Food Hydrocolloids xxx (2017) 1e8

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Food Hydrocolloids
journalhomepage:www.elsevier.com/locate/foodhyd

Isolation, characterization and functionalities of bio-fiber gums isolated from

grain processing by-products, agricultural residues and energy crops*

Madhav P. Yadav*, Kevin B. Hicks

Sustainable Biofuels and Co-Products Research Unit, Eastern Regional Research Center, Agricultural Research Service, USDA, 600 East Mermaid Lane, Wyndmoor,
PA 19038, USA

article info abstract

Article history: Potentially valuable water soluble bio-fiber gums (hemicellulose B) were prepared from low-valued grain processing by-
Received 21 December 2016 products, agricultural residues and energy crops by an alkaline extraction followed by ethanol precipitation. The bio-fiber gum
Received in revised form 13 (BFGs) from all these sources are very rich in soluble dietary fiber (87.2e93.6%) except from wheat bran and sugarcane
March 2017 Accepted 7 April
bagasse (only 60.3 and 55.5% respectively) and they are predicted to be non-caloric in human diets qualifying as non-caloric
2017
food ingredient in many products. The sugar composition of these BFGs showed that they were typical arabinoxylans
Available online xxx
containing high molar ratio of arabinose to xylose with some other sugars such as galactose, glucose, galacturonic acid and
glucuronic acid in the side chains. The yield of pure BFG from these plant materials varied from 2.33 (rice fiber) to 25.13%
Keywords:
(corn bran) and they all, except BFG from wheat bran, are very good emulsifiers for stabilizing oil in water emulsions. Their
Soluble dietary fiber
Agricultural processing by-products ORAC values ranged from about 7000 to 29,000 m mole Trolox/ 100 g sample showing that they have the ability to provide
Emulsion stability Emulsifier antioxidant activity in foods, as well as of-fering the other functionalities and health-promoting benefits of dietary fiber. The
ORAC values of BFGs are higher than their respective original plant materials indicating that the antioxidant-rich portions of
Antioxidants the various plant materials can be solubilized and fractionated by the present alkaline extraction process. Published by Elsevier
Ltd.

1. Introduction renewable resource available in a large quantity, can be a good source of

The grain processing by-products (corn bran/fiber, wheat bran, rice fiber
etc.), agricultural residues (corn stover, wheat straw, sugarcane bagasse etc.)
and lignocellulosic energy crops (switch-grass, miscanthus etc.) are abundant
and often low-valued agri-cultural materials produced in USA and other parts
of the world. They are natural composites of three major polymeric
components: cellulose, hemicelluloses and lignin, which are linked together
via covalent and hydrogen bonds. Along with the three major poly-meric
components, namely cellulose, hemicellulose, and lignin, they may also
contain other minor components, such as extractives (phenolics, lipids, etc.),
starch, pectins and proteins (Parajo, Garrote, Cruz, & Dominguez, 2004;
Monlau et al., 2012). Corn fiber/bran, a
* cholesterol and maintain a healthy bacterial population. Diets high in fiber-
Mention of trade names or commercial products in this article is solely for the purpose of
providing specific information and does not imply recommendation or endorsement by the U.S.
Department of Agriculture.
* Corresponding author.
E-mail address: madhav.yadav@ars.usda.gov (M.P. Yadav).
valuable consumer products, as for an example corn fiber gum (Yadav,
Johnston, Hotchkiss, & Hicks, 2007a, 2016a, b). These agricultural materials
are the source of many bio-based products, which can be used in many food
and non-food industries. The fibrous materials from lignocellulosic sources
have various appli-cations, such as building materials, particle board, human
food, animal feed, cosmetics, medicines and many others (Reddy & Yang,
2005; Reddy et al., 1989). They are also a very good source of dietary fiber.
Dietary fibers are complex carbohydrates resistant to diges-tion by human
digestive enzymes. They can be classified into an insoluble group and a
soluble group. The soluble dietary fiber (SDF) helps to suppress blood
cholesterol levels, especially LDL-cholesterol, by binding to bile acids, which
are made from choles-terol, in the gastrointestinal tract and carrying them out
of the body as waste (Woo & Kim, 2005). It also inhibits lipid absorption in
the digestive tract. In addition, fermentation of SDF in the colon gen-erates
short chain fatty acids (SCFA) which can suppress the syn-thesis of
rich carbohydrates can improve glucose and

http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
0268-005X/Published by Elsevier Ltd.

Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
2 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8
insulin level and also contribute to a decrease in blood lipids for people with
Type II diabetes (Chandalia et al., 2000). However, the levels of fiber required
to induce these beneficial effects are high (up to 35 g/day) (Franz et al., 2002),
which may be difficult for people accustomed to Western diets to achieve. For
this reason, the development of SDF has gained increasing interests (Groop,
Aro, Stenman, & Groop, 1993; Vuksan et al., 1999; Wursch & Pi-Sunyer,
1997). Arabinoxylan-rich fiber (hemicellulose B) has drawn special attention
because of its unique acceptability and palatability (Lu, Walker, Muir,
Mascara, & ODea, 2000a; Lu, Gibson, Muir, Fielding, ODea, 2000b, 2004).
Arabinoxylan has a beta-1,4-linkage that cannot be hydrolyzed by human
digestive enzymes, and thus is a good dietary fiber.
In addition to above uses, lately lignocellulosic materials are also being
used for making cellulosic ethanol. Cellulosic biomass con-tains about one
third hemicelluloses A and B (Hoije, Grondahl, Tommeraas, & Gatenholm,
2005), in which Hemicellulose B (Hemi. B) contains mainly carbohydrate
biopolymer arabinoxylan with some other minor components. Arabinoxylan
has a linear 1,4 linked xylan backbone with a-L-arabinose, galactose,
glucuronic acid and some other sugars in the side chains. Hemi. B from corn
fiber has been isolated and its structure function relationship is well studied
(Doner & Hicks, 1997 and 1998 and Doner & Hicks, 1997; Yadav et al.,
2007a, b, c, d; 2008). Hemi. B from corn fiber (also called corn fiber gum,
CFG) is found to be a very good emul-sifier for the oil-in-water emulsion
system. It contains minor, but functionally significant amounts of protein,
phenolic acids and lipids, which contribute towards it emulsifying properties
and may also provide nutraceutical value (Yadav et al., 2007a, b, d and 2008).
The insoluble cellulosic residue remaining after hemicellulose sol-ubilization
of corn fiber gum has also been processed and isolated in 25% yield (Doner &
Johnston, 2001; Yadav, Kale, Hicks, & Hanah, 2017). The functional
hemicellulose B from barley hulls, several varieties of barley straws and
several sorghum materials has been isolated, characterized and studied
(Yadav & Hicks, 2015, Qiu, Yadav, & Yin, 2017). The effects of CFG on the
pasting, thermal and gelatinization behaviors of starch have also been well
studied (Qiu et al., 2015; Qiu et al., 2016).
In recent years, bioethanol production from lignocellulosic biomass is
hoped to be a very important alternative to gasoline for transportation as it can
reduce greenhouse CO2 emissions. Another big advantage using
lignocellulosic biomass for biofuel production is that it does not complete
with food supplies. The commercially available biofuel of today is still mostly
produced from edible feedstocks, such as corn starch and sugarcane juice,
which can sometimes compete for their food and feed uses. Since lignocellu-
losic materials are non-food biomass they may have less impact on food
and/or feed markets. These biomasses may have a consider-able amount of
alkali soluble, non-cellulosic carbohydrate poly-mers. Such alkali soluble
carbohydrates are usually polymers of pentoses such as xylose and arabinose
which are not easily fer-mented to biofuel in high yield as are sugars such as
glucose, found in the insoluble cellulose-rich fibers. Removal of these alkali
soluble polymers from the cellulosic biomass would yield a cellulose-rich
(and glucose-rich) feedstock that could be easily hydrolyzed and easily
fermented by well-known microorganisms such as brewers yeast,
Saccharomyces cerevisiae. Thus our current studies are focused on
developing valuable uses for these alkali soluble poly-saccharides, so they can
become valuable coproducts of the cellu-losic ethanol process, improving the
overall economics for cellulosic biofuel production. In this work, we focus on
food grade hemicellulose B, also called bio-fiber gum (BFG) from grain pro-
cessing by-products, agricultural residues and energy crops by treating them
with alkali, characterizing them and study their emulsifying and antioxidant
properties.
Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
Note: This manuscript has been written to honor and celebrate Prof. Glyn
O. Phillipss 90th Birthday for his excellence and invaluable contribution in
the field of hydrocolloids.
2. Materials and methods
2.1. Materials
The plant materials were obtained from various sources as fol-lows: Corn
bran (Bunge Milling, Inc., St. Louis, MO), Corn stover (Lawrence Shrawder,
a Private Farm, Kempton, PA), Rice fiber (SunOpta Ingredients, Cedar
Rapids, IA), Wheat bran (Deibel Lab-oratories, Bethlehem, PA), Wheat straw
(Hougar Farms LLC, Coat-esville, PA), Switchgrass (ARS lab at University
Park, PA), Miscanthus (University of Illinois at Urbana-Champaign, Cham-
paign, IL) and Sugarcane bagasse (Southern Regional Research Center, ARS,
USDA, New Orleans, LA). They were oven dried and ground to a 20-mesh
particle size using a Wiley mill. Termamyl a-amylase was a gift from
Novozymes, Davis, CA. Medium Chain Triglyceride (MCT) oil was
purchased from Nestle Nutrition, Min-neapolis, MN. Sodium hydroxide and
hydrochloric acid were ob-tained from Sigma-Aldrich (St. Louis, MO). All
chemicals were reagent grade.
2.2. Standard proximate analyses
Protein (N 6.25), ash and moisture contents of all samples were
determined using AACC Approved Methods 46-30, 08-01 and 44-19
respectively (AACC International, 1995). Crude fat content was determined
using the ANKOM XT10 Extractor with hexane as the extracting solvent.
Starch content was determined using a total starch assay kit (Megazyme, Inc.,
Wicklow, Ireland) based on the use of thermostable a-amylase and
amyloglucosidase (McCleary, Gibson, & Mugford, 1997).
2.3. Dietary fiber determination
Insoluble dietary fiber (IDF), soluble dietary fiber (SDF), total dietary fiber
(TDF) and neutral detergent fiber (NDF) in CAF were determined following
the standard A2000 automated fiber analyzer procedure (ANKOM
Technology, 2011).
2.4. Isolation of hemicelluloses A and B
The hemicellulose A (Hemi. A) and hemicellulose B (Hemi. B) were
extracted from the plant material according to the procedure of Yadav, Cooke,
Johnston, & Hicks, 2010 with some modification. In brief, the ground plant
material was boiled at 85 C with efficient mechanical stirring in the presence
of heat stable Termamyl a-amylase at pH 6.8 for 1 h to hydrolyze starch.
Then the pH of this suspension was raised to 11.5 by adding 50% NaOH and
the boiling and stirring were continued for additional 30 min. During the re-
action, pH was maintained at 11.5 by adding 50% NaOH and the reaction
volume was maintained by adding water as needed to compensate water loss
due to evaporation. The hot slurry of the deconstructed material was
immediately sheared using a high speed Polytron (PT 10/35 GT) equipped
with 12 mm probe (Kine-matica Inc., Bohemia, NY) at 10,000 rpm for 30 min
and cooled to room temperature. The solid residue was separated from the re-
action mixture by centrifugation at 14,000 g for 10 min and discarded. The
supernatant was collected in a beaker and its pH was adjusted to 4.0e4.5 by
adding concentrated HCl to precipitate acid insoluble Hemi. A, which was
collected by centrifugation at 10,000 g for 30 min. Two volumes of ethanol
were gradually added to the supernatant with stirring to precipitate the acid
soluble
 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8
Hemi. B (also called bio-fiber gum), which was collected and dried in the
vacuum oven.
2.5. Determination of sugar composition
Sugars were analyzed by HPAEC-PAD after hydrolyzing Hemi. A and B
samples into monosaccharides by methanolysis combined with TFA treatment
as explained in detail by Yadav, Johnston, & Hicks, 2007b.
2.6. Emulsion preparation and particle size determination
The sample for emulsification (2.5 g) was prepared with 1:4 ratio of
emulsifier to MCT (octanoic/decanoic acids triglycerides) containing 0.1%
sodium benzoate and 0.3% citric acid. A stock so-lution of emulsifier (27.78
mg/g solution) containing 0.1% (w/w) sodium benzoate (a preservative) and
0.3% (w/w) citric acid was prepared for emulsification study by slowly adding
the calculated amount of Hemi. B, a little at a time with vigorous stirring to a
solution of sodium benzoate and citric acid in water at room tem-perature and
then gently stirring overnight to produce a hydrated, well dissolved and
homogeneous solution. The samples for oil-in-water emulsions were prepared
in triplicate for each sample by taking 2.25 g of above emulsifier stock
solution and 0.25 g of MCT (octanoic/decanoic acids triglycerides) in a glass
vial. No weighting agent was added during emulsion preparation to avoid the
effects of such agent on the emulsification process. The solution was vor-
texed and then homogenized using a polytron bench top homog-enizer
equipped with a 12 mm diameter head (Brinkmann, Switzerland, PT 10/35) at
20,000 rpm for 3 min. The above ho-mogenized emulsion was passed through
the EmulsiFlex-B3 high-pressure homogenizer (Avestin Inc., Canada) at
20,000 psi ho-mogenization pressure 3 times to prepare the final emulsions.
The particle size distribution of the emulsions was measured using a laser
diffraction particle size analyzer (Horiba LA-950). The volume mean
diameter was used for describing particle size of emulsions. The
emulsification effectiveness was evaluated on the initial par-ticle size of the
emulsion at 0 time. The emulsion stability was determined after storing
emulsions at 60 OoC (acceleration test), (Al-Assaf, et al., 2007) for 3 and 7
days by measuring their particles size.
2.7. Antioxidant activity measurement
The antioxidant activity was measured by oxygen radical absorbance
capacity (ORAC) assay following the published pro-cedure (Huang, Ou,
Hampsch-Woodill, Flanagan, & Prior, 2002) with some modification (Ou et
al., 2002).
3. Results and discussion
3.1. Compositional analysis of the original plant materials
The compositional analysis of the original plant materials is given in
Table 1. These samples contain about 1.58e10.27% mois-ture and
0.95e11.8% ash (dry weight basis). The protein content varies from 0.63 to
18.33%. The wheat bran contains the highest percent of protein (18.33%).
Corn fiber and corn stover have 6.27 and 5.03% of protein respectively, which
are higher than all remaining materials (0.63e2.16%). Rice fiber has the least
percent of protein (0.63%) among all the plant materials studied. All of these
materials contain less than 4% (0.20e3.52%) of extractable lipids (crude fat),
but they have very high levels (80.67e90.40%) of neutral detergent fiber,
except wheat bran (45.44%), indicating the presence of complex non-starch
polysaccharides and lignin-type of
Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
3
molecules. They contain about 0.30e8.97% starch except wheat bran, which
has 21.87%, indicating that a significant amount of endosperm was not
removed during milling. Since the wheat bran is very rich in starch content, it
has the lowest percent (41.2%) of insoluble dietary fiber (ISD) in comparison
to all other plant ma-terials studied (84.0e93.1%). They all have very low
level of soluble dietary fiber (0.3e3.6%) but their total dietary fiber content is
above 82% except wheat bran (44.8%).
3.2. Isolation of Hemi. A and B
The Hemicelluloses A and B were isolated from grain processing by-
products (corn bran, wheat bran and rice fiber), agricultural residues (corn
stover, wheat straw and sugarcane bagasse) and energy crops (switchgrass
and miscanthus) by an alkali treatment following the scheme shown in Fig. 1.
The percentage yield of Hemi. A and B on dry weight basis is shown in Table
2. The starch asso-ciated with the original plant materials was hydrolyzed to
malto-dextrins by treating them with heat stable a-amylase at 85 C to avoid
the contamination of alkali soluble starch in hemicelluloses and making them
calorie free. In a biofuel process, this hydrolyzed starch could be utilized to
produce fuel ethanol.
All these plant materials contain a high percent of acid insoluble
hemicellulose, called Hemi. A (11.20e22.63%) except corn bran, which has
only 3.98% and is comparable to the previous report (Yadav et al., 2007a).
Hemi. A from these sources has not been well characterized and its functional
properties are not known. It is likely that the Hemi. A fraction contains a
variety of components, not just pure polysaccharide. A full characterization
and study of its functionality would be needed to determine applications and
value for this fraction. The corn bran has comparatively more alkali and acid
soluble Hemi. B (25.13%) than the other sources (2.33e14.60%). Hemi. B
from corn kernels has been shown to have functional properties that are useful
in food applications, such as being a good oil-in-water emulsifier. The
structure/function relationship of Hemi. B from corn fiber is well studied and
reported (Yadav et al., 2007a, b, c, d, 2008). The corn stover, rice fiber, wheat
straw, switchgrass, miscanthus and sugarcane bagasse do not contain a very
high percentage of Hemi. B (2.33e8.85%), but if functionally useful, it could
be of considerable importance, especially if it could be obtained as a valuable
co-product during the pre-treatment of these materials prior to their
saccharification and fermentation for fuel ethanol production.
3.3. Proximate composition and dietary fiber analysis of Hemi. A and B
Table 3 shows the proximate composition of Hemi. A isolated from all
eight sources. The ash content in these samples was high ranging from 8.25 to
20.05%. The Hemi.A from corn bran, corn stover and wheat bran are very
rich in protein containing 24.58, 13.51 and 37.85% respectively but the Hemi.
A isolated from the remaining five sources contain only 2.72e4.41% of
protein. Corn bran and wheat bran had 03.91 and 1.43% residual starch but
for all other samples the starch content was less than 1%. Their crude fat
content varied from 0.65 to 13.30%, except miscanthus (0% fat). The neutral
detergent fiber (NDF) content in these samples varies from 0 to 0.69%, except
one from corn stover (4.98%). As the Hemi. A from all given sources has
high level of combined amount of ash, protein and fat, they do not contain
very high percent of ISD (1.6e22.5%), SDF (11.2e35.6) and TDF
(15.6e54.0%).
Table 4 shows the proximate composition of Hemi. B isolated from the
above mentioned sources. Though they were produced by ethanol
precipitation, their ash content is still high (2.97e24.39%), showing the
precipitation of some salt with ethanol. Their protein
4 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8

Table 1
Proximate composition and dietary fiber content of original plant materials (all dry weight basis).

Sources Moistureb Ashb Proteina,b Starchb NDFb Crude Fatb ISD SDF TDF
Corn bran 7.87 0.09 0.95 0.04 6.27 0.06 8.97 0.26 80.98 0.69 3.00 0.10 81.9 0.3 82.3
Corn stover 6.24 0.03 5.43 0.18 5.03 0.11 0.67 0.05 80.67 0.47 1.28 0.52 83.5 3.6 87.2
Rice fiber 4.00 0.07 2.41 0.04 0.63 0.06 0.43 0.07 88.54 1.45 0.20 0.07 93.4 3.6 97.1
Wheat bran 10.27 0.02 5.66 0.05 18.33 0.23 21.87 0.73 45.44 0.33 3.52 0.27 41.2 3.6 44.8
Wheat straw 1.58 0.09 6.07 0.10 2.16 0.03 0.60 0.10 85.72 0.42 1.86 0.24 85.2 2.0 87.3
Switchgrass 8.99 0.07 2.38 0.11 2.08 0.01 0.65 0.11 87.06 0.03 0.86 0.17 93.1 1.2 94.3
Miscanthus 2.09 0.17 3.30 0.01 1.31 0.04 1.54 0.14 86.22 1.51 0.72 0.10 87.2 1.6 88.9
Sugarcane bagasse 2.35 0.08 11.80 0.65 1.64 0.06 0.30 0.05 90.40 0.75 1.00 0.19 84.0 1.4 85.4
a Calculated by multiplying nitrogen content by 6.25.
b
Average of triplicates.

Ground Plant Material

Suspended in 85 OC water, pH adjusted to 6.8 and stirred with -
amylase for 1 hr. Then pH raised to 11.5 and stirred at for 30 min.
Slurry of Deconstructed Material

High speed shearing for 1 hr and centrifugation

Supernatant (Alkaline extract) Solid Residue (alkali extracted), Either Discarded or

Used to Cellulosic Arabinoxylan(CAF) Preparation
pH 4.0-4.5

Supernatant (alkali extract)

Hemi. A

2X EtOH

Precipitate-Hemi. B (Bio-Fiber Gum)

Fig. 1. Basic scheme for the isolation of hemicelluloses from corn bran, wheat bran, corn stover, rice fiber, wheat straw, switchgrass, miscanthus and sugarcane bagasse.

Table 2 diets, and should be very useful for making non-caloric food
Hemicellulose A and B from plant materials (% yield, dry weight basis). products.
Sources Hemi. A Hemi. B

Corn bran 3.98 25.13

3.4. Sugar composition of Hemi. A and B
Corn stover 19.82 8.85
Rice fiber 11.20 2.33
Wheat bran 14.85 14.60 Table 5 shows the sugar composition (relative mole percent) of
Wheat straw 22.63 4.80 the Hemi. A isolated from all eight plant materials. The acid insol-
Switchgrass 18.35 5.78 uble Hemi. A from all these sources look like arabinoxylan with
Miscanthus 17.03 2.56
varying Ara to Xyl ratio of 0.0.19 to 0.98 and a few percent of Rha,
Sugarcane bagasse 17.08 3.03
Gal, Glc and acidic sugars. Though there is some similarity in its
sugar composition with Hemi. B, it differs greatly in its solubility
under acidic conditions. Ara to Xyl ratio in Hemi. A from corn bran
content varied from 0 to 1.27% except Hemi. B from wheat bran,
and wheat bran is 0.73 and 0.98 respectively, showing highly
which was rich in protein (8.37%). They all had a little bit of residual
branched structure of their arabinoxylans. But the Ara to Xyl ratio in
starch (0.54e2.43%) and crude fat (0.14e1.37. As expected, these
the Hemi. A from other sources varies from 0.19 to 0.32 showing a
samples had 0 or very low percentage of NDF (0.00e1.92%) and ISD
less branched arabinoxylan structure.
(0.00e3.2%). But they were very rich in SDF varying from 87.2 to
Table 6 shows the sugar composition (relative mole percent) of
93.6% except Hemi. B from wheat bran and sugarcane bagasse,
the Hemi. B isolated from the same eight sources. Hemi. B isolated
which had only 60.3 and 55.5% SDF respectively. It is clear that
from all these sources show a typical arabinoxylan structure with
Hemi. B isolated from most of these sources were almost pure
xylan backbone and Ara in the side chains. The Ara to Xyl ratio of
soluble dietary fiber. They are predicted to be non-caloric in human
Hemi. B from corn bran and wheat bran are high (0.57 and 0.68

Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8 5

Table 3
Proximate composition and dietary fiber content of Hemicellulose A isolated from plant materials (all dry weight basis).

Sources Moistureb Ashb Proteina,b Starchb NDFb Crude Fatb ISD SDF TDF
Corn bran 1.38 0.16 9.22 0.27 24.58 0.67 3.91 0.48 0.00 10.98 0.82 8.2 19.1 27.3
Corn stover 7.95 0.14 9.60 0.17 13.51 0.48 0.42 0.18 4.98 0.55 3.47 0.37 13.6 26.0 39.4
Rice fiber 2.38 0.17 18.36 0.36 4.30 0.11 0.37 0.15 0.34 0.93 0.68 0.74 19.2 34.8 54.0
Wheat bran 2.66 0.10 8.15 0.18 37.85 0.67 1.43 0.16 0.69 0.19 13.30 0.48 4.3 11.2 15.6
Wheat straw 2.43 0.05 14.35 0.90 4.37 0.35 0.45 0.21 0.00 0 0.79 0.41 2.8 35.6 38.3
Switchgrass 6.39 0.13 13.36 0.47 4.41 0.17 0.60 0.08 0.24 0.08 1.80 0.15 1.6 35.4 37.0
Miscanthus 0.92 0.01 12.42 0.65 2.72 0.06 0.99 0.06 0.00 0.00 9.5 30.4 39.9
Sugarcane bagasse 2.03 0.18 20.05 0.56 3.84 0.02 0.40 0.08 0.00 0.65 0.40 22.5 22.4 44.8
a Calculated by multiplying nitrogen content by 6.25.
b
Average of triplicates.

Table 4
Proximate composition of pure bio-based fiber gums (Hemicellulose B) isolated from plant materials (all dry weight basis).

Sources Moistureb Ashb Proteina,b Starchb NDFb Crude Fatb ISD SDF TDF
Corn bran 3.44 0.21 2.97 0.04 1.20 0.04 1.69 0.14 0.00 0.72 0.12 1.4 93.6 95.1
Corn stover 5.34 0.14 6.92 0.02 1.27 0.14 1.40 0.05 0.26 0.01 0.42 0.17 0.0 87.2 87.3
Rice fiber 7.21 0.32 7.69 0.22 0.43 0.07 0.54 0.11 0.00 ND 1.6 90.3 91.9
Wheat bran 4.21 0.12 4.39 0.09 8.37 0.29 2.36 0.07 1.92 0.52 0.14 0.13 0.6 60.3 61.0
Wheat straw 5.36 0.08 6.21 0.15 0.00 0.59 0.12 0.00 0.80 0.31 0.60 93.1 93.7
Switchgrass 5.02 0.13 5.16 0.02 1.14 0.08 0.90 0.04 0.43 0.29 1.01 0.48 0.7 93.0 93.7
Miscanthus 4.08 0.33 7.10 0.04 0.00 2.43 0.11 0.00 1.37 0.45 3.2 88.0 91.1
Sugarcane bagasse 3.64 0.30 5.98 0.06 0.00 1.98 0.04 0.00 0.44 0.65 1.0 55.5 56.7
a Calculated by multiplying nitrogen content by 6.25.
b
Average of triplicates.

Table 5 3.5. Emulsification study of Hemi. B

Carbohydrate composition of Hemicellulose A isolated from plant materials (relative mole %).

The emulsion stability study was performed at 2.5 g scale and the particles
Sources Rha Ara Gal Glc Xyl GalA GlcA Total (droplets) size (diameter) was measured using a laser diffraction particle size
Corn bran 0.00 35.26 5.29 3.25 48.62 0.0 7.48 100.00 analyzer. Medium chain triglycerides (MCT) (density, 0.95 g/ml at 20 C)
Corn stover 1.65 16.55 7.67 16.71 52.16 1.85 3.40 100.00 were used as the dispersed phase since it is stable to oxidation with no bad
Rice fiber 0.28 14.62 2.40 4.14 77.72 0.03 0.81 100.00 smell or aroma and dissolves many lipophilic substances. The average particle
Wheat bran 0.07 27.32 4.05 36.67 27.86 0.00 4.03 100.00 size of emulsions prepared with Hemi. B from all eight plant materials are
Wheat straw 0.58 14.66 2.76 5.58 74.57 0.74 1.10 100.00
shown in Fig. 2. The emulsification effectiveness was evaluated by measuring
Switchgrass 1.31 15.35 2.69 6.18 73.35 0.00 1.11 100.00
Miscanthus 0.35 14.56 1.67 7.18 75.24 0.21 0.79 100.00 the initial particle size immediately after making emulsions (0 day). The
Sugarcane bagasse 0.41 13.97 2.48 7.23 75.01 0.00 0.91 100.00 emulsion stability was evaluated by storing the emulsions at 60 C (accelerated
Note: Rha, Rhamnose; Ara, Arabinose; Gal, Galactose; Glc, Glucose; Xyl, Xylose; GalA,
temperature stress test) and measuring the particle size after 3 and 7 days. As
Galacuronic acid; GlcA, Glucuronic acid. shown in Fig. 2A, Hemi. B from all sources makes very effective initial
emulsions on 0 day with average particle size of 1 or below 1 mm, except
from wheat bran, whose average particle size was 2.22 mm. The average
particle size of emulsions prepared using Hemi. B from corn stover, rice fiber,
respectively) like their corresponding Hemi.A, showing their highly branched
wheat straw, switchgrass, miscanthus and sugarcane bagasse rose only to
structures. The Hemi. B from other sources have lower Ara to Xyl ratio
showing less branched structures. The Ara to Xyl ratio (branching of AX) of about 1 or a little bit above 1 mm after 3 days storage at accelerated
Hemi. B from corn bran agree with the reported results (Yadav et al., 2007a). temperature conditions. But the average particle size of emulsions prepared
with Hemi. B from corn bran and wheat bran
The other sugars present in the Hemi. B from all these sources are Rha, Gal,
Glc, GalA and GlcA.

Table 6
Carbohydrate composition of Bio-based Fiber Gums (Hemicellulose B) isolated from plant materials (relative mole %).

Sources Fuc Rha Ara Gal Glc Xyl GalA GlcA Total

Corn bran 0.19 0.43 27.41 12.06 4.27 48.43 1.02 6.20 100.00
Corn stover 0.84 1.32 17.99 10.85 9.58 52.25 1.75 5.42 100.00
Rice fiber 0.20 0.97 18.12 8.06 3.64 65.36 1.57 2.07 100.00
Wheat bran 0.00 0.55 31.10 5.27 14.00 46.03 0.83 2.21 100.00
Wheat straw 0.28 2.50 19.67 11.67 7.25 54.46 1.69 2.50 100.00
Switchgrass 0.33 1.12 19.04 7.25 6.33 61.69 1.67 2.57 100.00
Miscanthus 0.31 1.00 18.46 7.99 10.17 57.89 1.90 2.28 100.00
Sugarcane bagasse 0.19 1.28 17.12 6.37 12.38 58.39 2.16 2.11 100.00

Note: Fuc, Fucose; Rha, Rhamnose; Ara, Arabinose; Gal, Galactose; Glc, Glucose; Xyl, Xylose; GalA, Galacturonic acid; GlcA, Glucuronic acid.

Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
6 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8

Fig. 2. Average particle size of emulsions prepared with biofiber gum from corn bran, corn stover, rice fiber, wheat bran, wheat straw, switchgrass, miscanthus and sugarcane bagasse. Lower particle

size is an indication of greater emulsion stability. Each plotted point is an average of three trials SD. A 0 day, B 3 days and C 7 days.

rose to 2.80 and 14.88 respectively during 3 days storage at the accelerated (Fig. 2B). After 7 days storage at the same temperature, the average particle
temperature (60 C) showing that they made a less stable emulsions in size of Hemi. B from wheat bran increased drastically above 40 mm, showing
comparison to Hemi. B from other six sources very unstable emulsions, which could be due to

Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8
35,000
30,000
Whole Material, Hemicellulose-B
25,000
20,000
15,000
10,000
5,000
Fig. 3. ORAC values of the whole biomass and the Hemicellulose-B (BFG) isolated from them.
very high content of ash (24.39) in this sample. The particle size of the
emulsion prepared with Hemi. B from corn bran was about 9 mm, showing
an intermediate stability as reported previously (Yadav et al., 2007a). But the
particle size of emulsion prepared with Hemi. B from all remaining six
sources stayed below 3 mm, indicating that they were very good emulsifiers
and make stable emulsions. The emulsion stability of these samples did not
correlate well with their protein content showing that their emulsifying
properties were not just based on the protein content but some other structural
features were responsible for their emulsifying properties.
3.6. Antioxidant properties of BFG and its source material
By normal cellular processes in the body, UV irradiation and
environmental stresses, highly reactive molecules like free radicals and
reactive oxygen species are generated. These reactive mole-cules react with
cellular components damaging DNA, carbohy-drates, proteins and lipids
causing injury to cells and tissues. Excess production of such reactive species
can cause several diseases including cancer, diabetes and atherosclerosis etc.
Most organisms have antioxidant systems to protect themselves from
oxidative stress, however an excess free radical and/or reactive oxygen spe-
cies can cause severe damage. One way to measure the antioxidant power of
compositions, foods, and plant phytochemicals is to determine the Oxygen
Radical Absorbance Capacity (ORAC) value of the composition (Graf, 1992;
Ohta, Yamasaki, Egashira, & Sanada, 1994). The ORAC antioxidant assay
measures the loss of fluorescein fluorescence over time due to peroxyl-radical
formation by the breakdown of 2,2-azobis-2-methyl-propanimidamide,
dihydro-chloride (AAPH). Trolox, which is a water soluble vitamin E analog,
serves as a positive control inhibiting fluorescein decay in a dose dependent
manner. The ORAC assay is a kinetic assay measuring fluorescent decay and
antioxidant protection over time. The anti-oxidant activity can be normalized
to equivalent Trolox units to quantify the composite antioxidant activity
present. This assay measures the materials antioxidant activity by hydrogen
atom
Please cite this article in press as: Yadav, M. P., & Hicks, K. B., Isolation, characterization and functionalities of bio-fiber gums isolated from grain processing
by-products, agricultural residues and energy crops, Food Hydrocolloids (2017), http://dx.doi.org/10.1016/j.foodhyd.2017.04.009
7
transfer. The higher the ORAC score is an indication of a greater the
materials antioxidant capacity. There is great need of compounds that can
serve to protect foods and non-food products against oxidative damage due to
free radicals, oxygen, ozone, and other deleterious environmental agents. It
has been reported that the free and bound phenolic residues associated with
the plant poly-saccharides, like spruce galactoglucomannans,
carboxymethylated spruce galactoglucomannans, gum arabic and corn fiber
gum effi-ciently inhibited and altered the pathways of lipid oxidation of
rapeseed oil in its water emulsion in the presence of each of the above
polysaccharides (Lehtonen et al., 2016). BFGs, which have ability to
emulsify, encapsulate, or form a protective film on the products that are
sensitive to oxidation, such as vitamins, omega-3 polyunsaturated fatty acids,
fish oils, medicines, paint pigments, and the like, should be useful for
protecting these sensitive mate-rials (Yadav et al., 2016a, b). BFGs isolated
from these eight plant sources do not only have the ability to emulsify,
encapsulate, and form films on these sensitive oxidation-prone materials, but
also have the ability to prevent oxidation due to their antioxidant properties.
The hydrophilic ORAC values of pure BFGs (Hemi. B) isolated from corn
bran, wheat bran, corn stover, rice fiber, wheat straw, switchgrass,
miscanthus, and sugarcane bagasse are higher than their respective original
biomasses (Fig. 3) indicating that more antioxidant containing BFG products
can be solubilized and made available by the present alkaline extraction
process. Our re-sults clearly indicate that all these BFGs, especially those
from wheat bran, corn stover, switchgrass and miscanthus, are very rich in
antioxidant (ORAC Value, above 7000 mmol TE/100 g), which is higher than
the original materials they were prepared from. In addition to being very
powerful antioxidants, these BFGs are very rich (over 80%) in soluble dietary
fiber (Table 4). Human con-sumption of such materials could be beneficial for
prevention of heart disease.
4. Conclusion
Bio-fiber gum (BFG), also called Hemicellulose B is a highly
8 M.P. Yadav, K.B. Hicks / Food Hydrocolloids xxx (2017) 1e8
functional value-added product that can be prepared with the yield varying
from 2.33 to 25.13% from many low value grain processing by-products,
agricultural residues and energy crops. The BFG con-sisted almost entirely of
soluble dietary fiber, making them suitable as non-caloric bulking and
texturizing agents in many food prod-ucts providing them with potential
health benefits like blood cholesterol level lowering and prebiotic activities in
the human gut. All these materials contained a high percentage of acid
insoluble Hemi. A (3.98e22.63%), which needs more study to determine
functionality and potential new industrial applications. Corn and wheat bran
contained comparatively more Hemi. B (25.13 and 14.60% respectively) than
other sources (2.33e8.85%) and Hemi. B from all these sources has a typical
cereal arabinoxylan structure with xylan backbone and Ara in the side chains.
A low percent of Gal, Glc and acidic sugars was also detected and more likely
they are present in the side chains. They have highly branched structure.
Hemi. B from all these sources are very good emulsifiers except the Hemi. B
from wheat bran and they are very good antioxidant with high ORAC. Owing
to their high functionality and non-caloric na-ture, BFGs produced from these
low-valued agricultural by-products are very interesting and should be
commercially valu-able health-promoting, soluble dietary fiber based bulking
agents, viscosifiers and texturizing agents for use in the food industry and
beyond.
Acknowledgements
We would like to acknowledge Stefanie Simon for her excellent technical
assistance in preparing these BFGs and charactering and doing their
functionalities test.
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