FUNCTIONAL BIOACTIVE PEPTIDES FROM SELECTED

AQUATIC SOURCES

Niche areaFOOD SECURITY

Prepared by: Prof. Abdul Salam Babji Assoc. Prof. Dr. Amiza Mat Amin Assoc Prof. Dr. Alina Abdul Rahim Assoc. Prof. Dr. Wan Rosli Wan Ishak Assoc Prof. Dr. Noriham Abdullah Mr. Ismail Ishak & Dr. Wan Norhana

Bioactive Peptides• Biologically active peptides play an important role in metabolic regulation

and modulation.

• These peptides can be used as functional food ingredients or nutraceuticals and pharmaceuticals to improve human health and prevent diseases.

• The importance of fish as a source of novel bioactive substances is growing significantly (Aneiros and Garateix, 2004; Barrow and Shahidi, 2008).

• Based on previous studies, bioactive peptides from fish are economically useful functional ingredients in food, food supplements and the pharmaceutical industry.

Aquatic Sources

• Fish meat – a major protein food group, with excellent sources of some valuable nutrients

• With red meat being associated with negative image related to health, fish based protein become the better choice.

Background studies• Najafian and Babji (2012) extensively reviewed fish derived antioxidant and

antimicrobial peptides – their production, assessment and applications.

• Recent studies by Ghassem et al. (2011), Ghassem et al. (2012), Mahmoodani et al. (2012) and See et al. (2010), all searching on the fundamentals of fish-based hydrolyzed proteins (peptides) have indicated functional properties associated toward benefits to mankind .

• Bioactive peptides involved include:– antihypertensive (ACE inhibitor), – anti-oxidative, – antimicrobial – economic production of collagen and gelatin from fish and waste from

the processing of fish.

Research Expectation

• The impact of research on functional peptides from fish proteins is expected to benefit the country’s fish industry by :1. economic, and environmental benefits

• Improving self sufficiency of fish supply : less dependence on imported oceanic fish

• Sustainability of freshwater aquaculture• Conservation of wild species of fish and preserving

biodiversity

Research Benefits2.Increase the potential production of health-related

functional ingredients and bioactive enriched food • suitable Halal compliant alternatives in food ingredients

and products from fish, as alternative choices against haram health products already available in the functional food markets such as:

– ACE inhibitory peptides of pork which decrease antihypertensive activity

– Pork antioxidative, antistress, antifatigue hydrolysates – Porcine based prebiotic peptides

• Discourage usage of using non-Halal innards of the livestock industry as fish feed to the freshwater fish in commercial farms, thus ensuring Halal food throughout the whole supply chain

Uses/ Applications of Bioactive Peptides

• The freeze-dried samples can be added to the spice used for garnishing.

• The freeze–dried samples can be used as food supplements.

• Selective purified peptides can be used as nutraceutical

• Fine grade peptides can be used for pharmaceutical• Immunostimulant properties of peptides to fish.

OPPORTUNITIES AND INNOVATIVE CHALLENGES

• Novel peptides from fish – Sensory properties

• Innovative? Acceptable? What do modified peptides taste like?

– Toxicity• Are new peptides toxic? Allergen?

– Nutrition• Bioavailability and specific target cells for enhanced absorption

– New functional food ingredients• Look into new ways of using by-products from the fish industry

as Halal alternative edible food ingredients– Enzymes, gelling agents, emulsifiers etc

Research Program Description

• 1. UKM Research Team will lead this project with:

• Technological processes for production of bioactive peptides from freshwater fishes.

• Analytical method and Bioassay tests of specific peptides.

• Pilot plant production studies.

Research Program Description

• 2. UMT Expertise: • Fermentation/hydrolysis process for

production of protein hydrolysate and bioactive peptides from marine-based sources

• Utilization of the marine-based hydrolysate and bioactive peptides in food and nutraceutical products

Research Program Description

• 3. USIM Expertise: • Desirable and unacceptable off-flavours formation• Products development shelf life studies as food

ingredients and health foods• Chemometrics usage studies on shelf life and flavours• 4. USM (Kubang Kerian) Expertise:• Food (Fish) processing• Fish products development

Research Program Description

• 5. UITM (Shah Alam) Expertise:• Comparative antioxidant and antimicrobial activities of

marine bioactive peptides to plants hydrolysates (extracts) • Toxicological testing of marine bioactive peptides in vivo

and in vitro

• 6. FRI Expertise:• Basic science on fish and fish products• Fish information, statistics and link with DOF and MOA.• Scale-up processing of fish and fish products.

Research Methodology

Basic generation method for peptides from food proteins

Typical procedure of isolation of functional bioactive peptides from fish meat proteins

Sarcoplasmic and myofibrillar powder processing

Flowchart of Extraction of Bioactive Peptide (Angiotensin Converting Enzyme (ACE) Inhibitors) of Sarcoplasmic and Myofibrillar Protein From Fish Meat

Fish skin collagen/gelatin powder processing

In processing chamber:

Step 1: Raw material

preparation Step 2: Cleaning

process Step 3: Alkaline pre-

treatment

Step 4: Acidic pre- treatment

Step 5: Collagen/ Gelatin extraction

Step 6: Purification and freeze dried.

Cold Water In

Water (cooling) medium

Water/ salt/

alkaline/ acidic

solution in

Temperature Indicator(5 °C)

Fish skin

Production of Bioactive Peptides

2% enzyme is added into the mixture.

Hydrolysis of sample by constant stirring (200rpm) at 55 °C is carried out for 2 hours.

Purification of bioactive hydrolysates for further analysis.

Water (heating) medium

pH Indicator(pH 7)

Temperature Indicator(55 °C)

sample

2% enzyme

in

Steam In

Evidence of Strength of Research Group

• Abdul Salam Babji (Leader): UKM– More than 30 years experiences in process meat products research. H

index 10 (daging@ukm.my)

• Amiza Mat Amin: UMT– 18 years experiences in food protein research. H index 3 (

ama@umt.edu.my)

• Alina Abdul Rahim: USIM– 10 years experiences in research. H index 3 (alinarahim@gmail.com); (

alina@usim.edu.my)

Evidence of Strength of Research Group

• Wan Rosli Wan Ishak: USM– 10 years experiences in research. H index 3 (rosliishak@gmail.com)

• Noriham Abdullah: UITM– 10 years experiences in functional food research. H index 3 (

noriham985@salam.uitm.edu.my)

• Ismail Hashim + Dr. Wan Norhana: Fishery Research Institute– More than 30 years experience in fisheries sciences especially in

environmental and food safety aspects (ismail4852@gmail.com)

R&D Products from Fisheries 2010 (FRI-IPTA)

• Research on freshwater fish (patin, keli and haruan) for identification of bioactive component have been conducted. – Lead by Prof. Dr. Abd. Salam Babji (UKM). – Yield on identified antihypertensive peptides from fish protein which

can replace captopril for hypertension treatment.

Hydrolysed collagen from keli skin

Collagen drink with flavors

Keli paste

Keli fishball

Haruan fishball

R&D Products from Fisheries 2010 (IPP-IPTA)

• Research on production of trypsin enzyme and hydrolysate have been conducted.– Lead by Assoc. Prof. Dr. Amiza Mat Amin (UMT). – The enzyme was extracted from wastes of (lake haruan and patin)

stomach’s part. Hydrolysates also was produced from the wastes of industrial fish processing which include stomach and bones.

Fish’s stomach for enzyme processing

Expected Output

This project will be undertaken for 4 years:• No. of Post-Doc. = 6 (2 from UKM and 1 from

each of other universities)• No. of PhD students = 10 (2 from each universities)• No. of MSc. students = 20 (4 from each universities)• No. of publication = 32-35• New technologies = 2-3• New fish-based products= 5-9• New food ingredients = 5-8

LOCAL AND GLOBAL IMPACT

•40 publications in ISI journals•commercializable products and 2 technology-know-how •Licensing and patents

Expected National Impact • REDUCED COST OF PRODUCTION• REDUCED DEPENDENCY ON IMPORTS• HIGH VALUE FUNCTIONAL HALAL BIOPEPTIDES

FROM FRESHWATER FISH

Budget

Selected papers Ghassem, M., Arihara, K., Babji, A. S., Said, M. & Ibrahim S. (2011). Purification and

identification of ACE inhibitory peptides from Haruan (China striatus) myofibrillar protein hydrolysate using HPLC–ESI-TOF MS/MS. Food Chem. 129: 1770–1777.

Ghassem, M., Arihara, K. & Babji, A. S. (2012). Isolation, purification and characterisation of angiotensin I-converting enzyme–inhibitory peptides derived from catfish (Clarias batrachus) muscle protein thermolysin hydrolysates. International Journal of Food Science and Technology. doi:10.1111/j.1365-2621.2012.03122.x.

Mahmoodani, F., Ghassem, M., Babji, A. S., Yusop, S. M. & Khosrokhavar, R. (2012). ACE inhibitory activity of pangasius catfish (Pangasius sutchi) skin and bone gelatin hydrolysate. J. Food Sci. Technol. DOI 10.1007/s13197-012-0742-8.

Najafian, L. & Babji, A.S. (2012). A review of fish-derived antioxidant and antimicrobial peptides:Their production, assessment, and applications. Peptides (33): 178–185.

See, S. F., Hong, P. K., Ng, K. L., Wan Aida, W. M. and Babji, A. S. Physicochemical properties of gelatins extracted from skins of different freshwater fish species. International Food Research Journal 17: 809-816 (2010).

Ghassem, M., Arihara, K. & Babji, A.S. 2012. Isolation, purification and characterisation of angiotensin I-converting enzyme-inhibitory peptides derived from catfish (Clarias batrachus) muscle protein thermolysin hydrolysates. International Journal of Food Science & Technology. 47, 2444-2451.

Ghassem, M., Babji, A.S., Said, Mamot. Mahmoodani, F. & Arihara, K. 2013. Angiotensin I-Converting Enzyme Inhibitory Peptides from Snakehead Fosh Sarcoplasmic Protein Hydrolysate. Journal of Food Biochemistry. ISSN 1745-4514.

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