Processing of Brewer’s Spent Grain

Technology Review

BSE 4125: Senior Design

November 5, 2017

Team BSG
Lucy Epshteyn, Samuel Villarreal, Zhicheng Xu, Ryan Mitchell

Faculty Advisor: Haibo Huang

Cargill Industry Advisor: Sun Min Kim
Introduction
The world currently contains 7.6 billion people, with an expectancy to rise to 9.8 billion by
2050 (“World Population”, 2017, para. 1). An increase of over two-billion people within the next
thirty years may seem ordinary, but with the amount of poverty and resource scarcity currently
existing in the present population, one can only imagine the resource scarcity- including
limitations to food, water, and energy that will be faced in the near future. As humans dive
further and further into studies of health, nutrition, and the effects of various diets on the body,
the need for viable proteins to sustain human and animal diets ever increases with population.
One of the most important building blocks to sustain our bodies include proteins, derived
from variations of the 20 essential amino acids. Depending on the species of animal, amino acids
can either be produced by the animal system, or obtained through digestions of plant or animal
matter. Protein is one of the most significant building blocks required to sustain human life
because humans require proteins to obtain amino acids, and amino acids are required to rebuild
the body.
The BSG team is focusing on deriving proteins from Brewer’s Spent Grains (BSG), the
leftover kernels and mash that is a waste stream of the beer processing industry. Barley is the
main feedstock used in the brewing industry and the starch is hydrolyzed and extracted for the
fermentation process. The leftover grains, accounting for up to 85% of the total by-products from
brewing are known as BSG (Witkiewicz, 2012). BSG is currently a large nuisance to the brewing
industry because the material spoils quickly and costs money to send to landfills. BSG has
approximately 70% to 80% moisture content, presenting two main issues: the high cost of
transportation and the growth of microbes. Water adds weight, and additional weight is costly to
transport. The rich polysaccharide, protein, and moisture content make BSG susceptible to
microbial growth and spoilage following production (Lynch, 2016).
Townsley (1979) found that on average spent grain constitutes 31% of the original malt
weight and 20 kg of spent grain are capable of producing about 100 liters of beer. High
production and availability of BSG result in low market prices despite some breweries producing
less BSG than others. BSG is frequently fed to cattle and other ruminants that are capable of
digestion. BSG can also be composted to return valuable nutrients to the soil, or they can be
disposed of at landfills and waste processing facilities. These solutions produce low value
products or can cost breweries additional capital.
BSG is an attractive waste source for further processing due to the material’s chemical
composition. The main components of BSG include fiber and protein, together constituting about
80% of BSG on a dry-weight basis. Non-starch polysaccharides (including fiber) constitute about
30-50% of BSG, proteins about 19-30%, lignin about 12-28%, lipids about 10%, and ash about
2-5% (Lynch, 2016). Chemical composition varies from brewery to brewery although general
proportions of protein, fiber, and fats are very similar. Numerous value added products can also
be extracted from BSG such as ferulic and p-coumaric acids, xylose, arabinose, or as raw
material for xylitol and arabitol production (Mussatto et al., 2006). There has been a range of
research done in the past on the potential value of BSG when certain components were extracted,
or even just when BSG was dried, to be used in food or biofuels. Unprocessed BSG can be oven
dried and used to bake various items (Kissell, 1979). Alternative research has been done to
develop a system to extract the phenolic compounds found in plants, shown to exhibit anti-
carcinogenic, anti-inflammatory and antioxidant activities that would benefit humans (McCarthy,
2012). The goal of extracting phenolic compounds, proteins, or both compounds simultaneously
is to incorporate into human food as healthy food ingredients and alternatives to non-nutritious
foods.
The main goal of our design is to create a high-quality feed for farm raised fish, pets, or other
animals. The team’s goal is to purify BSG to a protein purity of about 20-50%, based on the
digestion requirements of targeted animal markets. The high concentration of fiber that resides in
BSG after brewing is indigestible by fish, dogs, and animals other than ruminants. BSG can be
incorporated into foods in small amounts, however we desire to produce a product with high
protein composition and low fiber content that will cover the health and diet needs of targeted
animal groups to be determined. Utilizing BSG as a food source for animals helps to remove a
waste stream from the brewing industry while at the same time providing a nutritious, healthy,
and valuable diet for animals.
In addition to creating a high value protein product, we also aim to use the byproduct of the
protein separation process to create the biofuel wood pellets. The energy value of the fibrous
waste from processing BSG will be determined through experimentation on Virginia Tech
campus. Processing BSG using frame and filter presses will result in a wasted cake product that
is very fibrous in nature. The team will further process this fibrous waste to create wood pellets
for heating and energy use.
Processing BSG will require that the spent grain be broken down and separated into the
various components comprising spent grain. These separated components will then be filtered
and purified to reach a high protein composition. Processing operations, chemicals used, and
temperatures reached during processing can affect taste and final product protein composition
and structure. Furthermore, high purification and high protein extraction can be very costly as
unit operations become very specialized, and enzymatic reactions and microfiltration with
membrane filters are necessary. The team will design the processing of BSG constraints:
transportation to facility, processing and manufacturing costs, processing times. Our technology
review will target the range of possible solutions for our design and the team will focus on the
use of specific unit operations to target out goal. First, the team will address various methods of
transportation and compare the different possible unit operations for a processes utilizing either
enzymes or alcohol.
Transportation
Transportation is the first concern in the food industry, as transportation is the very first
“guard” to maintain high-level food quality. According to the Center for Disease Control and
Prevention, more than 48 million people incur diseases caused by foodborne illnesses, and about
0.2% to 0.3% people must be hospitalized (Iversen, 2015). Vehicle cleanliness and temperature
are the highest priority regarding transportation because the level of contamination and bacterial
growth is directly related to the sanitation of BSG. The truck used for the transportation of BSG
must meet and exceed the standard of The FDA Food Safety Modernization Act by providing
preventive controls across the food supply (FSMA). To extend the time for processing, it is
efficient to use the combination of certain salts; research from Glass (2007) states that a mixture
of sorbate, benzoate, and propionate can prevent bacterial growth. These chemicals pass
toxicology testing, meaning that use of these combined chemicals does not present safety issues.
The addition of the chemicals can also help to reduce strains on immediate transportation
requirements. By extending the time before BSG spoilage occurs, transportation and processing
can be scheduled to optimize protein and biofuel production.
Enzyme Process:
Homogenizer vs. Wet Grinding
Before BSG can be placed in bioreactors for enzymatic extraction of protein, the BSG needs
to be reduced in size and drained to decrease water content (Sežun et al., 2011). By breaking
apart the BSG, protein will be more easily hydrolyzed by the enzyme and reaction time inside
the bioreactor will be reduced.
Comparing various homogenizers on the market, high-pressure homogenizers are favored due
to easy scale-up and effective fractioning (Spence et al., 2011). A high-pressure homogenizer
would be used in the BSG process design because the homogenizer has the option to change the
final particle size. The feature creates flexibility to create a particle size that will optimize
enzymatic breakdown of BSG. Furthermore, the high pressure from the homogenizer will also
function as a cleansing step for BSG. A commonly used apparatus of homogenization that can be
employed in our purpose is the Ultra-turrax.
Typical problems with high-pressure homogenizers include a high number of passes required
to fully fraction all the material as well as constant clogging due to larger fiber size (Oksman et
al., 2014) High-pressure homogenizers are also very expensive and the equipment is heavy,
resulting in increased capital costs. An alternative option is wet grinding, a relatively new
technology that is able to uniformly grind material down to the nanometer range. The overall
advantage of wet grinding is that the BSG could be directly placed in the grinder, avoiding a
drying step. Drying of BSG via oven or superheated steam is an energy intensive operation, so
avoiding drying would reduce energy requirements and improve economic efficiency of
processing. Furthermore, heat created by friction in the wet grinding operation is absorbed by the
water, acting as an extra safety buffer for the protein (protein denatures at high temperatures). A
commonly used apparatus for wet grinding is the 4E Electric Grinders by QCG Systems, LLC.
This apparatus is able to grind BSG to 100 µm (Quaker).
Bioreactor vs. Fermenter
The complexity of this part of the procedure and the fine parameters that have to be reached
for optimal enzyme productivity call for an instrument that has wider operation parameters than a
fermenter. A bioreactor is commonly used for growth and maintenance of a cell culture and has
the capacity to run under specific conditions that can be programed into the system. A bioreactor
is capable of self-adjustment, sampling, and extraction of a system. These features are all
necessary for protein hydrolysis with BSG. Furthermore, the utilization of a bioreactor would be
more convenient as the final product will be incorporated into animal feed and there are fewer
standards and tests when no chemicals are used to extract protein. Protein extraction using
enzymes provides extra protection and insurance that no side chemical reactions will occur and
produce side products or modify the protein to an unwanted state. According to Treimo et al.
(2008), Alcalase is the most effective peptidase for solubilization of BSG proteins, with an
ability to release up to 77% of the total protein. The peptides produced by Alcalase have lower
average molecular weight than peptides produced by less effective enzymes. Processes that
combined peptidase treatment with carbohydrate degrading enzyme preparations such as
Depol740 increased the solubilization of dry matter from 30% to 43% under optimal conditions.
The results contained minor differences in protein recovery, indicating that the most cost
effective method would be the utilization of Alcalase alone to solubilize and extract the proteins
Treimo et al., 2008). Sigma-Aldrich corporation sells ALCALASE® Enzyme, Bacillus
licheniformis 500 ml for 74 dollars (Alcalase, 2017).
Producing optimal conditions for Alcalase and reducing microorganism growth is the main
function of the bioreactor. To optimize conditions for Alcalase, the pH of the system must be
adjusted to 8 (utilizing phosphate buffer) and the temperature raised to 60ºC for at least 4 hours.
If the recommended dose (1.2 µL/ gram of dry BSG) of Alcalase is used, 1000 kg of protein can
be processed per vase of Alcalase bought (Treimo et al., 2008). The capacity and model of the
bioreactor chosen will depend on the economic value of the projected protein production.
Microfiltration
Filtration is the main unit operation required to separate protein from the BSG. The filtration
step removes undesired elements including impurities and indigestible fibers. The two different
types of filtration include dead-end filtration and cross-flow filtration. Dead-end filtration is
when the solution or feed flows perpendicular to the membrane filter. Alternatively, cross-flow
filtration occurs when the feed is flowing parallel to the membrane. The most common type of
dead-end filter is the plate and frame filter press, containing multiple plates and frames lined up
in parallel. Along with the plates and frames, there is a high-pressure pump used to deliver
suspended solids onto each plate and frame filter. When the slurry flows through the frame, the
suspended solids accumulate in each individual frame (Walter, 1970). The disadvantage of this
filtration technique is that the suspended cake needs to be collected after each round of filtration
and filters require cleaning or replacement. This replacement or cleaning step takes time and
introduces potential contamination to the cake from exposure to air.
Another type of dead-end filter is called a rotary vacuum filter. It contains a large rotating
drum with a cloth. When the drum rotates, the slurry is attached to the cloth and the resulting
suspended mixture dries in the drying zone. The rotary vacuum filter system operates as a
continuous process (Doncer, 1975). Utilizing continuous processes is advantageous when
compared to the plate and frame filter, albeit the rotary vacuum filter system consumes more
energy than the plate and frame filter.
In cross-flow filtration, the feed parallel to the filter membrane passes the system. Particles
smaller than the membrane filter pore pass through filter membrane allowing the suspended
material to remain in the stream (Marinaccio, 1989). The cross-flow filtration system is
continuous and therefore has a high feed velocity. High pressure and temperature are not
required for cross-flow filtration making it a more energy efficient process compared with dead-
end filtering. Due to increased energy efficiency, more companies are starting to use cross-flow
filtration in the separation and wastewater treatment industries (Downing 1988).
For this project, microfiltration and ultra-filtration are used with the filtration system.
Microfiltration can remove the fiber, which is mostly cellulose molecules. This is because the
membrane of micro-filtration is sized from 0.1 to 10 µm, which is the size of the fiber.
Ultrafiltration is used with a membrane pore size from 0.001 to 0.1 µm, which can separate
protein particles from water and sugar (Micro, 2017). Good Laboratory Practices (GLP) is the
regulation for filtration techniques in lab scales; during the process, the team must obey these
regulations.
Centrifugation
Downstream processes often have to separate lysed cell culture media from the substance of
interest. The simplest method is differential centrifugation, which consists of applying
centrifugal force to separate substances based on their densities. Centrifugation will ultimately
create a density gradient where the heavier material will collect at the bottom. In addition, the
peptides produced by Alcalase (enzyme that will be utilized) have lower average molecular
weight than peptides produced by other less effective enzymes (Treimo et al., 2008).
Understanding that peptides produced by Alcalase have smaller molecular weights, the team
predicts that most of the protein extracted from BSG will not migrate to the bottom, but remain
in the supernatant. The recommended conditions to run the centrifuge are between 2000-6000
rotations per minute (RPM) for 10-30 minutes to separate the substance (Acton, 2013).
Centrifuges are produced for industrial applications but are not very cost effective. Kyte
Manufacturing produces industrial centrifuges that we may utilize (Our, 2017).
Supernatant Capture
Capturing the supernatant will be necessary after centrifugation in order to concentrate the
protein to a lower volume. According to J. Treimo (2008), the peptides produced by Alcalase had
lower average molecular weights than peptides produced by the less effective enzymes
mentioned in the article. It can be assumed that the vast majority of the proteins will still be in
the supernatant and not collected at the bottom of the vial after centrifugation. Supernatant will
therefore be transferred to the next unit operation (ultrafiltration) to further concentrate the
protein.
Ultrafiltration
After the proteins are extracted from BSG they need to be concentrated to increase the
proteins market value. Ultrafiltration is commonly used in bioprocesses to concentrate the
medium by restricting material over 0.1µm from passing through membrane filters. Tang (2009)
conducted a study where he found that 92% of the protein extracted from BSG was retained by
the membranes with both filter sizes of 5 and 30 kDa, however the 5 kDa retained more protein.
Ultrafiltration is commonly used in the whey protein industry, where with a single pass
concentration can be increased 20-30 times the initial concentration (Kuo, 1983). A system with
a prose membrane of less than 1 µm would be sufficient for this process; the 3 Stage Kwik-
Change Ultrafiltration System would suffice (Water, 2017).
Spray Dryer
The final protein product increases substantially in economic value if subjected to a drying
process. Drying leads to a decrease in transportation costs and feed processors can readily apply
the protein supplement to products. Spray drying is the process of converting a liquid substance
to powder by rapidly drying with hot gas (Mishra, 2016). Spray drying is commonly used in the
pharmaceutical industry and large industrial units are available for processing BSG. Mathias
(2015) published a study where spray dryers were used in BSG and noticed that readily available
proteins increased because portions of the BSG were lysed in the process. The spray dryer
process would be incorporated after proteins are concentrated in the centrifugation process (S,
1965). The spray dryer offered by VetterTec is acceptable for drying the protein extracted from
BSG (VetterTec).
Alcohol Process:
Drying Process
BSG is known to have a high moisture content (upwards of 80%) causing problems with mold
growth, fermentation, and spoilage within a short amount of time (one week or less) (Perry,
2017). BSG must be dried until a moisture content of 10% or less is reached in order to prevent
spoilage and growth of microorganisms in the material. Current processing facilities are often
times built at breweries to remove the water from brewer’s spent grains and reduce moisture
content. Presses can be used to remove water content, generally being capable of removing up to
20% of grain moisture content. Drying is required after pressing to reduce moisture content to
levels below 10% (Santos et al., 2003).
Additional drying techniques range from oven drying to the use of superheated steam. Oven
drying can be done in a variety of ways, albeit large scale processing generally utilizes conveyor
belt equipment. Oven drying or freeze drying can reduce water content in grains without
changing the composition of grains (Bartolome et al., 2002). Freeze drying is not viable for a
large scale operations due to high energy costs, and oven drying, although one of the best
methods for drying, has been known to burn the grains towards the end of the drying process
where high temperatures are reached (Lynch, 2016). Rotary drum driers used in conjunction with
superheated steam are very viable and as long as temperatures are controlled, burning is unlikely
due to efficient heat exchange technology (Ström). A final option for removing moisture content
is through membrane separation technology; BSG is mixed with water and pressurized through a
filter and vacuum, and further dried to reach moisture contents of 20% to 30% (El-Shafey et al.,
2004).
Drying techniques are necessary when utilizing alcohol for protein separation processes.
Brewer’s spent grain with low moisture content allows alcohol to be in more concentrated forms
when reacting and breaking down spent grains. Drying also allows for storage of spent grains if
they are not able to be processed immediately upon availability, or if a plant can only access the
grains once a week and is located far from the brewery. Another option for extending the life of
brewers grains with a high moisture content without removing water is to either add alcohol or
add chemicals to the grains. Chemical additives such as potassium sorbate, benzoic acid, acetic
acid, formic acid, and lactic acid can be used to maintain nutritional and overall quality of BSG
(Al-Hadithi et al., 1985).
Grinding
Mechanical grinding would be most appropriate for alcohol solubilization of BSG. Investing
in a mechanical grinding machine can increase particle surface area and the rate of solubilization
of BSG in alcohol. A machine that fits these requirements is a grain grinder; a grain grinder is
able to deliver grinded grain up to 4 mm and is highly efficient. The Bravo Feed Grinder (can be
bought on Amazon) is an excellent example.
Storage Vessel
After the water content is removed from the BSG, the grain will be grinded and solubilized in
alcohol where the carbohydrate core of the grain will begin to degrade. The grinded BSG and the
alcohol will be held in a storage vessel that will enable complete submersion of the particles and
allow for carbohydrate degradation. Acid hydrolysis will occur and is a very effective procedure
for the degradation of polysaccharides by breaking the carbohydrate bonds of the grain
(Macheiner et al., 2003). Agitation will be employed that does not affect the total nitrogen that is
solubilized. The mixture of BSG and acid should be incubated at 25°C for 48 hours to remove
77% of the protein (Crowe, 1985). Vessels with the mixing capacity that this unit operation
requires can be found at Sprinkman Corporation (Processing, 2017).
Microfiltration will occur and is of the same nature as described previously in enzyme process
unit operations.
Final drying (same process as initial drying) will occur where both biofuel byproduct and
final protein product will be dried to moisture contents of 10% or less.
Salting Out proteins
Proteins can be condensed into bigger molecules by adding salt to the solution. This process
is known as salting. Proteins are salted out as co-precipitate by ammonium sulfate because the
saturation concentration provides high molarity that causes precipitation of most proteins
(Nehete et al., 2013). The salted solutions are also bacteriostatic and protect most proteins from
denaturation in solid state (Nehete et al., 2013). The proteins in our process need to be protected
from denaturation in their solid state because, ideally, they will be spray dried at the end of the
process. However, we are not very concerned about denaturation; we just want the proteins to
maintain the minimal integrity so that the body recognizes them when consumed. The primary
shortcoming of using salting is that contaminants often precipitate with the protein of interest,
and it would add the need of more unit operations, such as centrifugation and more desalting of
proteins, to further aggregate all proteins (Duong-Ly et al., 2014).
The group considered precipitation and salting out proteins from the solution and concluded that
due to the lag time needed to process the protein within this steps that ultrafiltration would be the
best unit operation.
Byproducts and Recycle Stream
The team will conduct experiments using an enzyme and alcohol in two separate processes to
break apart BSG. Microfiltration will allow the team to pass a fluid through a membrane filter
and collect a protein solution, and a cake or residue will form. This cake is the left behind fiber
that is desired to be removed from the protein solution. The fibrous byproduct of the
microfiltration process is retained in the filter frame press and will be used to create a biofuel
product. The team aims to conduct an energy density evaluation of the fibrous byproduct through
further experimentation to assess a wood pellet market value.
Alcohol will be an additional byproduct of the solubilization of BSG and will be recycled
using a distillation tower. The team desires to recycle as much alcohol as possible to decrease the
need to continuously add alcohol for each batch of processed BSG.
When using enzymes to solubilize BSG, water will be recycled in order to reduce water
consumption. The team is currently determining how often water will be discharged to a
wastewater treatment plant, or if water filtration and cleaning will be implemented in the process
design.
Standards
Different sectors of the FDA that regulate food production are the AFSS (formed in 2003),
FDAAA (formed in 2007), and the FSMA (Administration, 2016). The AFSS deals with the
manufacturing, labeling, storage, distribution, and use of animal food. The FDA must determine
an establishment’s or product’s degree of compliance with applicable regulations. Compliance is
primarily done by state agencies using federal or state authority. The transportation segment of
the feed industry has not been subject to FDA’s regulations, although there are projects being
implemented by the AFSS to change this. The AFSS also implements training, education, and
outreach activities to keep partners and stakeholders informed of FDA and state feed regulations
(Administration, 2016).
 The FDAAA requires FDA to establish an early warning system about unsafe pet food,
improve labeling for pet food (in development) and standards for pet food ingredients (also in
development). The agency requires timely information about unsafe animal feed and, where
appropriate, makes such information publicly available. In May 2010, FDA launched the Safety
Reporting Portal, where responsible parties can report foods to the registry. Consumers can also
report pet food complaints to the FDA through this registry. Additionally, vets can report
complaints on this registry due to pet illness they are treating. This allows FDA to know areas
requiring inspection. In March 2014, FDA also added a portal for reporting problems with
livestock feed; reports made for species considered to be farm animals (horse, cattle, swine,
poultry, fish)
The FSMA deals with prevention of food safety problems. FSMA gives FDA the legislative
power to require comprehensive, science-based preventive controls across the food supply,
including preventive controls for animal feed. The FDA enforcement authorities include:
mandatory recall authority, food safety records access, suspension of registration, administration
detention (designed to achieve higher rates of compliance), authority to ensure that imported
products meet U.S. standards are are safe for U.S. customers. All domestic and foreign human
food and animal feed facilities must register under the Public Health Security and Bioterrorism
Preparedness and Response Act of 2002 (Bioterrorism Act) (Administration, 2016).
Further constraints for creating pet foods include passing regulations created by AAFCO.
AAFCO deems pet food to be complete and balanced for pet diets and often require food testing
to determine the nature and composition of manufactured products (“Protein”, 2014).
Conclusion
Brewer’s spent grain is a valuable waste stream from the brewing process that has high
potential for further processing. Current methods of protein extraction involve the use of
chemicals or enzymes to break down fibers and proteins and enable the proteins to be separated
from the fiber portion of the spent grain. The BSG team aims to run separate experiments using
alcohol and enzymes to determine an achievable final purity of protein that can be recovered as
well as the amount of alcohol or enzymes that must be used. A final product high in protein and
low in fiber will increase the ability for both non-ruminant animals and humans to incorporate
into current diets. Using plate and frame filter presses, centrifuges, oven-drying apparatuses or
steam drying processes, sterilization procedures, lipid analyses, fiber analyses, and additional
experimental lab equipment, the team aims to understand the best practices and procedures
required to turn a waste product from the brewing industry into a high-value protein and biofuel.
Through experimentation the team will determine the costs and effects of using alcohol versus
the Alcalase enzyme in breaking down the brewer’s spent grain. As a team the various
experiments will be run and assessed using equipment in Haibo Huang’s food science lab on
Virginia Tech campus. The most significant considerations will be the amount of protein
recovered in small scale experimentation, the purity of the protein recovered, and the amount of
byproduct (fiber) collected by filter and frame press and centrifugation to be used for biofuel.
Determination of the economic value of the protein product and biofuel product will be
compared through multiple runs and following experimentation, a method will be chosen for
scale up using SuperPro designer software. The scale up of the team’s chosen method will
require processing data from a large scale brewery to simulate a real world design for breweries
of a certain size and processing volume. The team will contact local high volume breweries in
Roanoke to design a process that can be viable for breweries in the future to eliminate waste or
create high value products from current waste streams.
References

Dense Culture Food Pentair Available at: http://pentairaes.com/dense-culture-

lllllfood.html?gclid=EAIaIQobChMI6T56OCM1wIVXVgNCh0ZrwwiEAkYASABEgJszPD_Bl
lllllwE.

FSMA Compliance Solutions ORBCOMM. Available at:

lllllhttp://www2.orbcomm.com/fsmacompliance?gclid=EAIaIQobChMIxoHi1aGW1wIVxrrACh
lllll1IMQHKEAAYASAAEgJQSPD_BwE.

Quaker City Grinding Mills Quaker City Grinding Systems, LLC. Available at:
lllllhttp://www.qcgsys.com/.

VetterTec Spray Dryers VetterTec. Available at:

lllllhttp://www.vettertec.de/html/products/dryers/spray_dryers.html.

2014. Protein in Pet Food: What you Need to Know. Vet Street. Available at:
lllllhttp://www.vetstreet.com/protein-in-pet-food-what-you-really-need-to-know.

2017. Alcalase Enzyme, Bacillus licheniformis Millipore Sigma. Available at:

lllllhttp://www.sigmaaldrich.com/catalog/product/mm/126741?lang=en&region=US.

2017. Micro filtration and ultra filtration Lenntech Available at:

lllllhttps://www.lenntech.com/microfiltration-and-ultrafiltration.htm.

2017. Our Inventory. Kyte Centrifuge. Available at: http://kytecentrifuge.com/.

2017. Processing Vessels Available at: http://www.sprinkman.com/products/processing-systems.

2017. Water Filters and Water Treatment Systems Freshwater Systems Available at:
lllllhttps://www.freshwatersystems.com/.

2017. World population projected to reach 9.8 billion in 2050, and 11.2 billion in 2100. New
lllllYork: United Nations Department of Economic and Social Affairs. Available at:
lllllhttps://www.un.org/development/desa/en/news/population/world-population-prospects-
lllll2017.html.

Acton, Q. A. 2013. Hydroxides—Advances in Research and Application: 2013 Edition.

lllllScholarlyEditions. Administration, U. S. F. a. D. 2016. Overview of FDA's Animal Feed
lllllSafety System
Al-Hadithi, A., A. Muhsen, and A. Yaser. 1985. A study on the possibility of using some organic
lllllacids as preservatives for brewer's by products. Journal of Agriculture and Water Resources
lllllResearch (Iraq).

Aliyu, S., and M. Bala. 2011. Brewer’s spent grain: a review of its potentials and applications.
lllllAfrican Journal of Biotechnology 10(3):324-331.

Bartolomé, B., M. Santos, J. Jiménez, M. Del Nozal, and C. Gómez-Cordovés. 2002. Pentoses
llllland hydroxycinnamic acids in brewer's spent grain. Journal of Cereal Science 36(1):51-58.

Burden, D. W. 2012. Guide to the disruption of biological samples-2012. Random Primers 12(1).

Chhetri, G., P. Kalita, and T. Tripathi. 2015. An efficient protocol to enhance recombinant
lllllprotein expression using ethanol in Escherichia coli. MethodsX 2(Supplement C):385-391.

Crowe, N. L., I. Alli, and B. E. Baker. 1985. Solubilisation of Nitrogenous Constituents of

lllllbrewer’s spent grain. Journal of the Institute of Brewing 91(3):148-150.

Doncer, A. J., and H. R. White. 1975. Rotary vacuum filter. Google Patents.

Downing, A. L., and R. C. Squires. 1988. Cross-flow filtration. Google Patents.

Duong-Ly, K. C., and S. B. Gabelli. 2014. Salting out of proteins using ammonium sulfate
lllllprecipitation. Methods Enzymol 541:85-94.

El-Shafey, E., M. Gameiro, P. Correia, and J. De Carvalho. 2004. Dewatering of brewer's spent
lllllgrain using a membrane filter press: a pilot plant study. Separation Science and Technology
lllll39(14):3237-3261.

Faulds, C., A. Sancho, and B. Bartolomé. 2002. Mono-and dimeric ferulic acid release from
lllllbrewer's spent grain by fungal feruloyl esterases. Applied microbiology and biotechnology
lllll60(4):489-494.

Glass, K. A., L. M. McDonnell, R. C. Rassel, and K. L. Zierke. 2007. Controlling Listeria

lllllmonocytogenes on sliced ham and turkey products using benzoate, propionate, and sorbate. J
lllllFood Prot 70(10):2306-2312.
Iversen, C. 2015. Food Safety Best Practices in Food Transportation and the Role of ERP
lllllTechnology Available at: http://www.foodlogistics.com/article/12061249/food-safety-best-
lllllpractices-in-food-transportation-and-the-role-of-erp-technology.

Kissell, L., N. Prentice, and R. Lindsay. 1979. Protein and fiber enrichment of cookie flour with
lllllbrewers’ spent grain. Cereal Chem 56(4):261-266.

Kuo, K.-P., and M. Cheryan. 1983. Ultrafiltration of Acid Whey in a Spiral-Wound Unit: Effect
lllllof Operating Parameters on Membrane Fouling. Journal of Food Science 48(4):1113-1118.

Luche, S., V. Santoni, and T. Rabilloud. 2003. Evaluation of nonionic and zwitterionic
llllldetergents as membrane protein solubilizers in two-dimensional electrophoresis.
lllllPROTEOMICS 3(3):249-253.

Lynch, K. M., E. J. Steffen, and E. K. Arendt. 2016. Brewers' spent grain: a review with an
lllllemphasis on food and health. Journal of the Institute of Brewing 122(4):553-568.

Macheiner, D., B. F. Adamitsch, F. Karner, and W. A. Hampel. 2003. Pretreatment and

lllllHydrolysis of Brewer's Spent Grains. Engineering in Life Sciences 3(10):401-405.

Marinaccio, P. J., and R. V. Repetti. 1989. Cross-flow filtration. Google Patents.

Mathias, T. R. d. S., V. M. F. Alexandre, M. C. Cammarota, P. P. M. Mello, and E. F. C.

lllllSérvulo. 2015. Characterization and determination of brewer's solid wastes composition.
lllllJournal of the Institute of Brewing 121(3):400-404.

McCarthy, A. L., Y. C. O'Callaghan, C. O. Piggott, R. J. FitzGerald, and N. M. O'Brien. 2013.

lllllBrewers' spent grain; bioactivity of phenolic component, its role in animal nutrition and
lllllpotential for incorporation in functional foods: a review. Proceedings of the Nutrition Society
lllll72(1):117-125.

Mishra, P. K., T. Gregor, and R. Wimmer. 2016. Utilising Brewer’s Spent Grain as a Source of
lllllCellulose Nanofibres Following Separation of Protein-based Biomass. 2016 No. 1.

Mujumdar, A. S. 2006. Handbook of Industrial Drying, Third Edition. CRC Press.

Mussatto, S., G. Dragone, and I. Roberto. 2006. Brewers' spent grain: generation, characteristics
llllland potential applications. Journal of Cereal Science 43(1):1-14.
Nehete, J. Y., R. S. Bhambar, M. R. Narkhede, and S. R. Gawali. 2013. Natural proteins:
lllllSources, isolation, characterization and applications. Pharmacognosy Reviews 7(14):107-116.

Oksman, K., A. P. Mathew, A. Bismarck, O. Rojas, and M. Sain. 2014. Handbook of Green
lllllMaterials: Processing Technologies, Properties and Applications (in 4 volumes). World
lllllScientific.

Perry, E. 2017. Pressing Spent Brewers Grains to improve its use as alternative feed: A Study of
lllllits effect on Dairy Sheep and Meat lambs. FNE09-656. U. SARE.

S, B. V. 1965. Recovery of edible products from spent grains and yeasts. Google Patents.
lllllSantos, M., J. Jiménez, B. Bartolomé, C. Gómez-Cordovés, and M. Del Nozal. 2003.
lllllVariability of brewer’s spent grain within a brewery. Food Chemistry 80(1):17-21.

Sežun, M., V. Grilc, G. D. Zupančič, and R. M. Logar. 2011. Anaerobic Digestion of Brewery
lllllSpent Grain in a Semi-Continuous Bioreactor: Inhibition by Phenolic Degradation Products.
lllllActa Chimica Slovenica 58(1):158-166.

Spence, K. L., R. A. Venditti, O. J. Rojas, Y. Habibi, and J. J. Pawlak. 2011. A comparative

lllllstudy of energy consumption and physical properties of microfibrillated cellulose produced
lllllby different processing methods. Cellulose 18(4):1097-1111.

Ström, L.-L. Evaluation of Pilot Scale Drying of Brewer’s Spent Grain in a Keith Rotary
lllllSuperheated Steam Dryer-a Preliminary Investigation.

Tang, D.-S., G.-M. Yin, Y.-Z. He, S.-Q. Hu, B. Li, L. Li, H.-L. Liang, and D. Borthakur. 2009.
lllllRecovery of protein from brewer's spent grain by ultrafiltration. Biochemical Engineering
lllllJournal 48(1):1-5.

Treimo, J., S. I. Aspmo, V. G. H. Eijsink, and S. J. Horn. 2008. Enzymatic Solubilization of

lllllProteins in Brewer’s Spent Grain. Journal of Agricultural and Food Chemistry 56(13):5359-
lllll5365.

Walter, J. 1970. Plate for plate and frame filter presses. Google Patents. Witkiewicz, K. 2012.
lllllSustainable Uses of Spent Grain. Craft Beer Muses. Available at:
lllllhttps://www.craftbeer.com/craft-beer-muses/sustainable-uses-of-spent-grain.
 Appendix
Appendix 1: Brainstorming
When the team first met up and discussed the project, groups targeted the final brewers
spent grain product to animal feed. The team debated what consumer market was desirable to
target and what economic incentives were behind each market. Ideally the group would like to
produce a protein to alleviate the protein demand of humans by forming a high purity protein
powder to be used as and ingredient or additive to recipes. The team also considered the
possibility of different food items that this high purity, being 80-90% pure, could be turned into.
Next the team debated using the protein to feed fish and other animals such as cats, dogs, and
non-ruminant animals.
The second meeting was more focused on the chemical components of the BSG. The
team summarized the first two discussions on BSE senior presentation PowerPoint.
After thinking to be more creative and not limited to the single member's thought, the team
separated 4 members into two group based on the specific target. Zhicheng Xu and Ryan
Mitchell were the teams specific to pet (cat or dog) feed research. Samuel Elizondo and Lucy
Epshteyn were tasked with searching for more information about the fish feed production.
Due to the complexity of current chemical and biological processes required to convert
brewers spent grains into a human worthy high purity protein that is 80-90% pure, the team
instead decided to produce a feed with the hopes of obtaining 30-50% protein purity. This allows
the team to use the final protein as a fish or animal food feed. The market for fish food is high as
farm raised fish is ever increasing in popularity, and the pet food market can be targeted for as a
high-value feed market.
Along with creating a feed for fish or animals that is valuable and marketable, the team
also aims to use the fiber that is filtered out of the finished feed product as a biofuel. The fiber
that is high in cellulose can be used for burning in pellet stoves, and pellet processing machines
can be easily purchased and run to create high value fuels. The team aims to find a lab on
Virginia Tech campus that will allow for the determining of the energy value of the fibrous
material that is leftover and removed from the feed product.
 After multiple discussions between the team and Haibo Huang, the team’s academic
advisor, a decision was made to use experimental equipment in the lab to determine how much
protein can be extracted from the brewer’s spent grain through simpler methods. The team will
use various equipment to take various lipid, fiber, and protein panels, and also determine the
energy value of fibrous materials. Following a series of varied experimental runs and analyzing
differences among the runs, the team will use SuperPro designer software to scale up the most
feasible and economically viable method.
Appendix 2: Challenges Encountered and Plans to Address
The major challenge the team will overcome is the economic extraction of protein with a
method that is according to the food and feed standards imposed by the FDA. There are two
main methods of extraction. One method is via alcohol degradation of the carbohydrate layer of
the BSG, this method is cheap and in theory efficient as most of the alcohol inputted can be
recovered and reutilized via distillation and ultrafiltration. The application of hazardous
chemicals in food and feed is highly regulated by the FDA. After the extraction of protein via the
alcohol method there would be numerous tests that the final product would have to overcome in
order to meet standard FDA guidelines. Sampling and standardized testing of the final product
will potentially add lag time to our process and increase fixed costs. The second method of
protein extraction of BSG is via enzyme hydrolysis. In theory and proven with research, up to
77% of the protein can be recovered from BSG with the single use of the enzyme Alcalase. The
major constraint with protein extraction via enzyme degradation is the increased fixed capital
investment to purchase and run the unit operations required to produce a finished product.
Market available enzymes are expensive and require special conditions to optimize functionality.
Extensive economic analysis and predictive analytics must be formed and evaluated to finalize
the ideal protein extraction method.
Transportation will also be a key challenge that our final design will have to overcome.
Fresh BSG has a high water and moisture content and is rich in sugars, providing ideal
conditions for microorganisms and fungi to grow and prosper. Ideally the BSG would be dried
right immediately following extraction from the brewing fermenters to protect the spent grains
from any microorganism growth. The spent grains would then be transported under special
conditions to minimize exposure. The BSG group will have to work closely with the breweries
and invest in novel transportation technology to mitigate risk from this process.
The last key challenge is that BSG composition varies from each brewery and each batch
of beer produced. It will be hard to standardize a uniform amount of expected protein to be
extracted from each run. Marketing a protein product will have difficulties considering the vast
market of proteins in existences today, however the health benefits behind plant based proteins
enable brewer’s spent grains to be viewed as a healthy plant protein or meat product alternative.
An extensive marketing campaign will likely have to be launched to inform the general public of
the advantages of our product and its environmental benefits.
Appendix 3 - Design Ideas
Figure 1: Protein Extraction with Alcohol
Figure 2: Protein Extraction with Enzyme

Figure 3: Team Brainstorming

Appendix 4: Gantt Chart
Appendix 5: Team Member Responsibilities
The team split the tasks required to complete the technology review into categories
shown in the table below. Smaller tasks were divided evenly among the group. The bulk majority
of the technology review was completed by all members who researched different areas of
protein separation and brewer’s spent grain.

Task Completed Member Responsible

Cover Letter Ryan
Cover Page Lucy
Introduction Samuel, Lucy, Ryan
Technology Review Zhicheng, Samuel, Lucy, Ryan
Conclusions Ryan
References Lucy
Team Brainstorming Zhicheng
Gantt Chart Samuel
Team Challenges Samuel