a source of small peptides and free amino acids for broiler chickens

K. Nouri A , S. Khalaji https://orcid.org/0000-0002-6518-9647 A D , A. Zamani B and A. Saki https://orcid.org/0000-0002-6789-1288 C
+ Author Affiliations
- Author Affiliations

A Department of Animal Science, Faculty of Agricultural Sciences, Malayer University, Malayer, 65719-95863, Iran.

B Department of Fisheries, Faculty of Natural Resources and Environment, Malayer University, Malayer, 65719-95863, Iran.

C Department of Animal Science, Bu-Ali Sina University, District 2, Hamedan, Iran.

D Corresponding author. Email: saeed.khlj@gmail.com

Animal Production Science - https://doi.org/10.1071/AN20411
Submitted: 11 July 2020  Accepted: 5 October 2020   Published online: 26 October 2020

Abstract

Context: Acid hydrolysis of animal and plant protein ingredients can generate beneficial and cost-effective peptides and free amino acids for use in livestock feed.

Aims: The aim was to determine whether a cow-skin gelatin (CSG) hydrolysate, rich in low-molecular-weight peptides and free amino acid and produced through acid (4 M HCl) hydrolysis, improved the digestibility of CSG and performance of broilers.

Methods: Day-old Ross 308 chicks (n = 320) were allocated to four treatments (control, maize–soy basal diet; CSG at 92 g/kg basal diet; acid-hydrolysed CSG (AHCSG) at 45 and 92 g/kg basal diet) in a completely randomised design with eight replicate pens per treatment and 10 chicks in each pen. Molecular weights of CSG and AHCSG were characterised by SDS–PAGE, and protein concentration and degree of hydrolysis of AHCSG determined. Broilers were assessed for performance measures and intestinal and serum characteristics.

Key results: The AHCSG had a protein concentration of 84.3%, with a degree of hydrolysis of 66.5%. Respective molecular weights of CSG and AHCSG were 20–128 kDa and 3.5–10 kDa. Bodyweight gain and feed intake were dramatically (P ≤ 0.001) reduced and feed conversion ratio increased with inclusion of CSG and especially AHCSG in the diet. Inclusion of AHCSG reduced (P < 0.05) ileal digesta viscosity compared with CSG, and reduced (P < 0.05) plasma uric acid concentration, villi height and crypt depth compared with the CSG and control diets. AHCSG inclusion in the diet reduced protein digestibility by ~25% (at 45 g/kg) and 50% (at 92 g/kg) compared with the control; CSG also reduced protein digestibility by ~50%. The Salmonella population of ceca was reduced (P = 0.05) with the diet containing 92 g AHCSG/kg compared with the CSG and control diets. Trypsin activity was not affected by diet, but total alkaline protease activity was reduced (P < 0.01) at days 35 of age by inclusion of AHCSG in the diet. Carcass and breast relative weight, and breast fillet crude protein were lower (P < 0.05) for broilers fed diets containing CSG or AHCSG than the control.

Conclusions: Acid hydrolysis of CSG produced large quantities of free amino acids and small peptides, but this did not improve the performance of broiler chickens compared with CSG. Performance was similarly poor with dietary inclusion of AHCSG and CSG relative to the control diet without CSG products.

Implications: Although large quantities of free amino acids and small peptides were produced by acid hydrolysis of CSG, further research is needed to understand the possible problems with acid hydrolysis and why it negatively affects broiler performance.

Keywords: acid, alkaline, carcass, digestibility, free amino acids, gelatin, hydrolysis, peptide.


References

Agyei D, Potumarthi R, Danquah MK (2015) Food-derived multifunctional bioactive proteins and peptides. In ‘Biotechnology of bioactive compounds: sources and applications’. (Eds VK Gupta, MG Tuohy) pp. 483–505. (John Wiley & Sons: Chichester, UK)

Aluko R (2012) Bioactive peptides. In ‘Functional foods and nutraceuticals’. pp. 37–61. (Springer: New York)

Álvarez C, Bances M, Rendueles M, Díaz M (2009) Functional properties of isolated porcine blood proteins. International Journal of Food Science & Technology 44, 807–814.
Functional properties of isolated porcine blood proteins.Crossref | GoogleScholarGoogle Scholar |

Álvarez C, Bances M, Rendueles M, Díaz M (2013) Alkaline hydrolysis of porcine blood haemoglobin: applications for peptide and amino acid production. Animal Production Science 53, 121–128.
Alkaline hydrolysis of porcine blood haemoglobin: applications for peptide and amino acid production.Crossref | GoogleScholarGoogle Scholar |

AOAC International (2000) ‘Official methods of analysis of the AOAC International.’ 17th edn. (AOAC International: Rockville, MD, USA)

ASABE (2008) ‘Standard ANSI/ASAE S319.4: method of determining and expressing fineness of feed materials by sieving.’ (American Society of Agricultural and Biological Engineers: St. Joseph, MI, USA)

Apple JK, Davis JC, Stephenson J (1999) Influence of body condition score on by-product yield and value from cull beef cows. Journal of Animal Science 77, 2670–2679.
Influence of body condition score on by-product yield and value from cull beef cows.Crossref | GoogleScholarGoogle Scholar | 10521026PubMed |

Barshan S, Khalaji S, Hedayati M, Yari M (2019) Influence of bone meal degelatinisation and calcium source and particle size on broiler performance, bone characteristics and digestive and plasma alkaline phosphatase activity. British Poultry Science 60, 297–308.
Influence of bone meal degelatinisation and calcium source and particle size on broiler performance, bone characteristics and digestive and plasma alkaline phosphatase activity.Crossref | GoogleScholarGoogle Scholar | 30836768PubMed |

Bautista J, Corpas R, Cremades O, Hernandez-Pinzon I, Romos R, Villaneuva A (2000) Sunflower protein hydrolysates for dietary treatment of patients with liver failure. Journal of the American Oil Chemists’ Society 77, 121–126.
Sunflower protein hydrolysates for dietary treatment of patients with liver failure.Crossref | GoogleScholarGoogle Scholar |

Bedford M, Classen H (1993) An in vitro assay for prediction of broiler intestinal viscosity and growth when fed rye-based diets in the presence of exogenous enzymes. Poultry Science 72, 137–143.
An in vitro assay for prediction of broiler intestinal viscosity and growth when fed rye-based diets in the presence of exogenous enzymes.Crossref | GoogleScholarGoogle Scholar | 8426842PubMed |

Beyranvand F, Khalaji S, Zamani A, Manafi M (2019) Effects of gelatin prepared from calf bones rich in phosphorus on broiler performance, bone characteristics and digestive enzymes activity. British Poultry Science 60, 31–38.
Effects of gelatin prepared from calf bones rich in phosphorus on broiler performance, bone characteristics and digestive enzymes activity.Crossref | GoogleScholarGoogle Scholar | 30309249PubMed |

Borda E, Martinez-Puig D, Perez JF (2005) An hydrolyzed protein concentrate (Palbio 62R) increases feed intake and villus height in early weaning pigs. In ‘Proceedings 56th annual meeting of the European Association for Animal Production’. Uppsala, Sweden. p. 154. (Abstr.) (European Federation of Animal Science: Rome)

Bryant MP, Burkey LA (1953) Cultural methods and some characteristics of some of the more numerous groups of bacteria in the bovine rumen. Journal of Dairy Science 36, 205–217.
Cultural methods and some characteristics of some of the more numerous groups of bacteria in the bovine rumen.Crossref | GoogleScholarGoogle Scholar |

Cahu CL, Zambonino Infante JL, Escaffre AM, Bergot P, Kaushik S (1998) Preliminary results on sea bass (Dicentrarchus labrax) larvae rearing with compound diet from first feeding. Comparison with carp (Cyprinus carpio) larvae. Aquaculture 169, 1–7.
Preliminary results on sea bass (Dicentrarchus labrax) larvae rearing with compound diet from first feeding. Comparison with carp (Cyprinus carpio) larvae.Crossref | GoogleScholarGoogle Scholar |

Carpenter JW, Palmer AZ, Kirk WG, Peacock FM, Koger M (1961) Slaughter and carcass characteristics of Brahman and Brahman–Shorthorn crossbred steers. Journal of Animal Science 20, 336–340.

Chatterjee C, Gleddie S, Xiao CW (2018) Soybean bioactive peptides and their functional properties. Nutrients 10, 1211
Soybean bioactive peptides and their functional properties.Crossref | GoogleScholarGoogle Scholar |

Clemente A (2000) Enzymatic protein hydrolysates in human nutrition. Trends in Food Science & Technology 11, 254–262.
Enzymatic protein hydrolysates in human nutrition.Crossref | GoogleScholarGoogle Scholar |

Corassa A, Lopes DC, Pena S, de Freitas LS, Pena GM (2007) Hydrolyzed of intestinal mucosa of swine in substitution to blood plasma in diets of pigs from 21 to 49 days. Revista Brasileira de Zootecnia 36, 2029–2036.
Hydrolyzed of intestinal mucosa of swine in substitution to blood plasma in diets of pigs from 21 to 49 days.Crossref | GoogleScholarGoogle Scholar |

De Angelis E, Pilolli R, Bavaro SL, Monaci L (2017) Insight into the gastro-duodenal digestion resistance of soybean proteins and potential implications for residual immunogenicity. Food & Function 8, 1599–1610.
Insight into the gastro-duodenal digestion resistance of soybean proteins and potential implications for residual immunogenicity.Crossref | GoogleScholarGoogle Scholar |

Erlanger BF, Kokawskyo N, Cohen W (1961) The preparation and properties of two new chromogenic substances of trypsin. Archives of Biochemistry and Biophysics 95, 271–278.
The preparation and properties of two new chromogenic substances of trypsin.Crossref | GoogleScholarGoogle Scholar | 13890599PubMed |

Fossati P, Pricipe L, Berti G (1980) Use of 3,5-dichloro-2-hydroxybenzene sulfonic acid/4-aminophenazone chromogenic system in the direct enzymic assay of uric acid in serum and urine. Clinical Chemistry 26, 227–231.
Use of 3,5-dichloro-2-hydroxybenzene sulfonic acid/4-aminophenazone chromogenic system in the direct enzymic assay of uric acid in serum and urine.Crossref | GoogleScholarGoogle Scholar | 7353268PubMed |

Fountoulakis M, Hans-Werner L (1998) Hydrolysis and amino acid composition analysis of proteins. Journal of Chromatography. A 826, 109–134.
Hydrolysis and amino acid composition analysis of proteins.Crossref | GoogleScholarGoogle Scholar | 9917165PubMed |

Frikha M, Mohiti-Asli M, Chetrit C, Mateos GG (2014) Hydrolyzed porcine mucosa in broiler diets: effects on growth performance, nutrient retention, and histomorphology of the small intestine. Poultry Science 93, 400–411.
Hydrolyzed porcine mucosa in broiler diets: effects on growth performance, nutrient retention, and histomorphology of the small intestine.Crossref | GoogleScholarGoogle Scholar | 24570462PubMed |

Gilbert ER, Wong EA, Webb KE (2008) Board-invited review: peptide absorption and utilization: implications for animal nutrition and health. Journal of Animal Science 86, 2135–2155.
Board-invited review: peptide absorption and utilization: implications for animal nutrition and health.Crossref | GoogleScholarGoogle Scholar | 18441086PubMed |

Gómez-Guillén MC, López-Caballero ME, López de Lacey A, Alem?n A, Giménez B, Montero P (2010) Antioxidant and antimicrobial peptide fractions from squid and tuna skin gelatin. In ‘Sea by-products as a real material: new ways of application’. Ch. 7, pp. 89–115. (Transworld Research Network Signpost: Thiruvananthapuram, India)

Hong KJ, Lee CH, Kim SW (2004) Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals. Journal of Medicinal Food 7, 430–435.
Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals.Crossref | GoogleScholarGoogle Scholar | 15671685PubMed |

Hou Y, Wu Z, Dai Z, Wang G, Wu G (2017) Protein hydrolysates in animal nutrition: industrial production, bioactive peptides, and functional significance. Journal of Animal Science and Biotechnology 8, 24
Protein hydrolysates in animal nutrition: industrial production, bioactive peptides, and functional significance.Crossref | GoogleScholarGoogle Scholar | 28286649PubMed |

Hoyle NT, Merritt JH (1994) Quality of fish protein hydrolysates from herring (Clupea harengus). Journal of Food Science 59, 76–79.
Quality of fish protein hydrolysates from herring (Clupea harengus).Crossref | GoogleScholarGoogle Scholar |

Iranian Council of Animal Care (1995) ‘Guide to the care and use of experimental animals.’ Vol. 1. (Isfahan University of Technology: Isfahan, Iran)

Johnston-Banks FA (1990) Gelatin. In ‘Food gels’. pp. 233–289. (Elsevier Applied Food Science: New York) Series.

Jus S, Kokol V, Guebitz GM (2009) Tyrosinase-catalysed coating of wool fibres with different protein-based biomaterials. Journal of Biomaterials Science, Polymer Edition 20, 253–269.
Tyrosinase-catalysed coating of wool fibres with different protein-based biomaterials.Crossref | GoogleScholarGoogle Scholar |

Khalaji S, Manafi M, Olfati Z, Hedayati M, Latifi M, Veysi A (2016) Replacing soybean meal with gelatin extracted from cow skin and corn protein concentrate as a protein source in broiler diets. Poultry Science 95, 287–297.
Replacing soybean meal with gelatin extracted from cow skin and corn protein concentrate as a protein source in broiler diets.Crossref | GoogleScholarGoogle Scholar | 26574036PubMed |

Khiari Z, Rico D, Martin-Diana AB, Barry-Ryan C (2013) Comparison between gelatines extracted from mackerel and blue whiting bones after different pre-treatments. Food Chemistry 139, 347–354.
Comparison between gelatines extracted from mackerel and blue whiting bones after different pre-treatments.Crossref | GoogleScholarGoogle Scholar | 23561116PubMed |

Kotzamanis YP, Gisbert E, Gatesoupe FJ, Zambonino Infante J, Cahu C (2007) Effects of different dietary levels of fish protein hydrolysates on growth, digestive enzymes, gut microbiota and resistance to Vibrio anguillarum in European sea bass (Dicentrarchus labrax) larvae. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 147, 205–214.
Effects of different dietary levels of fish protein hydrolysates on growth, digestive enzymes, gut microbiota and resistance to Vibrio anguillarum in European sea bass (Dicentrarchus labrax) larvae.Crossref | GoogleScholarGoogle Scholar |

Kunitz M (1947) Crystalline soybean trypsin inhibitor: II. General properties. The Journal of General Physiology 30, 291–310.
Crystalline soybean trypsin inhibitor: II. General properties.Crossref | GoogleScholarGoogle Scholar | 19873496PubMed |

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Crossref | GoogleScholarGoogle Scholar | 5432063PubMed |

Liu XQ, Yonekura M, Tsutsumi M, Sano Y (1996) Physicochemical properties of aggregates of globin hydrolysates. Journal of Agricultural and Food Chemistry 44, 2957–2961.
Physicochemical properties of aggregates of globin hydrolysates.Crossref | GoogleScholarGoogle Scholar |

Liu TX, Wang J, Zhao MM (2010) In vitro haem solubility of red cell fraction of porcine blood under various treatments. International Journal of Food Science & Technology 45, 719–725.
In vitro haem solubility of red cell fraction of porcine blood under various treatments.Crossref | GoogleScholarGoogle Scholar |

Lowry OH, Rosebiough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. The Journal of Biological Chemistry 193, 265–275.

Mathies CJ (1964) Adaptation of the berthelot color reaction for the determination of urea nitrogen in serum and urine to an ultramicro system. Clinical Chemistry 10, 366–369.
Adaptation of the berthelot color reaction for the determination of urea nitrogen in serum and urine to an ultramicro system.Crossref | GoogleScholarGoogle Scholar |

Ozols J (1990) ‘Methods in enzymology.’ Vol. 182. (Academic Press: San Diego, CA, USA)

Ravindran G, Bryden WL (2005) Tryptophan determination in proteins and feedstuffs by ion exchange chromatography. Food Chemistry 89, 309–314.
Tryptophan determination in proteins and feedstuffs by ion exchange chromatography.Crossref | GoogleScholarGoogle Scholar |

Rendueles M, Moure F, Fernandez A, Díaz M (1997) Preliminary studies on the processing of slaughter-house blood for protein recovery. Resource and Environmental Biotechnology 1, 193–206.

Sims JT, Bailey AJ (1992) Quantitative analysis of collagen and elastin crosslinks using a single-column system. Journal of Chromatography. A 582, 49–55.
Quantitative analysis of collagen and elastin crosslinks using a single-column system.Crossref | GoogleScholarGoogle Scholar |

Singh BP, Vij S, Hati S (2014) Functional significance of bioactive peptides derived from soybean. Peptides 54, 171–179.
Functional significance of bioactive peptides derived from soybean.Crossref | GoogleScholarGoogle Scholar | 24508378PubMed |

Udenigwe CC, Aluko RE (2012) Food protein-derived bioactive peptides: production, processing, and potential health benefits. Journal of Food Science 77, R11–R24.
Food protein-derived bioactive peptides: production, processing, and potential health benefits.Crossref | GoogleScholarGoogle Scholar | 22260122PubMed |

Ulfa M, Trisunaryanti W, Falah II, Kartini I (2015) Characterisation of gelatines extracted from cow bone for carbon synthesis. Journal of Applied Chemistry 8, 57–63.

Vaghefi N, Nedjaoum F, Guilochon D, Bureau F, Arhan P, Bouglé D (2002) Influence of the extent of haemoglobin hydrolysis on the digestive absorption of heme iron. An in vitro study. Journal of Agricultural and Food Chemistry 50, 4969–4973.
Influence of the extent of haemoglobin hydrolysis on the digestive absorption of heme iron. An in vitro study.Crossref | GoogleScholarGoogle Scholar | 12166991PubMed |

Vente-Spreeuwenberg MAM, Verdonk JMAJ, Koninkx JFJG, Beynen AC, Verstegen MWA (2004) Dietary protein hydrolysates vs. the intact proteins do not enhance mucosal integrity and growth performance in weaned piglets. Livestock Production Science 85, 151–164.
Dietary protein hydrolysates vs. the intact proteins do not enhance mucosal integrity and growth performance in weaned piglets.Crossref | GoogleScholarGoogle Scholar |

Vijayalakshmi IMA, Lemieux L, Amoit J (1986) High performance size exclusion liquid chromatography of small molecular weight peptides from protein hydrolysates using methanol as a mobile phase additive. Journal of Liquid Chromatography 9, 3559–3576.
High performance size exclusion liquid chromatography of small molecular weight peptides from protein hydrolysates using methanol as a mobile phase additive.Crossref | GoogleScholarGoogle Scholar |

Walter K, Schutt C (1974) Alkaline phosphatase in serum (continuous assay). In ‘Methods of enzymatic analysis’. 2nd edn. Vol. 2. (Ed. HU Bergmeyer) pp. 860–864. (Academic Press: New York)

Zambonino Infante JL, Cahu CL, Peres A (1997) Partial substitution of di- and tripeptides for native proteins in sea bass diet improves Dicentrarchus labrax larval development. The Journal of Nutrition 127, 608–614.
Partial substitution of di- and tripeptides for native proteins in sea bass diet improves Dicentrarchus labrax larval development.Crossref | GoogleScholarGoogle Scholar | 9109612PubMed |