foods Article Development of High-Moisture Meat Analogues with Hemp and Soy Protein Using Extrusion Cooking Izalin Zahari 1,* , Ferawati Ferawati 2, Amanda Helstad 1, Cecilia Ahlström 1, Karolina Östbring 1 , Marilyn Rayner 1 and Jeanette K. Purhagen 1 1 Department of Food Technology Engineering and Nutrition, Lund University, Naturvetarvägen 12, 223 62 Lund, Sweden; [email protected] (A.H.); [email protected] (C.A.); [email protected] (K.Ö.); [email protected] (M.R.); [email protected] (J.K.P.) 2 Department of Chemistry and Biomedical Sciences, Linnaeus University, 392 31 Kalmar, Sweden; [email protected] * Correspondence: [email protected] Received: 30 April 2020; Accepted: 5 June 2020; Published: 11 June 2020 Abstract: The interest in plant-based products is growing in Western countries, mostly due to health and environmental issues that arise from the consumption and production of animal-based food products. Many vegan products today are made from soy, but drawbacks include the challenges of cultivating soy in colder climates such as northern Europe. Therefore, the present study investigates whether industrial hemp (Cannabis sativa) could substitute soy in the production of high moisture meat analogues (HMMA). A twin screw co-rotating extruder was used to investigate to what extent hemp protein concentrate (HPC) could replace soy protein isolate (SPI) in HMMAs. The substitution levels of HPC were 20 wt%, 40 wt% and 60 wt%. Pasting properties and melting temperature of the protein powders were characterized by Rapid Visco Analyzer (RVA) and Differential Scanning Calorimeter (DSC), respectively and the produced HMMA was analysed by determining the texture and colour attributes. The results showed that it is possible to extrude a mixture with up to 60% HPC. HPC absorbed less water and needed a higher denaturing temperature compared to SPI. Increasing the moisture content by 5% would have resulted in a reduction of hardness and chewiness. The lightness (L* value) was found to be significantly higher in SPI product and decreased in the mixture with higher HPC (p < 0.05). Keywords: high-moisture meat analogue; extrusion; hemp seed concentrate; soy protein isolate 1. Introduction The Western diet includes a high proportion of products from animal origins, and the high consumption of meat is a subject of various debates on its impact on human health and the environment [1]. High consumption of meat is linked to an increased risk of cardiovascular disease, type 2 diabetes, and different types of cancer [1]. Moreover, livestock production is responsible for 14.5% of global greenhouse gas emissions [2]. A shift towards a more plant-based diet is suggested to be one possible solution to reduce climate impact and create a healthier lifestyle [3,4]. As a consequence, there is a tremendous market growth in the plant-based food sector, driven by a growing number of consumers adopting a vegan, vegetarian, or flexitarian lifestyle [5]. Plant-based alternative products range from dairy analogues to meat substitutes. The meat substitutes market is still relatively small, but is projected to increase in the future. Europe leads the meat substitutes market globally, with 40% of the total market [5], which is predicted to reach ¿2.4 billion in 2025 as compared to ¿1.5 billion in 2018 [5]. The market is also expected to increase in Foods 2020, 9, 772; doi:10.3390/foods9060772 www.mdpi.com/journal/foods Foods 2020, 9, 772 2 of 13 Foods 2020, 9, x FOR PEER REVIEW 2 of 14 vegetableother regions, protein such are typical as North meat America substitutes and Asia on the [5]. market. Tofu, tempeh, Another seitan, type mycoprotein,of meat substitute, and texturized high- moisturevegetable meat protein analogue are (HMMA), typical meat is not substitutes widely av onailable the market.on the market Another yet; typethus, ofHMMA meat substitute,will be thehigh-moisture focus of the present meat analoguestudy. HMMA (HMMA), has a is desira not widelyble chewy available meat-like on the texture, market which yet; thus, together HMMA with will appropriatebe the focus flavouring of the present ingredients study. ca HMMAn offset has the a consumption desirable chewy of meat. meat-like texture, which together with appropriateThe vast majority flavouring of meat ingredients analogue can products offset the on consumption the European of market meat. are made from imported soybean,The while vast only majority a small of meat proportion analogue is productsmade from on other the European plant protein market sources are made such from as pea imported or mushroom.soybean, This while import only aof small raw material proportion could is madepotentially from reduce other plant the sustai proteinnable sources benefits such of asplant- pea or basedmushroom. products. This Therefore, import ofother raw crop materials that could could potentially be cultivated reduce over the a range sustainable of latitudes, benefits such of plant-basedas seeds fromproducts. industrial Therefore, hemp (Cannabis other crops sativa) that should could be beinvestigated. cultivated Optimization over a range of the latitudes, extrusion such process as seeds in orderfrom industrialto improve hemp the texture (Cannabis of sativaHMMA) should is one be of investigated. the most challenging Optimization aspects of thewhen extrusion it comes process to extrusionin order cooking. to improve There the have texture been of many HMMA estab islished one of extrusion the most challengingprocesses and aspects patents when for itHMMA comes to productionextrusion using cooking. soybean There [6–11], have but been only many a few established studies have extrusion been carried processes out on and formulations patents for based HMMA on productionplant proteins using other soybean than soy [6– 11[12–14].], but only a few studies have been carried out on formulations based onThe plant hemp proteins plant other has multifaceted than soy [12 –benefits14]. in many production sectors [15]. In Europe, hemp is mainly usedThe hempas a source plant of has fibre multifaceted for biomass, benefits textile, in and many the production rope industry sectors [15]. [However,15]. In Europe, there hempis an is increasingmainly usedinterest as ain source using of locally fibre for cultivated biomass, hemp textile, seeds and theas a rope raw industry material [15 for]. However,food production. there is an Hempincreasing seeds, the interest by-product in using from locally the cultivated fibre use for hemp the seedsconstruction as a raw industry, material forare foodrich in production. fats (25–35%, Hemp mostlyseeds, unsaturated the by-product fatty from acids), the proteins fibre use (20–25%), for the construction dietary fibre industry, (10–15%), are richvitamins, in fats and (25–35%, minerals mostly [16].unsaturated fatty acids), proteins (20–25%), dietary fibre (10–15%), vitamins, and minerals [16]. ProteinProtein is isthe the most most importantimportant ingredientingredient in thein the production production of HMMA of HMMA using extrusionusing extrusion technology. technology.During extrusion, During extrusion, protein is protein denatured, is denatured, unfolded, unfolded, realigned realigned and cross-linked, and cross-linked, due to the due shear, to the heat, shear,pressure heat, andpressure cooling and (Figure cooling1) achieved(Figure 1) in achiev the extrudered in the and extruder cooling and die, cooling respectively die, respectively [ 17]. The final [17].texture The final of HMMA texture is influencedof HMMA byis diinfluencedfferent types by ofdifferent protein bonds.types of Several protei factorsn bonds. determine Several thefactors protein determineinteractions the protein such as interactions protein type, such pre-treatment as protein type, and extrusionpre-treatment parameters and extrusion [17]. parameters [17]. FigureFigure 1. Changes 1. Changes in protein in protein conformation conformation during during extr extrusionusion of HMMA. of HMMA. Modified Modified illustration illustration adopted adopted fromfrom Zhang Zhang et al. et [17]. al. [17 ]. Previous works have shown that hemp seed protein in particular, has a high protein quality Previous works have shown that hemp seed protein in particular, has a high protein quality and and functionality [16,18,19]. However, no study uses hemp seed protein as a raw material for meat functionality [16,18,19]. However, no study uses hemp seed protein as a raw material for meat analogue production. Thus, this study was aimed to investigate the maximum ratio substitution of analogue production. Thus, this study was aimed to investigate the maximum ratio substitution of soy protein by hemp protein in the formulation of HMMA, as well as to study the characteristics of soy protein by hemp protein in the formulation of HMMA, as well as to study the characteristics of the HMMAs produced. the HMMAs produced. 2. Materials and Methods 2. Materials and Methods Soy protein isolate, SPI (91% protein, 1% carbohydrate, 3% fat, wet basis) and hemp protein Soy protein isolate, SPI (91% protein, 1% carbohydrate, 3% fat, wet basis) and hemp protein concentrate, HPC (71% protein, 15% carbohydrate, 7% fat, wet basis), were obtained from Bulk Powders concentrate, HPC (71% protein, 15% carbohydrate, 7% fat, wet basis), were obtained from Bulk Company and Natures Organic Food Company, Sweden, respectively (Figure2). Mixtures of hemp Powders Company and Natures Organic Food Company, Sweden, respectively (Figure 2). Mixtures