MINISTRY OF EDUCATION AND TRAINING HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY AND EDUCATION FACULTY FOR HIGH QUALITY TRAINING ACQUISITION OF PROTEIN FROM LIMA BEAN PHASEOLUS LUNATUS WITH
INTRODUCTION
Problem statement
The currently trend in human dietary habit involves the combination or partial replacement of plant-derived protein with animal-derived protein (Alves & Tavares 2019) This approach is gradually becoming popularity because there are some evidences can prove that a balanced diet incorporating both protein sources can contribute to lowering the risk of common human ailments, including cardiovascular disease, type 2 diabetes, and issues related to overweight and obesity (Ferrari et al., 2022)
Throughout history and up to the present day, the nutritional contributions from plants have consistently held a significant role in the daily diets of human beings Around the world, legumes play an important part in humanity diet Legume is considered the second most important food source for humans in a variety of locations of the world, after cereals, which are an essential part of the human diet Especially in developing countries , they serve as an affordable source of protein when animal protein is lacking Similar to grains and oil seeds, legumes are a great source of protein (22.37-28.05%) Fiber, vitamins, minerals and complex carbohydrates can all be found plentiful in legumes, which play an important role in human diet, contribute to the health of body (Du et al., 2014) Moreover, the food industry makes extensive use of some species of legumes, such soybean and common beans, because of their numerous functional qualities (like viscosity, emulsion and gelling abilities or foaming), which provide texture and sensory qualities (David Betancur-Ancona et al., 2004) Legumes also have a high lysine level, which makes them ideal for intergrating with other foods The nutrient-dense seeds have an important role in biodiversity, sustainable agriculture, food security, and minimizing environmental change (da Silva et al., 2009)
Lima bean (Phaseolus lunatus L) are a noteworthy member of the Legume family,
Fabaceae, a high protein content species range from 21-26% (M Sandoval-Peraza et al.,
2020), also known as “butter bean” in some regions, are renowned for their nutritional value and versatility in culinary applications The Lima bean seeds grow from specific areas will have distinct characteristics from others Originating from South America, particularly the Andes region, Lima beans have become a globally recognized legume
Nowadays, there are two popular types of protein: Protein Concentrate (PC) and Protein Isolate (PI) are available under powder form The way of deriving them are from alkaline-
2 based protein extraction processes PI is typically obtained through extraction in an alkaline environment followed by precipitation at the “isoelectric point”, while PC is acquired by extracting protein in an alkaline environment and subsequently precipitating it with
“alcohol” (da Silva et al., 2009; Zhao et al., 2012)
Moreover, plant-based protein sources offer valuable supplementation options for individuals whose following vegetarian or vegan diets Simultaneously, they adapt to the essential protein requirements of gymers or fitness enthusiasts who often depend on whey
For these reasons, there have been many studies around the world in testing different methods of recovering protein from plant sources, to meet the increasing needs of humans in using and consumiung recovery protein as a supplemental protein source Currently in Vietnam, protein products on the market are mainly produced from soybeans because of the popular crop, however Lima beans are not widely known by the populations and have high commercial consumption, even though they are also a nutritious source of protein from plants
Acknowledging that problem, our research teams have examined and experimented with many techniques across the worldwide on the legume family in order to implement and identify an appropriate procedure for Lima bean production, made with a high protein production recovery efficiency from Lima bean.
Research objective
The research objective of the thesis is to research, combine and totally complete the technological process, obtain protein products with high recovery efficiency Besides that, evaluate the physicochemical properties, functional properties of proteins from Lima bean seeds From there, propose product application ranges of Lima bean proteins in the food technology field.
Topic contents
The topic of “Acquisition of protein from Lima bean (Phaseolus lunatus) with high recovery efficiency” includes the following contents:
Synthesize scientific documents and articles, combine and experiment to select an appropriate technological process in producing protein from Lima bean with high collectiom efficiency
Conduct surveys and evaluate some product properties such as chemical compositions, structure and molecular weight of proteins obtained from Lima bean
Proposing directions for applying protein from Lima bean in the field of food technology.
Research object and Scope of research
Lima bean (Phaseolus lunatus) are provided by the Thu Duc Wholesale Agricultural Market (141 National Highway 1A, Tam Binh Ward, Thu Duc City, Vietnam)
Protein from Lima bean (Phaseolus lunatus)
The research was conducted at the laboratory of the Food Technology, Department of Chemical and Food Technology, Ho Chi Minh City University of Technical Education (HCMUTE).
Scientific and practical significance
The outcomes of the research contribute to enhancing the process of obtaining protein and offer technological parameters for conducting experimental researches on Lima bean protein production adapted to local conditions
The procedure for extracting protein from Lima bean can serve as a reference for research and academic studies at universities
The processing methodology for this product can be implemented either in pilot- scale or industrial-scale production, contributing to the expansion of protein product varieties available in the market
The results of the physicochemical and functional characteristics of protein help to determine the suitability of this product for large-scale commercial applications within food processing
LITERATURE REVIEW
Overview of material
One of the biggest families of flowering plants is the Fabaceae family, which includes legumes A double row of ovules on the typical legume pod serves as the family's uniting feature Flowers of this family are further distinguished by having five fused sepals and five petals The majority of the over 20,000 species capacity to adapt, especially to nutritionally deficient conditions, is aided by their ability to fix nitrogen through symbiotic relationships with bacteria belonging to the Rhizobiaceae family The current hypothesis postulates that a copy of the fundamental plant–mycorrhizal fungus identification system was utilised (or, to use evolutionary vernacular, "neofunc-tionalized") early in the evolution of legumes to identify nitrogen-fixing bacteria Above earth, a further process of co-evolution took place when bees and flowers adapted to one another for pollination Based on harvested area and global production, the legume family is the second most important in agriculture, behind the cereals (Poaceae), with over 650 million tons of grain legumes produced on 240 million ha in 2011 (MOD 2005) Legumes are credited with several roles that are frequently disregarded One-third of the plant protein and a comparable amount of the vegetable oil that is consumed by humans are found in grain legumes (Graham & Vance 2003) Legumes' complementary amino acid content to that of cereals and root crops may help to explain why the two families were domesticated together (Wang et al., 2003) Moreover, legumes are significant feed crops in tropical and temperate cLimates
Minerals that are necessary for consumption are provided by legumes (Grusak 2002) Furthermore, it has been suggested that the secondary metabolites that shield plants from pests and diseases may also shield humans from various types of cancer (Madar & Stark 2002) and may even help treat diabetes Grain legume eating has a hypoglycemic impact in addition to lowering blood cholesterol Trypsin inhibitors and allergens are examples of antinutritional elements that are secondary molecules (Skolnick et al., 2001)
The family has historically been split into three subfamilies: Papilionoideae, which includes the grain legumes in addition to the significant forage legumes, and Caesalpinioideae, which has 28 tribes The warm-season legumes (tribe Phaseoleae) are similarly closely linked (Lee et al., 2001), as are the cool-season legumes (tribes Fabeae,
Cicereae, and Trifolieae), which are somewhat separated from the Genisteae and closely related and extremely similar at the genome level (Wojciechowski et al., 2004) The Fabaceae family of plants, along with several other lesser-known species, are essential to biological nitrogen fixation (BNF), which occurs in symbiosis and is tens of times more efficient than in free-living bacteria The production of synthetic fertiliser began to fix more nitrogen than BNF only in the last quarter of the 20th century As Columella in ancient Rome demonstrated, even though ancient farmers would not have known how legumes worked, they would have recognised the influence of the bean on the next harvest (Evans 1998)
Figure 2 1 Production of grain legumes in the 12 European countries where the total harvest exceeded 100,000 t in 2013 Data from Eurostat (2016) (faba bean, pea, soybean, lupins) and FAOSTAT (2016) (lentil, chickpea, common bean)
Lima beans are native to Guatemala, Mexico, and Peru It spread throughout the Americas after being domesticated, and the Spaniards brought it to the Pacific Islands and the Philippines It then spread to South-East Asia, while the slave trade brought the Lima bean to Western and Central Africa The Lima bean has now become widely naturalized throughout the tropics It thrives in humid, subhumid, and semi-arid tropical regions, as well as warm temperate climes In humid regions, Lima beans are frequently intercropped with
6 cereal crops, root crops, or other crops, however in drier climes, it is grown as a single crop (Bosch et al., 2002)
Lima bean is a herbaceous plant with two distinct development patterns The perennial form is an indeterminate, robust, climbing and trailing plant that grows up to 2-6 m tall and only flowers axillarily It has 2 m long swollen and meaty roots Annual Lima bean is a bushy, pseudo-determinate plant that grows to be 0.3-0.9 m tall and has both terminal and axillary flowering The roots are slender The stems can grow to be up to 4.5-8 m long The leaves are alternating and trifoliate, with elliptical leaflets that measure 3-19.5 cm long and 1-11 cm wide The inflorescences are 15 cm long and produce 24 bisexual white or violet blooms (Bosch et al., 2002) Other names for butter beans are sieva beans, double beans, Madagascar beans, and wax beans Serrano-Serrano and colleagues (2012) diskovered that during the cultivation process, people in this region domesticated this plant and produced two types of seeds: Large-seeds (Lima) (in the West Andes) and Small-seeds (Sieva) (in Central America) (Serrano‐Serrano et al., 2012) The giant grain was domesticated circa
2000 BC, while the little grain was domesticated in Mesoamerica around 800 AD (Motta‐ Aldana et al., 2010) This species of bean has a high nutritious content and thrives and develops well in the cLimate of our nation However, it only appears intermittently in recipes such as bean soup and stews, and there are few items created from beans in the globe, with canned or frozen beans being the most acceptable That is why it is beneficial to do study in order to diskover and maximize its full potential
Bean cultivars currently differ significantly in terms of the number of days from planting to harvest, size, and shell color Bean varieties are then classified into three categories: Baby- seed Lima bean, Fordhok-seed Lima bean, and speckled Lima bean The Baby seed group includes varieties such as Eastland and Parkers; the Fordhook seed group includes popular types such as Carolina Sieva, Jersey, and Bliss; and the speckled seed group includes varieties such as Christmas and Jackson Wonders (Glancey et al., 1997), (Bailey 1896) We use protein from the Christmas variety in our report
Table 2 1 Description of commercially available Lima bean varieties
Maturi ty-days to harvest
Its small size and tendency to set pods on the ends of raceme can reduce harvest recovery with pod-stripper combines
Eastland Baby 82 A,B,C,D Several Closely related to M-15 Has a more upright plant
8 architecture, more conducive to harvest by pod-stripper combine
Larger plant type that sets more pods in the top of the plant A good yielding variety
Wonder Speckled 85 None Several Speckled bean type, with more heat resistance than others
New variety that has demonstrated good yield potential in trials
Has demonstrated good yield potential in later plantings in Delaware when conditions are cooler
Yields equal to F1072 Has a larger berry size
Has outyielded F1072 in trials Recommended for trial use
Proximate composition of the Lima bean samples
Table 1 shows the results of the proximate composition of the Lima bean samples All of the Lima bean samples had low moisture content and fell within the acceptable range of
0 - 13.5% (James 2013) The moisture level of the samples ranges from 7.01% for roasted Lima beans to 8.05% for fermented Lima beans The moisture content of a food is an indicator of its water activity; foods with high moisture content perish quickly owing to microbial activity (Onyeike et al., 1995) The protein content of Lima bean samples ranged from 19.79% in roasted Lima beans to 22.24% in raw Lima beans The protein content of raw Lima beans was lowered by all processing techniques Similar results on protein content reduction after processing such as cooking and roasting have been reported (Ndidi et al., 2014) Low fat levels were seen in both the processed and raw Lima beans, with the raw beans having a fat content of 1.20% and the roasted beans having 1.05% There have been reports that Lima beans are low in fat (Handbook 1986) The beans' low fat content is advantageous since it lowers the risk of heart attacks and raises blood cholesterol levels (Hession et al., 2009) Additionally, processing decreased the Lima beans' crude fibre content Crude fibre content in the germinated sample was greatest (6.22%) across all processing procedures The Lima bean seeds were dehulled after soaking, which is likely why roasting and fermentation were very low (p 8 > 6 > 2 > 4, respectively The greatest emulsifying ability is at pH 10 with
54 emulsifying activity of 130.44 m 2 /g of PCS10 and 103.48 m 2 /g of SPI sample, similar to the ESI of the two samples (199.44 and 191.04 minute, respectively), this consistent with research of (Tirgar et al., 2017) At high pH range (alkaline pH), the protein structure will expand, leading to exposure of hydrophobic groups as well as protein disintegration, which will facilitate the formation of hydrophilic and lipophilic interactions between the water phase and the fat phase helps create more emulsions Meanwhile, the emulsifying ability at pH 4 was recorded as the lowest in both PCS10 and SPI samples (6.45 and 6.72 m 2 /g, respectively) Because this is the pH range close to the protein's isoelectric point where solubility and protein–protein interactions are greatest (Garba et al., 2014) Emulsion systems at other pH values will be more stable than at isoelectric pH At pH values far from the isoelectric point, because the droplets are dispersed farther apart, interactions between proteins adsorbed to other droplets occur weakened, which may promote stronger interactions between proteins adsorbed into the same droplet to form a strong film on the surface Having a large protein–protein binding force will disrupt the balance between hydrophilicity and hydrophobicity of the protein (McClements & science 2004)
Furthermore, the hydrophobic portions of the protein molecule are adsorbed on the droplet surface when oil droplets form in PCS10 solution, whilst the hydrophilic portions— such as the carboxyl groups (-COOH) and amino groups (-NH2)—are in the aqueous medium At pH>pI, the (-COOH) group transforms into (-COO-), while at pH