EPSILISEEN® Think natural.® ε-Polylysine

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Siveele B.V.

Think natural.®

ε-Polylysine, the main active ingredient in Epsiliseen®, is a naturally occurring

homo-polymer of L-lysine with a degree of polymerization of 25~35 and

molecular weight of approximately 5000. It is produced by bacterial fermentation

and it has an antimicrobial effect against yeast, fungi and gram-positive and

negative bacteria, making it suitable for versatile applications in the food, feed,

nutraceutical and cosmetic industry.

Description

The way of manufacturing ε-Polylysine through a natural fermentation process was

first described in Japan at the end of 1970s, and since the late 1980s this homo-

polypeptide is approved for use in food by the Japanese Ministry of Health, Labour

and Welfare as a preservative. In 2004, it became GRAS (Generally Recognized As

Safe) certified in the United States of America. As of today, ε-Polylysine is

approved as a food additive in Japan, USA, Korea and some more countries.

History

Chemical specification

IUPAC name

Poly[imino[(2S)-2-amino-1-oxo-1,6-hexanediyl

Properties

Molecular formula (C6H12N2O)n

Molar massvariable 4700 g/mol (degree of polymerization = 30)

Melting point 172.8 °C, 446 K, 343 °F

Basicity (pKb) 5

ε-Polylysine attaches to the cell membrane of the microbial, followed by a destroy

of the complete membrane structure and the option to the material to damage the

energy metabolism which will cause the material, energy and information

transmission of the cell to stop. This will eventually leads to the intracellular

lysosomal membrane rupture, induce the micro-organisms to autolysis and the

break of the Intracellular lysosomal membrane causing death of the bacterial cell.

Function

•ε-Polylysine will be decomposed into lysine and completely digested by human

body.

•Acute oral administration toxicology study in mice shows that ε-Polylysine is

non-toxic.

•According bacterial reverse mutation determination research, ε-Polylysine has no

mutagenicity.

•ADME study shows that the ε-Polylysine has a very poor absorption on the

gastro-intestinal area, and do not accumulate in various tissues and organs.

Safety

Epsiliseen®, a milky powder, has a slight bitter taste. It is soluble in water, slightly

soluble in ethanol, but does not dissolve in ethyl acetate, ethyl ether and other

organic solvents.

Solubility

Epsiliseen® has high thermal stability. The test of antibacterial activity to E.coli

shows very little difference in conditions of 80℃ for 60min, 100℃ for 40min and

120℃ for 40min, as it can hardly be decomposed and inactivated under high

temperature. This allows Epsiliseen® can be treated with the raw material for

sterilization and preventing secondary contaminations.

Stability - Temperature factor

0

2

4

6

8

10

12

14

The effect of temperature to the anti-bacterial activity of Epsiliseen®80℃ (60min) 100℃ (40min) 120℃ (20min) 120℃ (40min)

Inhibition zone diameter

Stability - pH factor

0

2

4

6

8

10

12

14

The effect of pH to the anti-bacterial activity of Epsiliseen®3 4 5 6 7 8 9

Inhibition zone diameter

Some research shows the combination of ε-Polylysine with acidic polysaccharides,

hydrochloric acid salts, phosphate type or copper ions, may causes lower activity

of ε-Polylysine. Protein also has some affect on this. But with hydrochloric acid,

citric acid, malic acid, glycine and senior fatty glyceride, ε-Polylysine becomes

more effective.

Other factors

Compared with Nisin and other natural preservatives, ε-Polylysine has a wide

antibacterial spectrum and there is an obvious killing effort to gram-positive and

gram-negative bacteria, yeasts, molds and viruses. It has a good antibacterial effect

to gram-negative bacteria E. coli and salmonella, which are hard to be controlled

by other natural preservatives. It also has inhibitory effect to the heat resistance of

Bacillus and some viruses, but higher concentration is required to inhibit yeasts

and moulds.

Effectiveness

Minimum Inhibitory Concentrations of ε-Polylysine Against Various Fungi and Bacteria*

Organism mg/kg

Fungi Aspergillus niger IFO4416 250

Trichophyton mentagrophytes IFO7522 60

Candida acutus IFO1912 6

Phaffia rhodozyma IFO10129 12

Pichia anomala IFO0146 10

P. membranaefaciens IFO0577 3

Rhodotorula lactase IFO1423 25

Sporobolomyces roseus IFO1037 3

Saccharomyces cerevisiae 50

Zygosaccharomyces rouxii IFO1130 150

Gram Positive Bacteria Geobacillus stearothermophilus IFO12550 5

Bacillus coagulans IFO12583 10

B. subtilis IAM1069 3

Clostridium acetobutylicum IFO13948 32

Leuconostoc mesenteroides IFO3832 50

Lactobacillus brevis IFO3960 10

L. plantarum IFO12519 5

Micrococcus luteus IFO12708 16

Staphylococcus aureus IFO13276 12

Streptococcus lactis IFO12546 100

Gram Negative Bacteria Raoultella planticola IFO3317 8

Campylobacter jejuni 100

Escherichia coli IFO13500 50

Pseudomonas aeruginosa IFO3923 3

Salmonella typhimurium 16

*Source: Yoshida T., Nagasawa T. (2003). ε-Poly-L-lysine: microbial production, biodegradation and application potential. Appl.

Microbiol. Biotechnol. 62(1):21-26.

Epsiliseen®-D

Epsiliseen®-D is a natural antimicrobial compound of ε-Polylysine and dextrose, used in many food

applications to effectively inhibit moulds, yeasts, Grampositive and Gram-negative bacteria.

Epsiliseen®-H

Epsiliseen®-H is a natural antimicrobial agent with ultra-high concentration of ε-Polylysine, used in

food, cosmeceutical and pharmaceutical applications.

EPSILISEEN® Think natural.® ε-Polylysine

•Natural and non-animal source

•Broad-gauge antibacterial spectrum, especially Gram - negative bacteria

•Extends shelf life of food

•Effective over a wide range of pH values (4-10)

•Can be used before or after heat treatment due to excellent temperature stability

•Secures desired food flavor

•Improves cost efficiency due to the low dosage rate used

•Meets consumer demands for food protected with natural additives by fully or

partially replacing synthetic additives

Key benefits in food applications

Food Applications of Epsiliseen® Table 1

Applications Use-level (%)*Alcoholic beverages Light beer 0,01

Wine 0,005

Baked goods and baking mixes Cake batter and cheesecakes 0,01

Muffins and popovers 0,01

Pancakes and waffles 0,025

Pies (not fruit) 0,025

Sweet pastries 0.01 - 0.025

Beverages and beverage bases Energy, fitness, sports, and isotonic drinks (ready to drink) 0,025

Gelatin, rice, and sugar cane-based beverages 0,025

Soft drinks 0,025

Cheeses Cheese-based mixtures 0,025

Cottage, ricotta and cream cheese 0,025

Natural (Feta, Mozzarella, and Mexican) 0,025

Processed cheese and cheese spreads 0,025

Coffee and tea Coffee and coffee beverages 0,025

Tea and tea beverages 0,025

Condiments and relishes Pickles, olives, and pickled products 0,025

Dairy product analogs Cream substitutes 0,025

Imitation cheese 0,025

Egg products Egg-based dishes 0,025

Fats and oils Reduced-fat or fat-free margarine-like spread 0,025

Salad dressings (regular and low-fat) 0,025

Fish products Fish and shellfish-based foods 0,025

Fresh eggs Eggs 0,025

Fresh fish Fish and shellfish 0,025

Fresh meats Beef (steak, stewing, and ground) 0,06

Pork (chops and steaks) 0,06

Fresh poultry Chicken breast and turkey 0,06

Fruit and water ices Frozen fruit bars and sorbet 0,025

Food Applications of Epsiliseen® Table 2

Applications Use-level (%)*Gelatins, puddings, and fillings Gelatin 0,025

Pie fillings 0,025

puddings, custards, and mousses 0,025

Grain products and pastas Grain-based dishes 0,025

Pastas 0,025

Gravies and sauces Fat-based sauces 0,01

Gravy 0,025

Tomato-based sauces 0,025

Meat products Meat-based prepared foods 0.02 - 0.03

Milk and milk products Chocolate-flavored drinks 0,01

Cream 0,01

Eggnog 0,01

Sour cream and dips 0,025

Yogurt 0,01

Nuts and nut products Peanut butters and sauces 0,025

Plant protein products Meat substitutes 0,025

Soy products 0,025

Poultry products Poultry-based prepared foods 0,025

Processed fruits and fruit juices Fruit-flavored drinks and Ades (ready to drink) 0,025

Fruit smoothies 0,025

Nectars 0,025

Prepared fruit and fruit mixtures 0,025

Processed vegetables and vegetable juices Dry beans and bean-based prepared foods 0,025

Potato and sweet potato-based foods (excluding French fries) 0,025

Vegetable-based prepared foods 0.01 - 0.025

Tomato-based juices 0,025

Soups and soup mixes Soups (excluding canned) 0,025

Sweet sauces, toppings and syrups Sweet syrups and toppings 0,025

* Use levels were based on 200 mg EpsiliseenTM-D / RACC; RACC=Reference Amounts Customarily Consumed per Eating Occasion (21 CFR § 101.12 - U.S. FDA, 2009b). When a range of values is reported for a food application, particular foods within that application may differ with respect to their RACC.

ε-Polylysine was assigned INCI name as POLYEPSILON-LYSINE by the

International Nomenclature Committee in 2009, and classified as hair

conditioning and skin conditioning.

Personal care applications of Epsiliseen®

•Natural and safe

•Effectively inhibits Gram - positive, Gram - negative bacteria, yeasts and moulds,

especially Gram - negative bacteria

•Easy to use

•Meets consumer demands for natural products with natural additives by fully or

partially replacing synthetic additives

Key benefits in personal care applications

Personal Care Applications of Epsiliseen®

Applications Use-level (%)*

Skin care (emulsions) 0,05

Hair care (emulsions) 0,05

Lotions 0,05

Organic products 0,05

Siveele B.V.

Minervum 7113, 4817 ZN Breda, The Netherlands

t: +31 (0) 765710222

f: +31 (0) 765713788

www.siveele.com info@siveele.com

For further information, please contact Siveele B.V. or our local representative.

Non Warranty: The information contained herein is provided in good faith and, to the best of our knowledge, is true and correct. It may be subject to change without further notice. This information is offered solely for your consideration and verification. Siveele B.V.