C H A P T E R

2
Agricultural-Based Protein
By-Products: Characterization
and Applications
G.S. Dhillon*, S. Kaur**, H.S. Oberoi†,
M.R. Spier‡, S.K. Brar§
*Department of Agricultural, Food and Nutritional Sciences (AFNS), University of
Alberta, Edmonton, AB, Canada; **Department of Biological Sciences, University of
Lethbridge, AB, Canada; †Department of Post-Harvest Technology division, Indian
Institute of Horticultural Research (IIHR), Bangalore, India; ‡Federal University of
Paraná, UFPR, Post-Graduation in Food Engineering, Curitiba, Brazil; §INRS, ETE,
University of Quebec, QC, Canada

1 INTRODUCTION additives, anticarcinogenic compounds, dietary

fiber, biofuels, single-cell proteins, organic acid-
The abundant quantity of agro-industrial fermented beverages, compost, and biodegrad-
wastes and by-products are produced both in able plastics.
the organized and unorganized sectors through- Recently, the growing awareness of nutrition,
out the world. Most of the time, these valu- health, and environmental consciousness of con-
able resources are not efficiently used or find sumers is compelling researchers to search for
low-value applications, such as animal feed or sources of good quality sustainable plant proteins
soil composting. These waste feedstocks or by- for food applications. With the increased industri-
products can be efficiently used for the extrac- al demand of protein sources in the food industry,
tion and production of high-value-added prod- there has been a recent upsurge of research efforts
ucts, such as amino acids, bioactive peptides, to extract dietary proteins from plant- and animal-
antimicrobials, enzymes, edible oils, essential based waste feedstocks or by-products. Besides
oils, polyphenolic compounds, pigments, food food applications, proteinaceous feedstocks find

Protein Byproducts
http://dx.doi.org/10.1016/B978-0-12-802391-4.00002-1
21 Copyright © 2016 Elsevier Inc. All rights reserved.
22 2.  Agricultural-Based Protein By-Products: Characterization and Applications

various other applications, such as biopolymers, profile/CVF15). The production of corn is ex-
biocomposites, bioplastics, wastewater treatment, pected to increase because of its utilization for
and agriculture. bioethanol production. Distiller’s grain, which
The potential agricultural-based sources for is a by-product of bioethanol manufacturing,
the extraction of proteins are fruit- and veg- mainly from corn and wheat, comes in different
etable-processing industries, distilleries, and forms. Lipids and carbohydrates are used for
oilseed by-products. Agricultural produce pro- biofuels production, and the leftover fraction,
cessing results in an abundant quantity of by- distiller’s grain, is rich in protein content. Protein
products, including protein-rich by-products. is the second most abundant component of corn
By-products resulting from the production of after starch. The protein content of different corn
bioenergy are also generally rich in proteins, varieties ranges from 6% to 12% on a dry basis.
because of the utilization of carbohydrate and Wine is an alcoholic beverage made from
lipid fractions. These sustainable low-cost and fermented grapes or other fruits, such as pome-
abundant protein by-products can be consid- granates, berries, and apples. In 2014, the total
ered as feedstocks for various applications, such worldwide production was 28,230,400 (liters
as food formulations, biomedicine, bioplastics, per thousand) (Teixeira et al., 2014). The top
biopolymers, among others. This chapter mainly four wine producers are France (16.54%), Italy
focuses on the plant- and animal-based protein (15.85%), Spain (13.53%), and the United States
by-products, their physicochemical and biologi- (10.77%) (www.wineinstitute.org/resources/
cal characterization, and finally their high-value statistics). The wine-making process generates a
applications. large amount of solid waste (up to 30% w/w of
the material used), mainly consisting of organic
wastes, namely fruit stems, skins, and seeds.
2  PLANT-DERIVED PROTEIN
Bioactive compounds from winery by-products
BY-PRODUCTS
have disclosed interesting health-promoting
activities both in vitro and in vivo (Teixeira
2.1  By-Products from Bioenergy et al., 2014). The by-products resulting from vi-
Production, Breweries, and Wineries nification contain appreciable amounts of pro-
Because of the quick depletion of nonrenew- tein, which currently do not find any high-value
able fossil fuel reserves, biofuel production is in- applications. Moreover, the type of by-products
creasing at a fast pace. Biofuels are generally pro- produced during wine making is closely de-
duced from renewable agricultural feedstocks. pendent on the specific vinification procedures,
Hence, by-products resulting from bioenergy which also affect the physicochemical properties
(eg, bioethanol and biodiesel production) and of the by-products, the characteristics of which
breweries and wineries are produced in abun- determine its further use and specific valoriza-
dant quantities worldwide. These by-products tion path in which it could be integrated.
are considered a rich source of proteins. Similarly, biodiesel is produced from Jatro-
Corn (Zea mays L.) is one of the most impor- pha oil, melon, palm oil, soybean, rapeseed oil,
tant food and industrial crops in the United sunflower oil, and used oil can be used to make
States. It is widely used for the production of biodiesel. Jatropha is very important in the
bioethanol. In the 2014-15 marketing year, world production of biodiesel because it is a noned-
annual production of corn was around 988.077 ible plant and hence does not compete with the
million metric tons (MMT), of which the US food oils. The solid by-products obtained after
alone contributed 36.5% (http://www.barchart. oil extraction from seeds are known as oil cakes
com/commodityfutures/Tocom_Corn_Futures/ or oil meal. Their composition varies widely

1.  General introduction

 2  Plant-derived protein by-products 23
TABLE 2.1 The Composition of Oil Cakes Originating 2.2  Oil Crops
from Different Types of Plant Sources
The amino acid content of different agricul-
Protein
tural by-products is provided in Table 2.2.
Oil cake Dry matter (%) content (%)
Canola protein (CP): Canola or rapeseed (Bras-
Canola oil cake 90.0 33.9 sica napus) is an important oilseed crop in many
Coconut oil cake 88.8 25.2 countries and is the second most abundant
Cottonseed cake 94.3 40.3 source of edible oil in the world. Canola seeds
contain approximately 40% oil and 17–26% pro-
Groundnut oil cake 92.6 49.5
tein (Uppstrom, 1995). Canola meal (CM), the
Mustard oil cake 89.8 38.5
by-product of canola oil extraction, is highly
Olive oil cake 85.2 6.3 proteinaceous and contains up to 50% protein
Palm kernel cake 90.8 18.6 on a dry basis. The main protein constituents of
Sesame oil cake 83.2 35.6 CM are napin and cruciferin, the storage pro-
Soy bean cake 84.8 47.5 teins, and oleosin, a structural protein associ-
ated with the oil fraction (Uppstrom, 1995). CP
Sunflower oil cake 91.0 34.1
is currently used for human food and animal
Adapted from Kolesarova et al. (2011). feed. However, because of its abundance, CM
can be exploited for other nonfood applications,
depending on the quality of seeds or nuts, grow- including bioflocculation.
ing conditions, and extraction methods. Oil Soy protein (SP): This protein is produced from
cakes can be either edible or nonedible. Edible soybeans (Glycine max) by a multistep process
cakes have a high protein content ranging from that removes the oil and indigestible compo-
15% to 50%. The compositions of oil cakes origi- nents. Depending on the processing steps used,
nating from different types of plants are listed SP ingredients may take the form of isolated soy
in Table 2.1. The protein content in different oil protein (ISP), SP concentrate, or soy flour. Cur-
cakes ranges between 6.3% and 49.5%. Oil cakes rently, SP find applications in human food and an-
are currently used mainly for feed applications imal feed (Hertrampf and Piedad-Pascual, 2000;
to poultry, ruminant, and in the fish and swine Montgomery, 2010). It is a potential raw material
industries (Franke et al., 2009; Molina-Alcaide for SP-based bioflocculants. Recent studies dem-
and Yanez-Ruiz, 2008; Mushtaq et al., 2009; onstrated the ability of SP-based bioflocculants
Soren and Sastry, 2009). Some of them are con- for the treatment of diatomite, kaolin, and kanto
sidered to be suitable organic nitrogenous fertil- loam (Liu et al., 2012; Piazza and Garcia, 2010a;
izers. Several cakes have been used for produc- Seki et al., 2010). Considering the abundant pro-
tion of proteins, enzymes, antibiotics, vitamins, duction of SP, various other high-value applica-
antioxidants, mushrooms, and ethanol (Bernes- tions can also be developed.
son, 2007; Cervero et al., 2010; Soren and Sas-
try, 2009; Vastag et al., 2011).
The by-products resulting from bioetha-
2.3  Cereal Proteins
nol, biodiesel, and breweries and wineries are
widely used as an animal and aquaculture feed Barley proteins: Barley grain (Hordeum vulgare
(Traub, 2014). Further development of the high- L.) is the fourth most widely grown cereal in
value applications of the by-products resulting the world next to wheat, rice, and corn (Yalçin
from biofuels production will enhance the eco- et al., 2008). In 2009 and 2010, global production of
nomic viability of the overall biofuel production barley was 152 and 124 MMT, respectively (FAO-
process. STAT, 2010). In Canada, nearly 80% of the barley

1.  General introduction

 TABLE 2.2 Amino Acid Composition of Different Proteins (g Amino Acid: 100 g Unless Otherwise Mentioned)

24
Canola
(Brassica Soy protein Rice bran Barley protein Tomato Seeds
Zein napus, cv. isolate protein Casein fractions PGF/PF protein (mg/g
(Mossé, 1961; Altex) (Tzeng (Wang (RBP) (Wang (Wang proteins (%) (Wang protein) (Sarkar
Class Amino acid Pomes, 1971) et al., 1988a, b) et al., 2008) et al., 1999) et al., 2010) et al., 2010) and Kaul, 2014)

2.  Agricultural-Based Protein By-Products: Characterization and Applications

Basic Lysine 0 6.60 5.23 ± 0.01 5.5 7.10 1.81/5.03 59.63
Arginine 1.8–4.71 7.28 7.35 ± 0.35 9.0 3.30 3.91/4.14 —
Histidine 1.1–1.32 3.18 2.81 ± 0.24 3.0 2.70 2.32/1.81 25.01
Acidic Aspartic acid (as 4.61 (4.5) 7.79a 11.47 ± 0.71a 10.5a 6.30a (4.34/9.05) —
asparagine)
Glutamic acid 1.5–26.9 20.81b 20.67 ± 0.83b 15.3b 19.0b (26.74/15.15) —
(as glutamine) (21.4)
1.  General introduction

–OH Serine 5.7–7.05 4.41 5.32 ± 0.09 5.3 4.60 5.23/5.02 —

Threonine 2.7–3.45 4.81 3.98 ± 0.13 4.4 3.70 3.50/4.52 36.49
Tyrosine 5.1–5.25 3.19 3.61 ± 0.18 3.7 5.50 3.46/2.56 —
–S Methionine 2.0–2.41 2.24 0.92 ± 0.07 2.0 2.60 1.56/1.70 —
Cysteine 0.8–0.83 2.08 0.05 ± 0 2.6 0.04 1.33/0.31 —
Nonpolar Glycine 0–0.7 4.60 3.74 ± 0.27 6.1 1.60 3.54/8.31 —
Alanine 8.3–10.52 4.53 3.72 ± 0.06 6.8 2.70 4.52/9.29 —
Valine 3.1–3.98 5.65 4.28 ± 0.32 5.7 6.00 5.61/8.03 55.19
Leucine 19.3–21.1 7.47 6.79 ± 0.83 8.0 8.40 7.56/7.22 77.90
isoleucine 5–6.2 4.47 4.35 ± 0.11 3.0 4.90 3.46/3.05 49.30
Phenylalanine 6.8–7.3 4.67 5.14 ± 0.58 5.1 4.50 4.54/3.16
Tryptophan 0.16 ND ND 7.0 ND ND/3.56 12.36
Proline 9.0–10.53 6.22 5.13 ± 0.33 ND ND 16.57/8.09 —
SSA — — — — — — 30.58
AAA — — — — — — 87.32
AAA, aromatic-containing amino acids; ND, not detectable; PGF, pearled grain flour; PF, pearling flour; SSA, sulfur-containing amino acids.
a
Asparate + asparagine.
b
Glutamate + glutamine.
 2  Plant-derived protein by-products 25
crop is used as livestock feed, 15% for malting, generally used as animal feed. Limited research
and <5% for direct human intake. Recently, there has been carried out to evaluate barley protein
has been an increasing interest in barley as a food fractions, focused mainly on hordein or whole-
ingredient, which is attributed to the health ben- grain proteins (Bamforth and Milani, 2004; Bilgi
efits of barley β-glucan, lignans, calcium, and and Çelik, 2004; Kapp and Bamforth, 2002).The
selenium. β-glucan, a soluble dietary fiber com- thorough research to identify value-added ap-
ponent, reduces blood cholesterol and glycemic plications for barley processing by-products is
index (Behall et al., 2006; Keenan et al., 2007). desirable. This will facilitate the expanded pros-
The approval of a health claim for soluble barley pects for barley protein fractions in food and
β-glucan by the US Food and Drug Administra- nonfood applications.
tion (FDA) further increased research efforts for The solubility profile of hordein indicates
barley food product development (FDA, 2005). that it has low solubility at pH 3–8 with the
After β-glucan isolation from barley grains, the least value attained at pH 5. High solubility was
remaining portions are a good source of proteins, observed at pH 10. A similar solubility profile
starch, and lipids (Baik and Ullrich, 2008). Cur- was observed for the glutelin and PGF protein
rently, there is no commercial application of these fractions, with the minimum values obtained at
fractions. Therefore, research in this direction is pH 5. An increase in protein solubility was also
sought to develop their full value. observed at pH 10 (Wang et al., 2010). The pH
The total barley grain protein composition profile indicates that these proteins are suitable
varies from 8% to 13% w/w, with various barley for their application as flocculants for the treat-
grain tissues enriched with specific protein types ment of wastewater streams having pH above
at different levels (Pomeranz and Shands, 1974). neutral; for example, oil sand tailings with pH
The major proteins in barley endosperm are hor- levels <8.5 (Table 2.3).
dein (35–45%) and glutelin (35–45%), whereas Zein: It is the major storage protein of corn
barley bran and germ are enriched in cytoplas- and comprises 45–50% of the total protein in
mic proteins (mainly albumin and globulin) corn. Currently, zein production is nearly 500
(Lâsztity, 1984). These protein fractions may tons/year from corn gluten meal. Zein belongs
display different functional qualities for vari- to the characteristic class of proteins known as
ous applications owing to different molecular prolamines, which occur specifically in cereals
structures. Generally, barley bran is removed (corresponding to hordein in barley and gliadin
from barley grains to yield a bright white kernel in wheat). Zein is not suitable for human intake
for various food applications through a barley owing to its negative nitrogen balance and poor
milling process known as pearling. The removed solubility in water. The other main disadvan-
grain layers are called the pearling flour (PF), and tage to utilization of zein in human foodstuffs
the rest is called the pearled grain floor (PGF). The is their poor “functionality.” Functional prop-
PF is a rich source of albumin and globulin pro- erties refer to those attributes that provide the
teins (Yeung and Vasanthan, 2001). However, it desired physical or sensory properties to the
is an underused and inexpensive by-product. food. For example, proteins do not only con-
In brewing industries, barley endosperm tribute nutritional quality to a food, but also
proteins (hordein and glutelin) are considered contribute textural properties, such as gelation
undesirable compounds or even contaminants (essential in meats, cheeses, gels, and so forth),
and are normally precipitated out in the barley adhesion (meats, bakery, pasta), emulsification
spent grain (BSG). Hordein and glutelin proteins (deli meats, soups, cakes), foaming or whipping
(20–30% w/w) in BSG are almost three times that (cakes, frozen desserts), and moisture sorption
of the whole grain (Jadhav et al., 1998). BSG is (intermediate moisture, shelf-stable foods).

1.  General introduction

26 2.  Agricultural-Based Protein By-Products: Characterization and Applications

TABLE 2.3 Characteristics of Protein By-Products Derived from Different Agricultural Sources

Protein Isoelectric pH Remarks Reference

Albumin 4.7 Used for commercial applications Ge et al. (1998)

Barley proteins 5 Barley bran and germ are rich in cytoplasmic Wang et al. (2010)
proteins (albumin and globulin)
Barley floor contains endosperm proteins
(hordein and glutelin.
No commercial application
High solubility at pH 10 makes it ideal for treat-
ment of wastewaters with pH above neutral
Canola meal 4.5–5.5 (best 5) High production of canola
protein
Casein 4.6 Chen et al. (1987)
Collagen 6 Maslennikova et al. (2013)
Corn zein 6.2 (varies between 5 and 9) Insoluble in water Shukla and Cheryan (2001)
Gelatin 4.8 Used for commercial applications Maslennikova et al. (2013)
Gluten Nearly 6 Wang et al. (2006)
Hordeins 6.5–8 Faulks et al. (1981)
Keratin 5.6–6 Cooper and Sun (1986)
Rice proteins 4.4 No current applications. Rice polishing pro- Watanabe et al. (2011)
duces large quantities of wastewater rich
in proteins and starch
Soya protein 4.5–5.5 Seki et al. (2010)

Zein is soluble in concentrated ethanol, high residues and paucity in basic and acid amino
concentrations of urea, high concentrations of acids accounts for the solubility behavior of
alkali (pH 11 or higher) or anionic detergents. zein. In whole corn, zein occurs as a heteroge-
Currently, zein is used for making fiber, adhe- neous mixture of disulfide-linked aggregates
sive, coating, ceramic, ink, cosmetic, textile, with a average molecular weight of 44,000 Da
chewing gum, and biodegradable plastics. (Pomes, 1971). The high isoelectric pH of zein
These new applications of zein appear prom- (pH 5–9), indicates its potential as a flocculating
ising, but require the development of low-cost agent to be used in wastewater treatments hav-
manufacturing methods. Currently, zein prices ing pH levels higher than neutral. However, the
vary from US$10 to $40 per kg, depending on poor solubility of zein restricts its use in various
purity, representing a very high-value addition industrial applications. Hence, different modifi-
to corn. However, it also confines more wide- cation techniques are sought to increase its solu-
spread applications of zein. This mandates find- bility and extend its applications in various sec-
ing practical ways to significantly lower the cost tors, such as food, bioplastics, and biopolymers,
of manufacturing and to increase the zein use. among others.
Zein is mainly rich in glutamic acid (21–26%), Gluten: It is the main structural protein com-
leucine (20%), proline (10%), and alanine (10%), posite of wheat and other cereals, including
but deficient in basic and acidic amino acids rye and barley. The protein fraction of gluten
(Table 2.2). High content of nonpolar amino acid comprises gliadins and glutenins, with gliadins

1.  General introduction

 2  Plant-derived protein by-products 27
containing monomeric proteins and gluten- increase in fruit processing has been generat-
ins containing aggregated proteins. Recently, ing millions of tons of agro-industrial wastes
Sapone et al. (2012) thoroughly reviewed the (both solid and liquid by-product) worldwide.
gluten-related disorders. Besides the well- These by-products are generally rich sources of
known disorder of celiac disease (CD), there are proteins and other bioactive compounds. Crude
spectrum of other acknowledged gluten-related protein composition (percent DM basis) of veg-
disorders. Hence, the number of individuals etable and fruit wastes are provided in Table 2.4.
preferring a gluten-free diet is much higher than Grapes (Vitis vinifera L.): Currently, up to 210
the projected number of CD patients, fueling a million tons of grapes are produced annually,
global market of gluten-free products reaching with 15% of the produced grapes produced for
US$2.5 billion in worldwide sales in 2010. This the wine-making industry. Grapes are consid-
trend is supported by the concept that along ered as a rich source of bioactive compounds.
with CD, other conditions related to the inges- The major by-products from wine-making ac-
tion of gluten have emerged as health concerns tivity are organic wastes (grape pomace, con-
among many individuals in the world, which taining seeds, pulp and skins, grape stems, and
has caused emerging health care concerns and grape leaves), wastewater, and inorganic wastes
reduced its market potential as food ingredient. (diatomaceous earth, bentonite clay, and perlite)
Because of consumer preference for gluten-free (Oliveira et al., 2013).
diets, gluten has become a significant by-prod- Grape pomace is produced during the pro-
uct of grain production. Hence, new applica- duction of must (grape juice) by pressing whole
tions of gluten should be explored. grapes. Currently, 9 million tons of grape pom-
Rice proteins: Rice, a cereal grain, is the most ace are produced per year in the world, which
widely consumed staple food for a large part constitutes 20–30% w/w on average of the to-
of the world’s human population, especially in tal grapes used for wine production (Dwyer
Asia (Vaughan, 1994). The demand for polished et al., 2014; Kataliníc et al., 2010). The moisture
rice, so-called “rinse-free rice,” has increased. percentage of grape pomace varies from 50% to
As a result, rice manufacturing plants produce 72% depending on the grape variety considered
a large amount of rice washing drainage, which and its ripening state. The insoluble residues
is a water pollution source. This drainage con- from this material have a lignin content ranging
tains a relatively high amount of solid particles from 16.8% to 24.2% and a protein content lower
composed of starch and proteins (a by-product than 4%. The main polymer-type constituent of
consisting of the inner bran layers of the kernel the cell walls present in grape pomace are peptic
with part of the germ and a small portion of substances, ranging from 37% to 54% of cell wall
the starchy interior) produced during rice pol- polysaccharides. Cellulose is the second type of
ishing (Watanabe et al., 2006, 2009). There is no cell wall polysaccharides in abundance in grape
practical application of this protein-rich waste. pomace, varying from 27% to 37% (González-
Economical processes can be developed for the Centeno et al., 2010). However, this high content
extraction of proteins and starch for various ap- of nondigestible polysaccharides results in gas-
plications. trointestinal disturbances in animals. Therefore
additional fermentation processes may be re-
quired for its valorization to animal feed.
2.4  Fruit Wastes and By-Products Considering the separate fractions of grape
Agro-based industries, especially apple-pro- pomace (seeds and peels), the relative propor-
cessing industries, are experiencing a surge in tion of seeds ranges from 15% to 52% of the dry
their growth around the globe. This enormous material (Ghafoor et al., 2009; Nawaz et al., 2006;

1.  General introduction

28 2.  Agricultural-Based Protein By-Products: Characterization and Applications

TABLE 2.4 Crude Protein Composition (Percent DM Basis) of Vegetable and Fruit Wastes
Scientific name Crude protein (CP)

FRUIT WASTES

Banana peels (Wadhwa and Bakshi, 2013) Musa acuminata 8.1

Muskmelon peels (Wadhwa and Bakshi, 2013) Cucumis melo 9.5
Watermelon peels (Wadhwa and Bakshi, 2013) Citrullus lanatus 7.9
Citrus pulp (without peels) (Wadhwa and Bakshi, 2013) Citrus limetta 10.5
Apple pomace (NRC, 2001) Malus domestica 7.7
Grape pomace (Zalikaranab et al., 2007) (Vitis vinifera L.) <4%

VEGETABLE WASTES

Sugar beet leaves (Wadhwa and Bakshi, 2005) Beta vulgaris 21.9

Cauliflower leaves (Wadhwa and Bakshi, 2005) Brassica oleracea B. 17.0
Cabbage leaves (Wadhwa and Bakshi, 2005) Brassica oleracea C. 19.9
Black chick pea plant (Wadhwa and Bakshi, 2005) Cicer arietinum 13.6
Pea vines (Wadhwa and Bakshi, 2005) Pisum sativum 11.8
Radish leaves (Wadhwa and Bakshi, 2005) Raphanus sativus 19.4
Summer squash vines (Wadhwa and Bakshi, 2005) Cucurbita pepo L. 13.9
Baby corn husk (Bakshi and Wadhwa, 2012a) Zea mays Linn. 11.6
Carrot (NRC, 1989) Daucus carota 9.9
Potato (NRC, 1989) Solanum tuberosum L. 9.5
Snow peas (Bakshi and Wadhwa, 2012b) Pisum sativum var. saccharatum 23.2
Pea pods (Wadhwa and Bakshi, 2013) Pisum sativum 9.8
Carrot pulp (Wadhwa and Bakshi, 2013) Daucus carota 7.2
Bottle gourd pulp (Wadhwa and Bakshi, 2013) Lagenaria siceraria 24.3
Sugar beet pulp (NRC, 2001) Beta vulgaris 10.0
Tomato pomace (Wadhwa and Bakshi, 2013) Solanum lycopersicum 22.1
Adapted from Wadhwa and Bakshi (2013)

Stamatina et al., 1995). The composition of grape the extraction method used. The grape pomace
seeds (w/w) consists of up to 40% fiber, 16% es- and stems are currently not valued as highly
sential oil, 11% protein, 7% complex phenolic profitable waste and are mainly directed to com-
compounds, like tannins, and other substances posting or discarded in open areas, potentially
like sugars and minerals (Campos et al., 2008). causing environmental problems.
Likewise, grape skins constitute 65% of the to- Apple (Malus domestica): According to the Food
tal material of grape pomace on average. Grape and Agriculture Organization of the United
skin has been reported as a rich source of pheno- Nations (FAO), worldwide apple production
lic compounds, even though the final yield is de- exceeded 69,603,640 tons in 2008–09 (http://
pendent on the specific vinification process and faostat.fao.org). The processing of apples to

1.  General introduction

 2  Plant-derived protein by-products 29
various products yields millions of tons of waste, Pineapple (Ananas comosus): The posthar-
for example, 25–30% solid pomace waste and vest processing of pineapple fruit yields skins,
5–10% liquid sludge (Dhillon et al., 2013). Apple crowns, and waste from fresh trimmings and the
pomace and apple pomace sludge resulting from pomace after extracting the juice. Raw pineapple
apple processing contains 2.9–5.7% protein (dry waste (on a DM basis) contains 4–8% CP. Banana
basis), which amounts to 28.8–33.8 g/L protein is another common fruit consumed in abundant
(Dhillon et al., 2011, 2013). And the apple waste quantities throughout the world. Banana leaves,
also contains cull fruits (fresh fruits unsuitable pseudo-stems, and ripe banana peels contain
for human consumption). about 10–17%, 3–5%, and 8% CP, respectively.
Citrus pulp: Pulp from citrus processing Finally, there are various other fruits that con-
(50–70% of the fruit by weight) is the residue tribute abundant quantities of by-products dur-
left after extraction of the juice. It contains ing postharvest and processing and hence can
60–65% peel, 30–35% internal tissues, and up be viewed as a potential source for proteins and
to 10% seeds (Crawshaw, 2004). Citrus pulp is other value-added products.
usually made from oranges (60%), grapefruits,
and lemons. It contains 5–10% CP and various
other nutritional elements. Citrus molasses is a 2.5  Vegetable-Processing Wastes
by-product obtained after citrus juice extraction. Baby corn (Zea mays Linn.): Thailand is the ma-
The fresh pulp mixed with lime is pressed to re- jor baby corn producing and exporting country
move moisture. The resulting liquid (press juice) in the world. It is eaten both raw and cooked and
is screened to remove the larger particles, ster- is used as a delicacy in Asian cuisine. Only 15%
ilized, and concentrated. It is a thick, viscous, is the edible baby corncob. The other 85% con-
dark-brown to almost black liquid and is known stitutes the outer peels with a silky threadlike
as citrus molasses. The composition of citrus mo- structure called baby corn husk, which is consid-
lasses (60–65% sugars and 4–5% CP) is equiva- ered a waste material and a source of environ-
lent to sugarcane molasses. mental pollution (Bakshi and Wadhwa, 2012a).
Mango: This fruit, belonging to the genus Man- Other by-products are tassels and green plant
gifera, is one of the most important fruits marketed material.
worldwide with a global production exceeding Carrot (Daucus carota): Carrots are widely
26 million tons in 2004 (FAOSTAT, 2004). It is cul- used as vegetables around the world. The post-
tivated or grown naturally in more than 90 coun- harvesting and processing of carrot results in the
tries worldwide (mainly tropical and subtropical production of various by-products: cull (grade-
regions) and is known to be the second largest out), carrot tops, and carrot pomace after juice
tropical fruit crop produced in the world (Joseph extraction. Carrot by-products find limited ap-
and Abolaji, 1997). Mango processing generates plications as an animal feed.
approximately 50% of the fruit handled as waste. Peas (Pisum sativum L.): Next to soybeans,
The edible pulp makes up 33–85% of the fresh groundnuts, and beans, peas are one of the most
fruit, while the peel and the kernel amount to important legume crops. After harvesting sweet
7–24% and 9–40%, respectively, on a fresh weight peas, which are eaten as a vegetable, the leftover
basis (Wu et al., 1993). After processing mango, plant is called pea vine, which contains around
the available by-products and waste includes 11.8% CP. Pea vine protein fractionation shows that
cull fruits (fresh fruits unsuitable for human con- it contains the highest concentration of albumins,
sumption), mango kernel meal, de-oiled mango followed by glutelins, globulins, and prolamins.
kernel meal, and mango peels. The mango waste After harvesting sweet peas, the mature plant is
contains 10.3% CP (Sunita and Rao, 2003). left in the field for drying and is called pea straw.

1.  General introduction

30 2.  Agricultural-Based Protein By-Products: Characterization and Applications

It contains 5–10% CP. With higher protein content Some potato species are known to have high
and less fiber, pea straw has a higher nutritive protein content in their tubers. Among the
animal feed value than cereal straws. After shell- health-promoting compounds are carotenoids,
ing peas, the leftover material is empty pea pods, flavonoids, caffeic acid, and the tuber stor-
which contain high CP concentrations—19.8%. age protein patatin, all of which exhibit activ-
Snow peas (Pisum sativum var. saccharatum): A ity against free radicals and have antimicrobial,
variety of pea, but unlike sweet peas, these are antiinflammatory, and antiallergic properties
valued for their pods rather than just the beans (Brown et al., 2007; Ritter et al., 2008). During the
inside and are eaten whole with the pod. Snow peak production season, it is a problem for farm-
peas are delicate and sweet in flavor. Frost-af- ers to dispose of the surplus and cull potatoes
fected snow peas are considered unfit for human because of strict environmental laws. The only
consumption, fail the quality control test, and are option for the farmers is to feed them to the live-
not exported. Cull snow peas contain high CP stock. However, raw potatoes are not very pal-
concentration of 23–25% with 35.8% total sugars atable and have a laxative effect and, therefore,
on DM basis. These are an excellent source of not considered good for animal feed.
vitamins A, B complex, C, and K. Also, they are Sweet potato: Sweet potatoes are used as vegeta-
rich in pigments, such as lutein and zeaxanthin, bles throughout the world. Shochu is a traditional
which help promote vision. An integrated pro- Japanese liquor made from rice, sweet potatoes,
cess for extraction of protein and other bioactive and barley. Sweet potatoes are widely used for
compounds will create extra income for growers. making this beverage and it is very popular in
Potato (Solanum tuberosum L.): Potatoes are the southern Japan. Recent increases in shochu pro-
most popular vegetable throughout the world. duction have resulted in an abundant quantity of
Along with wheat and rice, potato ranks as one distillery by-products. A small fraction of these
of the most important staple crops in the human by-products have been used as an animal feed
diet around the globe. Potatoes contain various (Mahfudz et al., 1996) and the remaining portion
compounds with high bioactivity, especially the have been discarded into the ocean. This practise
potato skin. On the first cell layers under the of by-product management becomes problematic
skin are rich sources of various bioactive com- from the perspective of environmental protection,
pounds. Besides starch as main component, and it mandates optimal treatment of distillery by-
potato tubers also contain an ample quantity products, which is also important for the success
of bioactive compounds, such as proteins, pep- of commercial shochu production. Hence, devel-
tides, carotenoids, polyamines, polyphenols, oping new applications for the shochu distillery
suberins, glycoalkaloids, and dietary fiber. The by-products (SDBs) originating from the sweet
fresh potatoes contain 65–75% starch (depend- potato is necessary from the standpoint both of
ing on the variety) and 9.5% CP and other com- the economy and of environmental health.
pounds. Most of them enhance human health SDBs contain abundant amounts of health-
and hence find potential applications in food related food ingredients, such as essential amino
industry. Potato processing, generally caused acids, vitamin E, dietary fiber, citric acid, yeast,
by potato-based fast-food products (chips and and minerals. The filter cake of the sweet pota-
fries), results in the production of by-products, to–fermentation residue has potential as a food
mainly peel waste which is generally used for material, owing to its high CP content and an
animal feed or soil improvement. Approximate- amino acid balance that is superior to that of ce-
ly 40% of potatoes are wasted, representing ap- real grains (Wu and Bagby, 1987).
proximately 10 tons/day of residue (Barampou- Sugar beet (Beta vulgaris): More than 200,000
ti and Vlyssides, 2005). tons of red beet are produced in Western Europe

1.  General introduction

 3  Animal-derived protein by-products 31
annually, most of which (90%) is consumed as acid (Persia et al., 2003). Unlike many other
vegetable. The rest are processed into juice and plant proteins, tomato seed has also been re-
food colorant, the latter commonly known as ported to have a high lysine content (Brodowski
beetroot red (Henry, 1996). The pomace from and Geisman, 1980). The net protein retention of
the juice industry, accounting for 15–30% of the whole tomato seed meal, defatted tomato seed
raw material is used as an animal feed or com- meal, and tomato seed protein concentrate was
post (Otto and Sulc, 2001). The different wastes studied as 2.65, 2.52, and 2.51, respectively, as
and by-products of sugar beet include the leaves compared with 2.91 for casein (Sogi et al., 2005).
(22% CP) and pulp, which is mainly composed Nevertheless, underused tomato seeds—with
of pectin (28.7% DM basis); cellulose (20% DM high protein content and good nutritional attri-
basis); hemicellulose (17.5% DM basis); CP butes—can be a considered as a potential source
(9% DM basis); and lignin (4.4% DM basis) (Ja- of sustainable protein supplement for future
cob, 2009). The sugar beet pulp is dried and sold food formulations.
as dried sugar beet pulp or mixed with molasses
to form dried molasses beet pulp.
Tomato (Lycopersicon esculentum L.): Tomatoes
2.6  Value-Added Options for Fruit and
are grown worldwide with an annual produc- Vegetable Postharvest and Postprocessing
tion of nearly 100 million tons (Kalogeropoulos Wastes
et al., 2012). As per statistics from the World Pro- Fruit and vegetable wastes are highly fer-
cessing Tomato Council, more than 30 million mentable and perishable, mainly because of
tons of tomatoes are processed annually world- high moisture, total soluble sugars, and CP con-
wide to produce tomato juice, ketchup, canned tents. During the peak production or processing
tomatoes, sauces, paste, puree, powder, and season, massive quantities of these valuable re-
many other products (Zuorro et al., 2014). To- sources are available and cannot be consumed
mato processing simultaneously generates large at the same pace as they become available. Thus,
quantities of solid by-products, mainly peel and they become surplus and can cause environ-
seeds, usually raising environmental concerns. mental pollution. Therefore, suitable methods
Tomato waste is made up of culled tomatoes should be adopted to conserve such resources,
(14–20% CP) and tomato pomace (19–22% CP). which can be used for extraction of valuable
The culled fruits may be damaged, diseased, too components, such as proteins and other bio-
small, or misshapen and do not meet the grading products. This will also help mitigate environ-
standards for sale in the fresh market or for pro- ment pollution and create extra revenues.
cessing. Tomato pomace is a mixture of tomato
peels, seeds, and small amounts of pulp that re-
main after processing. Although, tomato peel is 3  ANIMAL-DERIVED PROTEIN
acknowledged to be high-value raw material for BY-PRODUCTS
lycopene, tomato seeds have attracted limited
attention. A recent study conducted by Sarkar Whey or Lactoserum: Whey (also known as lac-
and Kaul (2014) showed that tomato seeds are a toserum), the major dairy by-product through-
potential source of protein, having all amino ac- out the world, is readily available in most milk-
ids significantly higher than (WHO/FAO/UNU producing areas. It has around 5% dry matter,
2007) recommendations and a high calculated 12–13% CP (DM basis), and 60–70% lactose (DM
PER value of 2.66 (Table 2.2). Tomato seeds have basis). The protein content of whey varies from
been reported to contain approximately 24.5% 0.55–65% on dry weight basis. Recently, whey
CP and are highest in glutamic acid and aspartic has been used for the production of various

1.  General introduction

32 2.  Agricultural-Based Protein By-Products: Characterization and Applications

health products, such as whey protein concen- clay flocculation. Flocculation was completed
trate, lactalbumin, and whey protein hydro- within 24 h irrespective of the addition of cal-
lysates. Whey protein contains around 35% cium chloride. The same research group dem-
β-lactoglobulin, 12% α-lactoalbumin, glycomac- onstrated that beef-skin gelatin and hydrolyzed
ropeptide, ∼8% immunoglobulins, ∼5% serum fish collagen were found to improve flocculation
albumin, and ∼15% minor proteins. of clay suspension when pH 5.5 buffer was add-
Casein: This is the main protein component of ed (Piazza and Garcia, 2010a). However, gela-
milk. Casein is amphiphilic in nature. The iso- tin is costly as it finds commercial applications
electric point of casein is 4.6 (Table 2.3), above in food and other industries. The extraction of
which it is negatively charged and is soluble gelatin from waste sources using economical
in water (Chen et al., 1987). The conformation methods will shift toward feasible flocculation
of casein is almost similar to the denatured processes.
globular proteins. The high number of proline Albumin: Belonging to a family of globular
residues in casein cause particular bending of proteins, it is found in the blood plasma of hu-
the protein chain and inhibit the formation of mans, animals (eg, cattle, sheep, goats, horses,
close-packed, ordered secondary structures. Ca- chickens, and pigs, among others), and eggs. Se-
sein does not have any tertiary structure, which rum albumin is a protein found in small quanti-
describes its stability against heat denaturation, ties in the plasma phase of animal blood. Serum
as there is very little structure to unfold. Also, albumin has a wide range of commercial ap-
in the absence of tertiary structure, there is plications, depending on the grade, purity, and
substantial exposure of hydrophobic residues, quality of the product. Albumin from different
which results in strong association reactions of sources, especially bovine serum albumin, has
casein and renders them partially soluble in wa- found potential applications in medical and bio-
ter. The major use of casein has been as a food logical laboratories. It has been widely used in
ingredient to augment physical properties, such various other applications, such as diagnostics,
as foaming and whipping, thickening and water biotechnology, aquaculture, food and beverages,
binding, emulsification, and texture. Casein also chemicals and enzymes, and others.
enhances food nutrition value. Further, casein is Recent studies demonstrated the flocculation
used as wood adhesive, for leather finishing and potential of albumin (Seki and Suzuki, 2003;
paper coating, and in synthetic fibers, as well Seki et al., 2009). Seki and Suzuki (2003) carried
as plastics for buckles and buttons. However, out the flocculation of diatomite by methylated
the flocculation potential of casein has not been egg albumin. Diatomite is composed of approx-
much explored. Few studies demonstrated the imately 90% SiO2 and it has a negative charge
ability of casein as flocculent (Seki et al., 2004). above pH 3. Egg albumin has an isoelectric
Gelatin: A heterogeneous mixture of water-sol- point between pH 4 and 5 and is also negatively
uble proteins of high average molecular weight, charged at near-neutral pH. Therefore, the egg
gelatin is derived from the collagen present in albumin is not supposed to act as flocculant be-
animals, such as cattle, pigs, sheep, and fish. cause of similar charge with diatomite surface.
Collagen is the major insoluble fibrous protein Hence, methylated egg albumin with positive
in the extracellular matrix and in the connective charge will attach strongly to the negatively
tissue of animals (Lodish et al., 2000). Gelatin charged diatomite surface even at near-neutral
has been used in the wine-fining process for the pH and led to improved flocculation. Modern
removal of the natural haze-forming constitu- extraction techniques allow for the extraction of
ents (Cole, 1986). Similarly, Piazza and Garcia albumin from hemolyzed blood as well as from
(2010b) showed that porcine gelatin promoted high-quality plasma.

1.  General introduction

 REFERENCES 33
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