levan moist eng ts
TRANSCRIPT
1. What is Levan ?
Levan is - Biopolymer existing in natural plants or produced by micro-organisms
- Several thousand to hundred thousand fructose linkages
- Excellent properties as a water soluble dietary fiber
Plant sourced Levan - fermented soybeans
- cereals (barley and wheat)
- vegetables (onion, chicory, garlic)
Micoorganisms sourced levan Zymomonas mobilis / Bacillus subtilis
/ Rahnella aquatilis / Pseudomonas syringae
Sucrose
levansucrase
LEVANFru
Fru
DFA IV
FruGlu Fru Fru Fru Fru Fru Fru Fru Fru Fru
FruFru
Fru Fru Fru Fru Fru Fru
Fru Fru Fru FruGlu
o
H
HO H
O H
C H 2
C H 2O H
Oo
H
HO H
O H
C H 2
C H 2O H
O
H
C H 2
o
H
HO H
O H
C H 2O
H
H
HHH
o
H
HO H
O H
C H 2
C H 2O H C H 2
O
O
H
HO H
O H
o
O
H
H
C H 2
O H
O HC H 2O H
o
H
O
H
H
C H 2
O H
O HC H 2O H
o
H C H 2O H
β-(2,6) fructose polymer containing β-(2,1) branches
2. Application
PharmaceuticalsFunctional Food
Animal Feed Industrial
▪ Dietary ▪ Prebiotic ▪ Medical Food (for diabete)
▪ Natural adhesive▪ Detrgent and shampoo▪ Reforming agent for plas-tics
▪ Pig▪ Poultry, Cow, Pet dog ▪ Prebiotic, immune, biomineral
Cosmetics
▪ NMF as HA, B-Glucan▪ Skin trouble, Anti-inflammatory ▪ Cell proliferation
▪ DDS expients▪ Enhencing bio minerals▪ Bio adhesive (Hydrogel)
3. Levan moisture for skin care
▪ Levan is a sugar polymer with β-(2,6) fructose polymer that produced by Levansucrase from sugar. - world first patent : Method for producing levan using levansucrase (Pat No. 1001459460000 /1998) - Patent - Cosmetic compositions containing levan ; Korean pat. 1004403570000 - Key technology : microorganism, gene cloning, levansucrase, purifying, mass production
▪ Product types : Levan HM (high molecular & Milky type) Levan LM(Clear type)① ②
▪ Effect : Natural moisture, Cell-Proliferation, Anti-Inflammation, Non-Toxic
▪ CTFA/INCI name : Levan (CTFA monograph ID 19108) CAS No. 9013-95-0 ▪ Application : skin care(lotion, essence, cream), facial mask, lip care, mascara
Impaired barrier
Elevated TEWL
Cytokine release
Eczema
Dermal inflammation
Cytokine influence
Immediate repair
Moisture Barrier
Delayed repair
1 차 Blocking
2 차 decreasing
Levan Skin MechanismSkin Mechanism
4. Clinical Effects
1) 보습효과1) 보습효과
1. Moisturizing effect measured by weight change method + Put 5 grams of each sample in a desiccator filled with
CaCl2 and leave them under R.H 65%, 37 and measure℃ weight changes.
2. Skin moisture content by using Corneometer CM825 + Temperature 20 , Relative humidity 20%℃ + Measure the change of moisture content of skin (forearm) at regular intervals
2) Cytotoxicity & Proliferation2) Cytotoxicity & Proliferation
4. Clinical Effects
A. Cytotoxicity of Levan by MTT assay on human skin fibroblast
B. Promotion of Keratinocyte differentiation by Levan
▪ The figure shows proliferation of fibroblast by Levan.
▪ It means Levan is non-toxic.
▪ Human Keratinocyte Haca-T cell → MTT assay ► Fructan promotes differentiation of keratinocyte cell.
3) Cell proliferation test with 3-Dimentional artificial skin3) Cell proliferation test with 3-Dimentional artificial skin
A : Control
B : 0.05% SLS
C : 0.05% SLS + 0.01mg/ml Levan
D : 0.05% SLS + 0.05mg/ml
Levan
(MTT conversion assay)
A B C D75
80
85
90
95
100
▪ We determined that Levan treated artificial skin exhibited greater cell proliferation.
(more than a 30% enhancement)
4. Clinical Effects
4) Anti-Inflammation test with 3-Dimensional artificial skin 4) Anti-Inflammation test with 3-Dimensional artificial skin
A : Control
B : 0.05% SLS
C : 0.05% + 0.01mg/ml Levan
D : 0.05% + 0.05mg/ml Levan
A B C D0
10
20
30
40
50
60
70
4. Clinical Effects
▮ After inducing primary skin irritation with each sample in 3-D artificial skin and we subsequently measured the quantity of interleukin-1α secreted.
▪ Our results an emollient effect during skin irritation caused by irritants. → anti-inflammatory effect
5. Safety test
Human skin Primary Irritation Test Human skin Primary Irritation Test
Tested materialNo. of
responder
48hr 72hr 반응도
1+ 2+ 3+ 1+ 2+ 3+ 48H 72H Mean
Levan 0 - - - - - - 0.0 0.0 0.0
Results of human skin primary irritation test
▣ During clinic test for skin safety evaluation
Result : Test product is classified as non-irritating range for human skin primary irritation. Thus, these products can be considered to be safe.
Symptom 4weeks 8weeks
Subjective irritationItching, Prickling, Burning, Stinging, StiffnessThightening, Burning of eyes, Weeping, etc All “0” All “0”
Objective irritation Erythema, Ederma, Scale, Papule, etc. All “0” All “0”
The number of subjects 0 0
6. Stability test of Levan in Ethanol
10% 20% 30% 40% 50%
4℃(Lower)
10% 20% 30% 40% 50%
25℃(room temp)
10% 20% 30% 40% 50%
40℃(Higher)
7D 15D 30D 45D
10% [ stable ]
20% [ stable ]
30% [ stable ]
40% [ stable ]
50% [ stable ]
days
Ethanol %
< 표 1 >
• Levan is a hydrophilic polysaccharide and stable in ethanol
without any change in the Levan characteristics.
7. Particle size of Levan solution
Weight Number
1) 5% Levan : Diameter 224.3nm1) 5% Levan : Diameter 224.3nm
2) 5% Levan in 20wt% aqueous ethanol : 251.8nm 2) 5% Levan in 20wt% aqueous ethanol : 251.8nm
Weight Number
TEM picture of nanoparticles
8. Patent List (Intellectual property)
No Title of invention Registration No. Status
1 Method for producing levan using levansucrase 1001459460000 Registered
2 Novel polyethylene glycol/levan aqueous two-phase system and protein partitioning method using thereof 100262690000 Registered
3-1 Levan fructotransferase derived from Arthrobacter ureafaciense and process for preparing difurctosedianhydride IV from levan by using same
1003809700000 Registered
3-2 Enzymatic production of difructose dianhydride IV form sucrose and relevan enzymes and genes coding for them (US)06841368 Registered
3-3 Enzymatic production of difructose dianhydride IV form sucrose and relevan enzymes and genes coding for them (WO)2001029185 Pending
3-4 Enzymatic production of difructose dianhydride IV form sucrose and relevan enzymes and genes coding for them (CN)1192100 Registered
3-5 Enzymatic production of difructose dianhydride IV form sucrose and relevan enzymes and genes coding for them (EP)01141236 Pending
4 Process for the preparation of low-molecular weight levan by using immobilized levansucrase or microorganismcells treated with an organic solvent
1003238370000 Registered
5-1 Animal feed containing simple polysaccharides 1003426550000 Registered
5-2 Animal feed containing simple polysaccharides (WO)2001049127 Pending
5-3 Animal feed containing simple polysaccharides (US)20030086959 Pending
6 Cosmetic compositions containing levan having cell-proliferation, skin-moisturizing and irritation-alleviating effect 1004403570000 Registered
7 Process for purification of levan using ultra membrane system 1004348840000 Registered
8 A process for the extra-cellular secretory production of Zymomonas mobilis levansucrase 1004964190000 Registered
9 Dietary composition including levan for prevention and treatment of obesity 1005001580000 Registered
10 Process of levan production using novel levansucrase 1004203130000 Registered
11 Functional levan chungkukjang powder and the manufacturing method 1020090033937 Pending
12 Rice cooking additive containing Levan and DFA IV 1020100094078 Pending
Ref. 2. Application