Fish protein hydrolysate

04-13-16 Fish protein and its effects on body weight 1

With the development of modern society, prevalence of
overweight and obesity is seriously threatening global
health.
Main reasons are people’s life style of decreased physical
activity and intake of high energy food.
Overweight and obesity are risk factors for several diseases
and disorders, and the most common are insulin resistance,
type 2 diabetes mellitus, high blood pressure, dyslipidemia
and CHD.
Not only the quantity of food intake and also composition
of diet may be important for achieving and maintaining
weight loss over time.
A modest increase in protein intake combined with a
modest reduction in glycaemic index shows positive
results.

In the U.S., 70.9 percent of men and 61.9
percent of women are overweight or obese,
compared to 38 percent of men and 36.9
percent of women worldwide.
Source: Lancet Medical Journal 2014
28.8 percent of boys and 29.7 percent of girls
are overweight or obese in the U.S., compared
to 14.2 percent of boys and 14.7 percent of
girls worldwide.

Most medical societies, recommend a diet relatively rich in carbohydrate and low
in fat to prevent obesity and related diseases.
However, the public interest in alternative diets is increasing, in particular for diets
with high protein and high fat content in relation to carbohydrates.
The long-term regulation of maintenance of body weight is integrated by endocrine
signals and coupled with short-term signals related to meal size, like
cholecystokinin (CCK) and GLP-1.
This Contributes to regulate daily energy intake in
synchronization with long-term body weight
regulation.
High intake of dietary protein at the expense of
carbohydrates increases satiety in the short term,
and may thereby reduce the intake of energy.

Protein Hydrolysates are able to stimulate CCK and GLP-1 secretions and their respective
gene transcription in stomach.
Protein confers the highest satiating effect of all macronutrients.
Because of the strong potential of protein to stimulate satietogenic peptides secretion, their
utilization as appetite suppressive products in the prevention and treatment of obesity has
evidenced growing interest last few years.

Fish Protein hydrolysates (FPHs) are complex mixtures of free amino acids and
small peptide fragments.
Obtained by breaking down naturally occurring intact proteins from by-products
of fish, which includes skin, head, viscera, trimmings, liver, frames and bones.
It is a breakdown product of enzymatic conversion of fish proteins into smaller
peptides, which normally contain 2–20 amino acids. It contains minimum 70%
of hydrolyzed protein.
604-13-16
Fish protein Production
Fish protein and its effects on body weight

Fish protein hydrolysates (FPHs) have been reported as a suitable source of
proteins for human nutrition because of their balanced amino acid
composition and positive effect on gastrointestinal absorption.
Studies on fish-derived peptides demonstrated antihypertensive, antioxidant,
immunomodulating effects, reparative properties in the intestine, and effects
in reducing plasma cholesterol and triglycerides levels.
Protein hydrolysates can be produced using a variety of methods, including
heating with acids or by enzymatic treatment with either endogenous or
exogenous proteolytic enzymes.
Liquid Fish protein Powder Fish protein

Nobile, V., Duclos, E., Michelotti, A., Bizzaro, G., Negro, M., & Soisson, F. (2016).

Investigated the effect of a fish protein hydrolysate, obtained from blue
whiting muscle by enzymatic hydrolysis, on body composition and on
stimulating cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1)
secretion.

A randomized clinical study was carried out.
Total Population – 120n
Female – 90n
Male – 30n
Slightly overweight (25 kg/m2 ≤ BMI<30 kg/m2)
Age – 18 to 55 years
Study Period – 90 days

Body composition measurements:
Body weight and height were measured using
electronic balance and an upright stadiometer.
Fat mass was measured using Dual Energy X-
ray Absorptiometry.
Extracellular water was measured in the supine
position, using BIA technique.
Circumference of waist, hips, and thighs (left
and right) were measured using a flexible
measuring tape (precision: 1 mm).
04-13-16 Fish protein and its effects on body weight 13X-ray
BIA
Weighing
Scale
Stadiometer

Biochemical analysis:
Total CCK and GLP-1 levels in fasting blood plasma
were assessed using commercially available ELISA
(enzyme-linked immunosorbent assay).

Statistical Analysis:
Statistical analysis was performed using NCSS 8 statistical software.
Data normality was checked using Shapiro–Wilk normality test and data shape.
Intragroup and intergroup comparisons were carried out using repeated measures
analysis of variance followed by Tukey–Kramer post-test.
A p-value <0.05 was considered statistically significant.
Intragroup statistical analysis was carried out on raw data, whereas
intergroup statistical analysis was carried out on the changes from
baseline.

CCK and GLP-1 blood levels
Intragroup (vs. D0) statistical analysis is reported upon the bars of the histogram. The lines report the
intergroup (vs. placebo) statistical analysis. Statistical analysis is reported as follows: *p<0.05, **p<0.01, and
***p<0.001. Data are mean±SE. Fish protein and its effects on body weight 17

Both the 1.4 and 2.8 g FPH dose increased the serum levels of CCK and GLP-1.
The increase of CCK and GLP-1 was statistically significant when compared with placebo
without any differences among the two tested doses after 90 days of treatment.
Body composition was changed. Body weight, BMI, fat mass, and circumferences were
statistically significantly decreased when compared with placebo after 90 days of treatment.
Notably, the decrease in body weight was not related to extracellular water decrease
(‘draining effect’) but was related to fat mass decrease (‘slimming effect’).
No differences in CCK/GLP-1 blood concentrations and changes in markers of body
composition were found between the 1.4 and 2.8 g concentration, indicating a plateau effect
starting from the 1.4 g FPH doses.
The putative mechanism of action of FPH in body weight management could be as follows:
1) an increase of CCK and GLP-1 blood levels, 2) a decrease in food intake (meal size)
stimulated by CCK and GLP-1 secretion in the short term, and 3) a decrease of adipose tissue
mass mediated by GLP-1 interaction with leptin in the long term. 18
04-13-16

Strengths Limitations
Results were analyzed with well known
statistical tools.
Mechanism of action of the FPH was not
explained.
Risk of bias was minimum due to RCT. There were dropouts in the study which
effected the population size.
Population size was good enough.
Dose effect relationship was measured.
Excluded subjects using food supplements
containing ingredients having influence on
body weight.
Fish protein and its effects on body weight04-13-16 19

Fish protein by enzymatic hydrolysis helps in stimulating the secretion
of CCK and GLP-1 and was effective on body weight and particularly
in controlling body composition.

Type of Fish used as a protein source in Study-1?
a. Cod Fish
b. Blue Whiting
c. Salmon Fish
b

Vikøren, L. A., Nygård, O. K., Lied, E., Rostrup, E., & Gudbrandsen, O. A. (2012).

Investigated the effect of supplementation with a low daily dose of fish
protein on glucose tolerance, blood lipids, blood pressure and body weight
in overweight and otherwise healthy adults.

A double-blind, randomized, controlled intervention study.
Total Population – 40n
Female – 20n
Male – 20n
BMI ≥ 27 kg/m2
Fasting blood glucose < 7 mmol/l
Age – 20 to 70 years
Study period – 8 weeks

Body composition measurements:
Body composition were measured using a
bioelectrical impedance analysis device (BIA).
Blood pressure was measured using the Sciller
BP-200plus.
Body weight and height were measured.
Blood samples were collected for isolation of
serum.
The macronutrient and energy content was
calculated using ‘Mat pa˚ Data 5.1’.
Weighing
Scale
BIA
Stadiometer

Insulin sensitivity (Homeostasis Model Assessment –
Insulin Resistance (HOMA-IR)):
The HOMA-IR model was used to evaluate insulin
sensitivity in the present study, estimated by
mathematical modelling of fasting glucose and fasting
insulin concentrations.
It was calculated from fasting plasma glucose and
insulin concentrations as glucose (mmol/l) × insulin
(mU/l) divided by 22·5.
04-13-16

Statistical Analysis:
Statistical analysis was performed using PASW Statistics 19.0 software.
Data normality was checked using Shapiro Wilk normality test and data shape.
The majority of data were not normally distributed and therefore non-parametric tests
were used for all data comparisons.
The Mann Whitney U test was used to compare data between groups.
The Wilcoxon signed ranks test for paired samples was used to compare changes
within groups over the time course of the study.
Data are presented as medians with interquartile ranges.
P values,0.05 were considered statistically significant.

04-13-16 32Fish protein and its effects on body weight

No statistically significant differences were found between the treatment groups for
age or BMI.
No statistically significant changes were seen in the macronutrient and energy intake
during the course of the study.
Based on the interviews, there were no significant changes in physical activity in
either group.
At the end of the study, the fasting glucose level was significantly lower in the fish
protein group when compared to placebo, whereas no differences were seen in
fasting insulin, fasting insulin C-peptide and HOMA-IR at this time point.
AUC for five glucose measurements was significantly improved after 8 weeks, and
fasting glucose.
A significant reduction in postprandial insulin C-peptide levels were observed after 8
weeks in the fish protein group. 28
04-13-16

Serum levels of total cholesterol and HDL-cholesterol were not significantly
different between groups at any of the three time points.
The LDL-cholesterol level was significantly reduced in the fish protein group at
the end of the study compared to baseline whereas no change was seen in the
placebo group.
A significant increase in the HDL:LDL ratio was seen in the fish protein group at
the end of the study when compared to baseline.
No significant differences were seen for systolic or diastolic blood pressure
between the fish protein group and control group.
Body weight and percentage of muscle were significantly increased during the
first 4 weeks of the study in the fish protein group, concomitant with a significant
reduction of percentage of body fat.
29
04-13-16

Strengths Limitations
Data collection methods were accurate. Mechanisms by which codfish proteins
improve glucose tolerance are unclear.
Results were analyzed with well known
statistical tools.
Subjects were recruited through adverts in
the newspaper.
Statistically nonsignificant result may be due
to insufficient Subjects.
Study period was very short.

A daily consumption of fish protein supplement without changing the
dietary regimen and lifestyle may be sufficient to decrease LDL
cholesterol and improve glucose tolerance and body composition in
overweight adults, but did not affect blood pressure, HDL-cholesterol,
bile acids, C-Reactive Protein (CRP).

Total no of subjects participated in Study-2?
a. 40
b. 80
c. 120
a

Study 1 Study 2
Effect of FPH supplementation on body composition and
gut hormones secretions in adults.
Effect of low daily dose FP supplementation on glucose
tolerance, lipids and body composition in adults.
Randomized clinical study Double-blind, randomized, controlled intervention study
Body composition measurements (weight, height and X-
ray) and Biochemical analysis (ELISA).
Body composition measurements (weight, height and
blood pressure) and HOMA-IR.
Statistical Analysis: Comparisons were done using
repeated measure analysis of variance, Tukey–Kramer
post-test and Shapiro–Wilk normality test and data shape.
P-value <0.05 was considered statistically significant.
Statistical Analysis: Mann Whitney U test for comparing
groups, Wilcoxon signed ranks test for paired samples to
compare changes in group and Shapiro Wilk normality
test and data shape.
CCK and GLP-1 blood levels were increased which
facilitates intake of small quantity of food and effective
in maintaining body composition.
Daily low consumption of FP helps in improving glucose
tolerance and body composition in adults.
Strengths: Dose effect relationship, Sample size, RCT Strengths: Data collection methods and statistical
analysis
Limitations: Mechanism action of FPH unclear, Dropouts Limitations: Small no of subjects, short period.

Fish protein isolated from inexpensive fish could provide a new basis for the
incorporation of omega-3 polyunsaturated fatty acids in to functional foods.
It is a logical vehicle for increasing the consumption of omega-3 PUFAs, production of
gut hormones associated with suppressing appetite without the need for dietary
supplements in a pill or capsule form.
Functional Foods “ are food products that contain added, technologically developed
ingredients with specific health benefits.”
Standardized procedures to examine the functional properties are needed, as well as
more studies on using endogenous enzymes to make functional FPH.
Moreover, in the global context of marine resource depletion it could be of economical
and ecological interest to promote the development of fish by-products upgrading.

Nobile, V., Duclos, E., Michelotti, A., Bizzaro, G., Negro, M., & Soisson, F. (2016). Supplementation with a fish protein hydrolysate (
Micromesistius poutassou ): Effects on body weight, body composition, and CCK/GLP-1 secretion. Food & Nutrition Research, 60(0).
doi:10.3402/fnr.v60.29857
Vikøren, L. A., Nygård, O. K., Lied, E., Rostrup, E., & Gudbrandsen, O. A. (2012). A randomised study on the effects of fish protein
supplement on glucose tolerance, lipids and body composition in overweight adults. British Journal of Nutrition Br J Nutr, 109(04), 648-657.
doi:10.1017/s0007114512001717
Liu, L., Wang, Y., Peng, C., & Wang, J. (2013). Optimization of the Preparation of Fish Protein Anti-Obesity Hydrolysates Using
Response Surface Methodology. IJMS International Journal of Molecular Sciences, 14(2), 3124-3139. doi:10.3390/ijms14023124
Cudennec, B., Fouchereau-Peron, M., Ferry, F., Duclos, E., & Ravallec, R. (2012). In vitro and in vivo evidence for a satiating effect of
fish protein hydrolysate obtained from blue whiting (Micromesistius poutassou) muscle. Journal of Functional Foods, 4(1), 271-277.
doi:10.1016/j.jff.2011.12.003
Nesse, K. O., Nagalakshmi, A., Marimuthu, P., Singh, M., Bhetariya, P. J., Ho, M., & Simon, R. R. (2014). Safety evaluation of fish
protein hydrolysate supplementation in malnourished children. Regulatory Toxicology and Pharmacology, 69(1), 1-6.
doi:10.1016/j.yrtph.2014.02.009
Gálvez, R. P., & Bergé, J. (n.d.). Utilization of fish waste.
Fish protein could provide basis for new wave of functional foods, suggest researchers. (n.d.). Retrieved April 13, 2016, from
http://www.nutraingredients.com/Research/Fish-protein-could-provide-basis-for-new-wave-of-functional-foods-suggest-
researchers?utm_source=copyright
Drammatic Organic Fertilizer. (n.d.). Retrieved April 13, 2016, from http://www.dramm.com/html/main.isx?sitesec=3.0.0.0
Yogurt Nutrition Digest 3, Ep. 2: Impact on satiety of macronutrients - Yogurt in Nutrition. (2015). Retrieved April 13, 2016, from
http://www.yogurtinnutrition.com/yogurt-nutrition-digest-3-ep-2-impact-on-satiety-of-macronutrients/

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