Protein Fractionation of Green Leaves as an Underutilized Food Source—Protein Yield and the Effect of Process Parameters

Foods - Tập 10 Số 11 - Trang 2533
Anna-Lovisa Nynäs1, William R. Newson1, Eva Johansson1
1Department of Plant Breeding, Swedish University of Agricultural Sciences, P.O. Box 190, SE-23422 Lomma, Sweden

Tóm tắt

Green biomass has potential as a sustainable protein source for human consumption, due to its abundance and favorable properties of its main protein, RuBisCO. Here, protein fractionation outcomes of green leafy biomass from nine crops were evaluated using a standard protocol with three major steps: juicing, thermal precipitation, and acid precipitation. Successful protein fractionation, with a freeze-dried, resolubilized white protein isolate containing RuBisCO as the final fraction, was achieved for seven of the crops, although the amount and quality of the resulting fractions differed considerably between crops. Biomass structure was negatively correlated with successful fractionation of proteins from biomass to green juice. The proteins in carrot and cabbage leaves were strongly associated with particles in the green juice, resulting in unsuccessful fractionation. Differences in thermal stability were correlated with relatedness of the biomass types, e.g., Beta vulgaris varieties showed similar performance in thermal precipitation. The optimal pH values identified for acid precipitation of soluble leaf proteins were lower than the theoretical value for RuBisCO for all biomass types, but with clear differences between biomass types. These findings reveal the challenges in using one standard fractionation protocol for production of food proteins from all types of green biomass and indicate that a general fractionation procedure where parameters are easily adjusted based on biomass type should instead be developed.

Từ khóa


Tài liệu tham khảo

Barclays (2020, July 06). Carving Up the Alternative Meat Market. Available online: https://www.investmentbank.barclays.com/our-insights/carving-up-the-alternative-meat-market.html.

Smart Protein Project (2021, October 12). Plant-Based Foods in Europe: How Big Is the Market? Smart Protein Plant-Based Food Sector Report by Smart Protein Project, European Union’s Horizon 2020 Research and Innovation Programme (No 862957). Available online: https://knowledge4policy.ec.europa.eu/publication/plant-based-foods-europe-how-big-market_en.

Furu, M. (2020, March 27). Marknadsanalys och Potential för Växtbaserade Proteiner. Available online: https://www.lrf.se/globalassets/dokument/mitt-lrf/nyheter/2020/marknadsanalys-och-potential-for-vaxtbaserade-proteiner.pdf.

USDA (2021). Oilseeds: World Markets and Trade, United States Department of Agriculture: Economics, Statistics and Market Information System. Available online: https://usda.library.cornell.edu/concern/publications/tx31qh68h?locale=en.

Sozer, 2015, Plant protein ingredients for future foods, Agro FOOD Ind. Hi-Tech, 26, 56

Pirie, 1942, The direct use of leaf protein in human nutrition, Chem. Ind., 61, 45

Agyei, 2018, Plant RuBisCo: An underutilized protein for food applications, J. Am. Oil Chem. Soc., 95, 1063, 10.1002/aocs.12104

Alting, 2011, From waste product to food ingredient: The extraction of abundant plant protein RuBisCo, New Food, 14, 10

Berndtsson, E., Nynäs, A.-L., Newson, W.R., Langton, M., Andersson, R., Johansson, E., and Olsson, M.E. (2019). The underutilised side streams of broccoli and kale–valorisation via proteins and phenols. Sustainable Governance and Management of Food Systems: Ethical Perspectives, Wageningen Academic Publishers.

Andersson, 2008, Structure and function of Rubisco, Plant Physiol. Biochem., 46, 275, 10.1016/j.plaphy.2008.01.001

Martin, 2019, Comparison of the functional properties of RuBisCO protein isolate extracted from sugar beet leaves with commercial whey protein and soy protein isolates, J. Sci. Food Agric., 99, 1568, 10.1002/jsfa.9335

Lamsal, 2007, Some physicochemical and functional properties of alfalfa soluble leaf proteins, LWT—Food Sci. Technol., 40, 1520, 10.1016/j.lwt.2006.11.010

Selling, 2016, Extraction, composition, and functional properties of dried alfalfa (Medicago sativa L.) leaf protein, J. Sci. Food Agric., 97, 882

Patel, 2008, Rubisco gene expression in C4 plants, J. Exp. Bot., 59, 1625, 10.1093/jxb/erm368

Wang, 1975, Composition of alfalfa leaf protein isolates, J. Food Sci., 40, 1156, 10.1111/j.1365-2621.1975.tb01041.x

Merodio, 1987, Preparation and properties of a white protein fraction in high yield from sugar beet (Beta vulgaris L.) leaves, J. Sci. Food Agric., 44, 237, 10.1002/jsfa.2740440305

Knuckles, 1982, Functional properties of edible protein concentrates from alfalfa, J. Agric. Food Chem., 30, 748, 10.1021/jf00112a030

Gieteling, 2016, Recovery of protein from green leaves: Overview of crucial steps for utilisation, Food Chem., 203, 402, 10.1016/j.foodchem.2016.02.092

Edwards, 1975, Pilot plant production of an edible white fraction leaf protein concentrate from alfalfa, J. Agric. Food Chem., 23, 620, 10.1021/jf60200a046

Martin, 2014, Characterization of heat-set gels from RuBisCO in comparison to those from other proteins, J. Agric. Food Chem., 62, 10783, 10.1021/jf502905g

Sheen, 1991, Comparison of chemical and functional properties of soluble leaf proteins from four plant species, J. Agric. Food Chem., 39, 681, 10.1021/jf00004a011

Amer, B., Juul, L., Møller, A., Møller, H., and Dalsgaard, T. (2020). Improved solubility of proteins from white and red clover–Inhibition of redox enzymes. Int. J. Food Sci. Technol., in press.

Bray, 1979, Preparation of white leaf protein concentrate using a polyanionic flocculant, J. Sci. Food Agric., 30, 171, 10.1002/jsfa.2740300212

Fiorentini, 1981, Pilot plant production of an edible alfalfa protein concentrate, J. Food Sci., 46, 1514, 10.1111/j.1365-2621.1981.tb04209.x

Colas, 2013, Green crop fractionation by twin-screw extrusion: Influence of the screw profile on alfalfa (Medicago sativa) dehydration and protein extraction, Chem. Eng. Process. Process Intensif., 72, 1, 10.1016/j.cep.2013.05.017

Miller, 1973, Centrifugal separation of white and green protein fractions from alfalfa juice following controlled heating, J. Agric. Food Chem., 21, 886, 10.1021/jf60189a020

Martinez, 1998, Production and chemical composition of alfalfa protein concentrate obtained by freezing, Anim. Feed Sci. Technol., 72, 169, 10.1016/S0377-8401(97)00167-3

Jwanny, 1993, Protein production for human use from Sugarbeet: Byproducts, Bioresour. Technol., 43, 67, 10.1016/0960-8524(93)90085-P

Kiskini, 2016, Effect of plant age on the quantity and quality of proteins extracted from sugar beet (Beta vulgaris L.) leaves, J. Agric. Food Chem., 64, 8305, 10.1021/acs.jafc.6b03095

Lamsal, 2003, Separation of protein fractions in alfalfa juice: Effects of some pre-treatment methods, Trans. ASAE, 46, 715, 10.13031/2013.13572

Merodio, 1983, Improved separation of green and soluble leaf proteins by pH shift, J. Agric. Food Chem., 31, 957, 10.1021/jf00119a009

Zhang, 2015, Leaf protein concentration of alfalfa juice by membrane technology, J. Membr. Sci., 489, 183, 10.1016/j.memsci.2015.03.092

Sedlar, 2021, Vegetable by-products as new sources of functional proteins, Plant Foods Hum. Nutr., 76, 31, 10.1007/s11130-020-00870-8

Walker, J.M. (2005). Protein identification and analysis tools on the ExPASy server. The Proteomics Protocols Handbook, Humana Press.

RStudio Team (2020). RStudio: Integrated Development for R. RStudio, RStudio, PBC.

Kuznetsova, 2017, lmerTest package: Tests in linear mixed effects models, J. Stat. Softw., 82, 1, 10.18637/jss.v082.i13

Miller, 1975, Soluble protein concentrate from alfalfa by low-temperature acid precipitation, J. Agric. Food Chem., 23, 1177, 10.1021/jf60202a037

Ytting, 2019, Influence of the development stage of perennial forage crops for the recovery yields of extractable proteins using lactic acid fermentation, J. Clean. Prod., 218, 1055, 10.1016/j.jclepro.2019.01.292

Bizot, 1993, Differential scanning calorimetric studies of the effects of ions and pH on ribulose 1, 5-bisphosphate carboxylase/oxygenase, Int. J. Biol. Macromol., 15, 195, 10.1016/0141-8130(93)90037-M

Betschart, 1973, Extractability and solubility of leaf protein, J. Agric. Food Chem., 21, 60, 10.1021/jf60185a019

Bhatia, S., Sharma, K., Dahiya, R., and Bera, T. (2015). Concepts and Techniques of Plant Tissue Culture Science. Modern Applications of Plant Biotechnology in Pharmaceutical Sciences, Academic Press. Chapter 4.

Kobbi, 2017, Purification and Recovery of RuBisCO Protein from Alfalfa Green Juice: Antioxidative Properties of Generated Protein Hydrolysate, Waste Biomass Valorization, 8, 493, 10.1007/s12649-016-9589-y

Firdaous, 2017, Adsorptive removal of polyphenols from an alfalfa white proteins concentrate: Adsorbent screening, adsorption kinetics and equilibrium study, Sep. Purif. Technol., 178, 29, 10.1016/j.seppur.2017.01.009

Tenorio, 2017, Processing concepts for the use of green leaves as raw materials for the food industry, J. Clean. Prod., 164, 736, 10.1016/j.jclepro.2017.06.248

Muneer, F., Hovmalm, H.P., Svensson, S.-E., Newson, W.R., Johansson, E., and Prade, T. (2021). Economic viability of protein concentrate production from green biomass of intermediate crops: A pre-feasibility study. J. Clean. Prod., 294.

Prade, T., Muneer, F., Berndtsson, E., Nynäs, A.-L., Svensson, S.-E., Newson, W.R., and Johansson, E. (2021). Protein fractionation of broccoli (Brassica oleracea, var Italica) and kale (Brassica oleracea, var. Sabellica) leaves—A pre-feasibility assessment and evaluation of fraction phenol and fibre content. Food Bioprod. Process., in press.