NSERC funding application- Example 2

1. Soheila Abachi Hokmabadi Nazhad
Of all the food produced in 2009, globally, 32% (by weight) was lost and wasted equaling
for 24% loss of the total calories (FAO). World fishery production in 2006 was approximately 143.6
million tones by major fishing countries of which 76.9% was utilized for direct human consumption
(Nguyen, Sylla et al. 2011). Quantitatively, the food loss and waste for fish and seafood is around
35% (FAO). By-products of the fishery processing mainly become the raw material for production
of feed, fishmeal, oil, protein isolates and hydrolysates. In a time that food security, sustainability
and environmental protection are major concerns of the century, worldwide, utilization of food
processing plants’ by-products has to be the most prominent breakthrough of all. In addition,
chronic cardiometabolic diseases, mainly representing diabetes, cardiovascular disease, and obesity,
are predicted to be one of the most prominent public health concerns in the next decade or so.
Statistics point up that these conditions impose great economic burden on the societies worldwide
and are of great importance as the related mortality rates are significant. Cardiovascular diseases are
the leading cause of death worldwide claiming the life of 17.5 million people accounting for 31% of
the total deaths in 2012 (WHO). The potential of food-derived, plant or animal, bioactive peptides
in decreasing the risk of cardiovascular diseases have extensively been reviewed (Erdmann, Cheung
et al. 2008).
Bioactive peptides of animal source can be extracted mainly from milk, egg, meat and fish
of various types including pelagic fish (Erdmann, Cheung et al. 2008). Statistics by Fisheries and
Oceans Canada show that in 2014, Atlantic and Pacific coast commercial landings for groundfish,
pelagic and other finfish were 358,712 metric tons (live weight) mackerel accounting for 6,540
metric tons. To separate the primary and secondary metabolites of the fishery by-products
enzymatic hydrolysis has been the most promising method among other bio-techniques (Beaulieu,
Thibodeau et al. 2009; Chalamaiah, Hemalatha et al. 2012). Membrane filtration, chromatography
or electromembrane filtration could isolate peptides from protein hydrolysates however latter is the
preferred method of the separation for strongly charged molecules with enhanced selectivity and
cost efficiency (Bargeman, Koops et al. 2002). Based on chemical structure, marine metabolites
have been categorized into alkaloids, terpenes, aethers/ketals, strides, lactones,
hrydroxybenzenes/chinones and peptides nevertheless, statistically, peptides have the highest
proportion of bioactivity, 40.85%, among all (Hu, Chen et al. 2015). Studies have shown the
beneficiary properties of cod fish dietary protein on insulin sensitivity and glucose tolerance in
sucrose-fed mice, in-vivo (Lavigne, Marette et al. 2000; Tremblay, Lavigne et al. 2003). Salmon
fish protein containing high fat/sucrose diets notably prevent the weight gain in mice (Pilon, Ruzzin
et al. 2011). Herring, mackerel, and salmon fish protein hydrolysate diets moreover exhibit anti-
inflammatory attributes through modulation of tumor necrosis factor–α and interleukin-6 expression
levels, in visceral adipose tissue (Pilon, Ruzzin et al. 2011).
In the proposed research work, we will focus on the mackerel by-products as model. We
hypothesize that mackerel by-products’ bioactive-peptide fraction(s) can modulate glucose uptake
and demonstrate anti-inflammatory activity in lipopolysaccharide (LPS) treated macrophages, in-
vitro. In this investigation, we aim to fractionate, characterize and identify the bioactive peptide(s)
with promising glucose uptake modulation and anti-inflammatory activity in LPS treated
macrophages. In this study, we will fractionate and purify the fractions of molecular size of 0.2-10
kDa using membrane technology (Beaulieu, Thibodeau et al. 2009; Suwal 2015) and these fractions
will be successively fractionated into subtractions using electrodialysis with ultrafiltration
membrane (Chevrier, Mitchell et al. 2015). Sequentially the effect of parameters; pH, pore size, cell
configuration on the performance of electrodialysis with filtration membrane in fractionation of our
molecules of interest will be evaluated (Suwal 2015). The fractions will be screened for anti-
inflammatory (Roblet, Doyen et al. 2013) and glucose uptake modulation effect, in-vitro (Pilon,
Ruzzin et al. 2011; Roblet, Akhtar et al. 2016). Simultaneously, the fractions will be characterized
using LC-MS/MS: ESI-Q-TOF (Doyen 2011; Suwal 2015).

2. Soheila Abachi Hokmabadi Nazhad
Bargeman, G., G. Koops, et al. (2002). "The development of electro-membrane filtration for the
isolation of bioactive peptides: the effect of membrane selection and operating parameters on the
transport rate." Desalination 149(1): 369-374.
Beaulieu, L., J. Thibodeau, et al. (2009). "Proteolytic processing of Atlantic mackerel (Scomber
scombrus) and biochemical characterisation of hydrolysates." International journal of food
science & technology 44(8): 1609-1618.
Chalamaiah, M., R. Hemalatha, et al. (2012). "Fish protein hydrolysates: proximate composition,
amino acid composition, antioxidant activities and applications: a review." Food chemistry
135(4): 3020-3038.
Chevrier, G., P. L. Mitchell, et al. (2015). "Low-molecular-weight peptides from salmon protein
prevent obesity-linked glucose intolerance, inflammation, and dyslipidemia in
LDLR−/−/ApoB100/100 mice." The Journal of Nutrition 145(7): 1415-1422.
Doyen, A. (2011). Fractionnement d'un hydrolysat peptidique de co-produits de crabe des neiges
par électrodialyse avec membranes d'ultrafiltration: impact des paramètres liés au procédé sur la
migration et la sélectivité peptidique, Université Laval.
Erdmann, K., B. W. Cheung, et al. (2008). "The possible roles of food-derived bioactive peptides in
reducing the risk of cardiovascular disease." The Journal of nutritional biochemistry 19(10): 643-
654.
FAO "Available online: http://www.fao.org/save-food/resources/keyfindings/en/ (accessed on 29 09
2016)."
Hu, Y., J. Chen, et al. (2015). "Statistical research on the bioactivity of new marine natural products
discovered during the 28 years from 1985 to 2012." Marine drugs 13(1): 202-221.
Lavigne, C., A. Marette, et al. (2000). "Cod and soy proteins compared with casein improve glucose
tolerance and insulin sensitivity in rats." American Journal of Physiology-Endocrinology And
Metabolism 278(3): E491-E500.
Nguyen, H. T. M., K. S. B. Sylla, et al. (2011). "Enzymatic hydrolysis of yellowfin tuna (Thunnus
albacares) by-products using Protamex protease." Food Technology and Biotechnology 49(1):
48-55.
Pilon, G., J. Ruzzin, et al. (2011). "Differential effects of various fish proteins in altering body
weight, adiposity, inflammatory status, and insulin sensitivity in high-fat–fed rats." Metabolism
60(8): 1122-1130.
Roblet, C., M. J. Akhtar, et al. (2016). "Enhancement of glucose uptake in muscular cell by peptide
fractions separated by electrodialysis with filtration membrane from salmon frame protein
hydrolysate." Journal of Functional Foods 22: 337-346.
Roblet, C., A. Doyen, et al. (2013). "Impact of pH on ultrafiltration membrane selectivity during
electrodialysis with ultrafiltration membrane (EDUF) purification of soy peptides from a
complex matrix." Journal of membrane science 435: 207-217.
Suwal, S. (2015). Fractionation of Peptides from Protein Hydrolysate by Electrodialysis with
Filtration Membrane: Process Optimization, Fouling Characterization and Control Mechanisms,
Université Laval.
Tremblay, F., C. Lavigne, et al. (2003). "Dietary cod protein restores insulin-induced activation of
phosphatidylinositol 3-kinase/Akt and GLUT4 translocation to the T-tubules in skeletal muscle
of high-fat-fed obese rats." Diabetes 52(1): 29-37.
WHO "Available online: http://www.who.int/mediacentre/factsheets/fs317/en/ (accessed on 29 09
2016)."