ISSN 2308-4057 (Print),
ISSN 2310-9599 (Online)

Mechanically activated hydrolysis of plant-derived proteins in food industry

Abstract
A poor consumption of important nutrients triggered a public interest in functional foods that contain easy-to-digest proteins. The present research features fractionation, mechanical activation, and enzymatic hydrolysis of pea protein. According to modern chemical methods, the protein content in the original pea biomass was 24.3% and its molecular weight distribution (MWD) was 5–135 kDa. Fractionation, or protein displacement, resulted in four fractions of biopolymers with different chemical composition, i.e. a different content of protein and carbohydrate molecules. The paper introduces some data on the enzymatic transformations of the substrate. A set of experiments made it possible to define the optimal conditions for the mechanical activation of pea biomass with proteolytic enzymes. The enzymes were obtained from Protosubtilin G3x, a complex enzyme preparation. When the substrate and the enzymes were mechanically activated together, it produced mechanocomposite, an intermediate product with increased reactivity. It increased the specific surface area by 3.2 times and doubled the crystallinity of the substrate. As a result, the rate and yield of the subsequent enzymatic hydrolysis increased from 18% to 61%. The study determined the capacity of the substrate in relation to the enzyme preparation. Under optimal conditions, the pea hydrolysis destroyed protein molecules within two hours. After four hours of hydrolysis, no changes were detected. A polyacrylamide gel electrophoresis revealed non-hydrolysed protein molecules with MW ≈ 20 kDa. Presumably, they corresponded with legumin, which is resistant to neutral and alkaline proteases. The resulting hydrolysates were spray-dried to test their potential use as a food component. The product obtained by spray-drying had a monomodal distribution of particle sizes of spherical shape with adiameter of 5–20 μm.
Keywords
Mechanochemistry, mechanochemical activation, mechanocomposite, plant materials, enzymatic hydrolysis, destruction of protein molecules, polypeptides, amino acids, spray-drying
REFERENCES
  1. European Commission. Horizon 2020: Annual Monitoring Report. Publications Office of the European Union; 2015. DOI: https://doi.org/10.2777/32.
  2. Ulberth F, Maragkoudakis P, Czimbalmos A, Wollgast J, Rzychon M, Caldeira S, et al. Tomorrow’s healthy society. Research priorities for foods and diets: final report – Study. Luxemburg: Publications Office of the European Union; 2014. 116 p. DOI: https://doi.org/10.2788/1395.
  3. Bannikova AV, Evdokimov IA. The scientific and practical principles of creating products with increased protein content. Foods and Raw Materials. 2015;3(2):3–12. DOI: https://doi.org/10.12737/13114.
  4. Salvatore S, Vandenplas Y. Hydrolyzed Proteins in Allergy. In: Bhatia J, Shamir R, Vandenplas Y, editors. Protein in Neonatal and Infant Nutrition: Recent Updates. Basel: Nestec; 2016. pp. 11–27. DOI: https://doi.org/10.1159/000442699.
  5. Barać M, Cabrilo S, Pešić M, Stanojević S, Pavlićević M, Maćej O, et al. Functional properties of pea (Pisum sativum, L.) protein isolates modified with chymosin. International Journal of Molecular Sciences. 2011;12(12):8372–8387. DOI: https://doi.org/10.3390/ijms12128372.
  6. Ozuna C, León-Galván MF. Cucurbitaceae Seed Protein Hydrolysates as a Potential Source of Bioactive Peptides with Functional Properties. BioMed Research International. 2017;2017. DOI: https://doi.org/10.1155/2017/2121878.
  7. Sandberg A-S. Developing functional ingredients: a case study of pea protein. In: Saarela M, editor. Functional Foods: Concept to Product: Second Edition. Woodhead Publishing; 2011. pp. 358–382. DOI: https://doi.org/10.1533/9780857092557.3.358.
  8. Daniells S. US pea protein market ‘ready to explode’ [Internet]. [cited 2018 Sep 10]. Available from: https://www. foodnavigator-usa.com/Article/2013/01/29/US-pea-protein-market-ready-to-explode.
  9. Nuzest Clean Lean Protein - Premium Vegan Protein Powder, Plant Protein Powder, European Golden Pea Protein, Dairy Free, Gluten Free, GMO Free, Naturally Sweetened, Chai Turmeric Maca, 9 SRV, 7.9 oz [Internet]. [cited 2018 Sep 10]. Available from: https://www.amazon.com/Nuzest-Clean-Lean-Protein-Functional/dp/B0776NJ7QC?th=1.
  10. Barac MB, Pešić MB, Stanojevic SP, Kostić AZ, Bivolarevic V. Comparative study of the functional properties of three legume seed isolates: adzuki, pea and soy bean. Journal of Food Science and Technology. 2015;52(5):2779–2787. DOI: https://doi.org/10.1007/s13197-014-1298-6.
  11. Mathai J, Liu Y, Stein H. Values for digestible indispensable amino acid scores (DIAAS) for some dairy and plant proteins may better describe protein quality than values calculated using the concept for protein digestibility-corrected amino acid scores (PDCAAS). British Journal of Nutrition. 2017;117(4):490–499. DOI: https://doi.org/10.1017/S0007114517000125.
  12. Jung S, Lamsal BP, Stepien V, Johnson LA, Murphy PA. Functionality of soy protein produced by enzyme-assisted extraction. Journal of the American Oil Chemists’ Society. 2006;83(1):71–78. DOI: https://doi.org/10.1007/s11746-006-1178-y.
  13. Gueguen J, Cerletti P. Protein of some legume seeds: soybean, pea, fababean and lupin. In: Hudson BJF, editor. New and Developing Sources of Food Proteins. Boston: Springer; 1994. pp. 145–193. DOI: https://doi.org/10.1007/978-1-4615-2652-0_6.
  14. Kriger OV, Kashirskih EV, Babich , Noskova SYu. Oat Protein Concentrate Production. Foods and Raw Materials. 2018;6(1):47–55. DOI: https://doi.org/10.21603/2308-4057-2018-1-47-55.
  15. Lomovsky O, Bychkov A, Lomovsky I. Mechanical Pretreatment. In: Mussatto SI, editor. Biomass Fractionation Technologies for a Lignocellulosic Feedstock Based Biorefinery. Elsevier; 2016. pp. 23–55. DOI: https://doi.org/10.1016/B978-0-12-802323-5.00002-5.
  16. Kamarludin S, Jainal S, Azizan M, Safaai AM, Sharliza N, Daud M, et al. Mechanical Pretreatment of Lignocellulosic Biomass for Biofuel Production. Applied Mechanics and Materials. 2014;625:838–841. DOI: https://doi.org/10.4028/www.scientific.net/AMM.625.838.
  17. Podgorbunskikh EM, Bychkov AL, Bulina NV, Lomovskii OI. Disordering of the Crystal Structure of Cellulose Under Mechanical Activation. Journal of Structural Chemistry. 2018;59(1):201–208. DOI: https://doi.org/10.1134/S0022476618010328.
  18. State Standard 23636-90. Fermentative preparation protosubtilin G3x. Specifications. Moscow: Standards Publishing; 1990. 7 p.
  19. ISO 6496. Animal feeding stuffs – Determination of moisture and other volatile matter content. Geneve: International Organization for Standardization; 1999. 6 .
  20. ISO 5984. Animal feeding stuffs -- Determination of crude ash. Geneve: International Organization for Standardization; 2002. 6 .
  21. Gregg S, Sing K. Adsorption, surface area and porosity. London: Academic Press; 1967. 408 p.
  22. Fadeeva VP, Tikhova VD, Nikulicheva ON. Elemental analysis of organic compounds with the use of automated CHNS analyzers. Journal of Analytical Chemistry. 2008;63(11):1094–1106. DOI: https://doi.org/10.1134/S1061934808110142.
  23. Sáez-Plaza P, Michałowski T, Navas MJ, Asuero AG, Wybraniec S. An Overview of the Kjeldahl Method of Nitrogen Determination. Part I. Early History, Chemistry of the Procedure, and Titrimetric Finish. Critical Reviews in Analytical Chemistry. 2013;43(4):178–223. DOI: https://doi.org/10.1080/10408347.2012.751786.
  24. Salo-Väänänen PP, Koivistoinen PE. Determination of protein in foods: Comparison of net protein and crude protein (N x 6.25) values. Food Chemistry. 1996;57(1):27–31. DOI: https://doi.org/10.1016/0308-8146(96)00157-4.
  25. Chace DH, Kalas T, Naylor EW. Use of Tandem Mass Spectrometry for Multianalyte Screening of Dried Blood Specimens from Newborns. Clinical Chemistry. 2003;49(11):1797–1817. DOI: https://doi.org/10.1373/clinchem.2003.022178.
  26. Gallagher S.R. One-Dimensional SDS Gel Electrophoresis of Proteins. Current Protocols in Protein Science. 2012;68(1). DOI: https://doi.org/10.1002/0471140864.ps1001s68.
  27. Dyballa N, Metzger S. Fast and sensitive colloidal coomassie G-250 staining for proteins in polyacrylamide gels. Journal of Visualized Experiments. 2009;30. DOI: https://doi.org/10.3791/1431.
  28. Mikhailova K, Krasikov V, Malahova I, Kozmin Y, Kalambet Y. Data processing in planar chromatography and gel electrophoresis using ‘Chrom&Spec’ software. Analytics. 2014;19(6):56–61.
  29. Talab HA. Starch-Protein Extraction and Separation it from Green Pea. European Online Journal of Natural and Social Sciences. 2016;5(4):1012–1017.
  30. Swanson BG. Pea and lentil protein extraction and functionality. Journal of the American Oil Chemists’ Society. 1990;67(5):276–280. DOI: https://doi.org/10.1007/BF02539676.
  31. Byulleteni o sostoyanii selʹskogo khozyaystva (ehlektronnye versii) [Bulletins on the state of agriculture (e-versions)] [Internet]. [cited 2018 Sep 10]. Available from: www.gks.ru/wps/wcm/connect/rosstat_main/rosstat/ru/statistics/publications/catalog/doc_1265196018516.
  32. Skurikhin IM, Volgarev MN. Khimicheskiy sostav pishchevykh produktov. Kn. 2: Spravochnye tablitsy soderzhaniya aminokislot, zhirnykh kislot, vitaminov, makro - i mikro - ehlementov, organicheskikh kislot i uglevodov [Chemical composition of food. Book 2: Reference tables of the contents of amino acids, fatty acids, vitamins, macro- and microelements, organic acids, and carbohydrates]. Moscow: Agropromizdat; 1987. 360 p. (In Russ.).
  33. Mession J-L, Sok N, Assifaoui A, Saurel R. Thermal Denaturation of Pea Globulins (Pisum sativum L.) - Molecular Interactions Leading to Heat-Induced Protein Aggregation. Journal of Agricultural and Food Chemistry. 2013;61(6):1196–1204. DOI: https://doi.org/10.1021/jf303739n.
  34. Wu YV, Nicho1s NN. Fine Grinding and Air Classification of Field Pea. Cereal Chemistry. 2005;82(3):341–344. DOI: https://doi.org/10.1094/CC-82-0341.
  35. Bychkov AL, Lomovsky OI. Recent advances in mechanochemical processing of plant raw materials. Chemistry of plant raw material. 2017;(2):35–47. (In Russ.). DOI: https://doi.org/10.14258/jcprm.2017021546.
  36. Kuznetsov BN, Chesnokov NV, Yatsenkova OV, Sharypova VI. New methods of heterogeneous catalysis for lignocellulosic biomass conversion to chemicals. Russian Chemical Bulletin. 2013;62(7):1493–1502. DOI: https://doi.org/10.1007/s11172-013-0213-z.
  37. Chen C, Chi Y-J, Xu W. Comparisons on the Functional Properties and Antioxidant Activity of Spray-Dried and Freeze-Dried Egg White Protein Hydrolysate. Food and Bioprocess Technology. 2012;5(6):2342–2352. DOI: https://doi.org/10.1007/s11947-011-0606-7.
  38. Zhao Q, Xiong H, Selomulya C, Chen X, Huang S, Ruan X, et al. Effects of Spray Drying and Freeze Drying on the Properties of Protein Isolate from Rice Dreg Protein. Food and Bioprocess Technology. 2012;6(7):1759–1769. DOI: https://doi.org/10.1007/s11947-012-0844-3.
  39. Pelgrom PJM, Vissers AM, Boom RM, Schutyser MAI. Dry fractionation for production of functional pea protein concentrates. Food Research International. 2013;53(1):232–239. DOI: https://doi.org/10.1016/j.foodres.2013.05.004.
  40. Archaina D, Leiva G, Salvatori D, Schebor C. Physical and functional properties of spray-dried powders from blackcurrant juice and extracts obtained from the waste of juice processing. Food Science and Technology International. 2017;24(1):78–86. DOI: https://doi.org/10.1177/1082013217729601.
How to quote?
Gavrilova KV, Bychkov AL, Bychkova ES, Akimenko ZA, Chernonosov AA, Kalambet YuA, et al. Mechanically activated hydrolysis of plant-derived proteins in food industry. Foods and Raw Materials. 2019;7(2):255–263. DOI: http://doi.org/10.21603/2308-4057-2019-2-255-263
About journal

Download
Contents
Abstract
Keywords
References