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Enzymatic Hydrolyzed Brown Seaweeds
1. Sulfated Polysaccharides from Brown Seaweed2. Small Molecular Fucoidan
2014-10-05
Mr. Chia-Chin Sheu,President, Simpson Biotech Co., Ltd.
For Internal Evaluation Only, All Right Reserved
Simpson BiotechA Bio-Fermentation Company
For Internal Evaluation Only, All Right Reserved
An R&D oriented and innovative bio-fermentation company established in Dec. 1998 68 years of fermentation experiences (derived from Kong-Yen Vinegar Group)
Employees: 34 Capital: 10 millions US Dollars
Major Businesses Areas:Medicinal fungus/mushroom fermentation and their applications.Purification, identification, and the applications of the active ingredients isolated from the fermented medicinal fungus mycelia.Bio-transformation & Bio-conversion to produce functional ingredientsUse HERPEPSTM to produce recombinant proteins including industrial enzymes, pharmaceutical proteins and biosimilarsOEM/ODM services, i.e., producing tailor-made recombinant proteins/enzymes by using HERPEPSTM platform
About Simpson Biotech
For Internal Evaluation Only, All Right Reserved
All Rights Reserved 4
SimpsonPure Bioactive Natural Ingredients Part 1: Fermented mycelia of Antrodia cinnamomea (A. camphorata)
Part 2: Fermented Cordyceps sinensis Mycelia & Extracts
Part 3. The Ginsenoside Compound K & the Fermented Ginseng
Part 4:
• Water Soluble/Low Molecular Chitosan (COS-LM) – Anti-microbial Agent
• Oligo Chitosan (COS-600) - the Chitobiose & Chitotriose
Part 5. Low-bitterness & Low Mw Peptides:
• Fish Scale Collagen Peptides
• Soy Peptides and Low Salt Seasoning Solutions
• Silk Peptides & others
Part 6. Enzymatic Hydrolyzed Seaweeds: Brown, Red & Green Algae
• Sulfated Polysaccharides from Brown Seaweed – Dietary Supplement & Drinks
• Small Molecular FucoidanFor Internal Evaluation Only, All Right Reserved
Part 6The Enzymatic Hydrolysation of
Seaweed into Low Molecular Functional Polysaccharides& Other Active Ingredients
- The Sea Weed Hydrolyzation Platform Technology
For Internal Evaluation Only, All Right Reserved
Seaweed polysaccharide hydrolyzation technology – for Brown, red & green seaweeds
Red Seaweed:
Brown Seaweed:
Green Seaweed:
www.topnews.in/usa/algae-knew-about-quantum-m...
www.liveaquaria.com/product/prod_display.cfm?..
oceanexplorer.noaa.gov/.../media/sargassum.html
For Internal Evaluation Only, All Right Reserved
What is Fucoidan?
1. Fucoidan is a sulfated polysaccharide (MW: average 20,000) found mainly in various species of brown seaweed such as mozuku, kombu, limu moui, bladderwrack, wakame, and hijiki
2. contains substantial percentages of L-fucose and sulfate ester groups, mainly derived from brown seaweed
3. Fucoidan is just a general term for the polysaccharide composed of fucose and sulfate ester plus galactose, annose, xylose, uronic acids, etc. 其化學成份為以硫酸岩藻糖為主的多醣體,結合岩藻糖以外的Galactose、Annose、Xylose、Uronic Acids等。同一構造的物質不能全稱做Fucoidan,它是岩藻糖鏈的總稱,至今還無法定出Fucoidan褐藻糖膠的定義。(Robertet al., 2004)。
Major interesting: antitumor and immunomodulatory
For Internal Evaluation Only, All Right Reserved
Structure of Fucoidan
褐藻醣膠 fucoidan oligosaccharide
For Internal Evaluation Only, All Right Reserved
資料來源:Mar. Drugs 2011, 9, 196-223
Fucoidan oligosaccharide
For Internal Evaluation Only, All Right Reserved
Biological activities of Sulfanted Polysacchrides from Brown Seaweed
Diabetes Control: α-Amylase & α-Glucosidase inhibitor for Type 2 diabetes. Anti-Anticoagulant and antithromboticAnti-virusAnti-tumor and immunomodulatoryAnti-inflammatoryBlood lipids reducingAntioxidant activitiy against hepatopathy, uropathy and renalpathyGastric protective effects
For Internal Evaluation Only, All Right Reserved
Previous Research on Seaweed & Seaweed Polysacchrides -1
Anticoagulant effects:Carbohydr. Res. 2007, 342, 2326-2330.Glycobiology 2005, 15, 369-381.
Anti-viral activity:Carbohydr. Res. 2007, 342, 2326-2330.Glycobiology 2005, 15, 369-381.
Anti- Tumor activity Am. J. Hematol. 2004, 78, 7-14.Bull. Exp. Biol. Med. 2007, 143, 730-732.Anticancer Res. 2005, 25, 2129-2133.
Immuno-regulation:J. Health Sci. 2005, 51, 394-397.Immunol. Lett. 2008, 115, 138-143.J. Med. Food 2005, 8, 446-453.
For Internal Evaluation Only, All Right Reserved
Anti-oxidation:Chin. Trad. Herbal Drugs 2003, 34, 824-826.J. Appl. Phycol. 2007, 19, 153-160.High Tech. Lett. 2005, 11, 91-94.Int. J. Biol. Macromol. 2008, 42, 127-132.
Hypolipidemic effect:Food Sci. 1999, 20, 45-46.Food Sci. 2001, 22, 92-95.J. Ocean Univ. Qingdao 2004, 34, 560-564.
Anti-inflammation activity:Int. Arch. Allergy Immunol. 2005, 137, 289-294.Biochem. Biophys. Res. 2006, 346, 345-350.
Liver-protective activity:Hepatol. Res. 2006, 35(3), 190-198.Pharmacol. 2008, 580, 380-384.
Previous Research on Seaweed & Seaweed Polysacchrides -2
For Internal Evaluation Only, All Right Reserved
Simpson Biotech’s Enzymatic Hydrolysation Process
Brown Seaweed Material wash Grinding add water & food grade enzymes mixing & reaction Sterilization reduce to room temperature press/filtration Hydrolzyted Clear Broth Pasteurization Frozen StorageBS Hydrolyzed Conc. BSH Conc.
Hydrolyzed Clear Broth Concentration add 4x Alcohol to get precipitation (BS Polysacchrides Powder, BSPS) & Clear broth without polysacchrides (Non-Poly Broth) Non-Poly Concentrated to get Non-Poly Conc. Liquid, BSNoPS Conc. Pasteurization Frozen Storage.
15
Molecular Wt. Distribution (HPLC/GPC, RI detector) of Simpson Biotech Enzymatic Hydrolyzed Brown Seaweed (EHBS) and Commercial products善笙酵素水解褐藻多醣與市售產品之分子量比較圖
0.5Kd
善笙酵素水解褐藻多醣兼具有市售高分子多醣(日本T廠800~1200Kd)及小分子寡糖(台灣H廠0.5Kd)之特性,且獨具有特殊中分子多醣(10~50Kd) ,為其他廠牌商品所沒有。
Simpson EHBSJapan T BrandTaiwan H BrandAustralia M BrandJapan S Brand
0.5Kd
16
Immunoregulatory Activities (Antibody IgM stimulation) of Polysaccharides from Simpson Biotech’s Enzymatic
Hydrolyzed Brown Seaweed (PS-EHBS)
Fig 1. PS-EHBS effectively induce the antibody (IgM) secretion in a human hybridoma cells HB4C5 assay
0
200
400
600
800
1000
1200
*
******
Concentration (micro g/mL)
IgM
con
cent
ratio
n(ng
/ml)
17
Immunoregulatory Activities (Anti-inflammation) of Polysaccharides from Simpson Biotech’s Enzymatic
Hydrolyzed Brown Seaweed (PS-EHBS)
ClaAPS
Contr
ol 0 125 250 5000
10000
20000
30000
40000
LPS (1 g/mL)
#
******
Concentration (μ g/mL)
TNF-
α c
once
ntra
tion(
pg/m
l)
ClaAPS
Contr
ol 0 125 250 500
0
5000
10000
15000
20000
LPS (1 g/mL)
#
*****
Concentration (μ g/mL)
IL-6
con
cent
ratio
n(pg
/ml)
Fig 2. PS-EHBS effectively reduce the LPS-induced Superoxide in a murine RAW 264.7 macrophage cell line assay
Fig 3. PS-EHBS effectively inhibit both the cytokines TNF-α and IL-6 in a LPS-stimulated RAW264.7 macrophage cell line assay
0.0
0.2
0.4
0.6
LPS (1g/mL)
#
******
******
**
Concentration (?g/mL)
Supe
roxi
de (O
D. 6
30 n
m)
18
-
10.0
20.0
30.0
40.0
50.0
60.0
70.0
ControlPS-EHBS-A61.7mg/Kg
NK
cyt
olys
is, %
.
E/T=25E/T=50E/T=100
******
************
****
*** * *
PS-EHBS-A205.6mg/Kg
PS-EHBS-F205.6mg/Kg
EHBS-A205.6mg/Kg
Immunoregulatory activity (Natural Killer) of 2 different (A & F) Enzymatic Hydrolyzed Brown Seaweeds (EHBS)
and its Polysaccharides (PS-EHBS) on Miceat the Dosages Levels Equal to 0.3 and 1.0g/day/person
19
0.0%
10.0%
20.0%
30.0%
40.0%
50.0%
60.0%
70.0%
80.0%
90.0%
100.0%
Control
Pha
gocy
tosi
s, %
.
MOI=0MOI=1MOI=5MOI=10
*
*
** ** **
Multiplicity of infection
PS-EHBS-A61.7mg/Kg
PS-EHBS-A205.6mg/Kg
PS-EHBS-F205.6mg/Kg
EHBS-A205.6mg/Kg
Immunoregulatory activity (Mononuclear Phagocytic) of 2 different (A & F) Enzymatic Hydrolyzed Brown Seaweeds
(EHBS) and its Polysaccharides (PS-EHBS) on Miceat the Dosages Levels Equal to 0.3 and 1.0g/day/person
α-Amylase & α-Glucosidase inhibition Activity of Simpson Biotech’s Enzymatic Hydrolyzed Brown
Seaweeds Polysaccharide & Commercial Products
Enzymes Inhibition Activities
α-Amylase Inhibition (IC50)
α-Glucosidase Inhibition (IC50)
Cebolixs by InSea2 52.8μg/ml 75.0μg/mlSimpson Hydrolyzed Brown Seaweed #1 27.4μg/ml 156μg/ml
Simpson Hydrolyzed Brown Seaweed #2 6.36μg/ml 83.9μg/ml
Commerical Fucoidan Ingrident #1 no activity
few activity (22% inhibition in 5mg/mL)
Commerical Fucoidan Ingrident #2 no activity no activity
Commerical Fucoidan Product #3 no activity no activity
For Internal Evaluation Only, All Right Reserved
21
Starch Management: (α-Amylase inhibition)Clinical Trials for 23 healthy participants1. Reduce 48% blood glucose2. Reduce 12% insulin secretion3. Increase 8% Cederholm index (insulin sensitivity)
Competing Commercial Product - InSea2®- Active ingredients isolated from similar brown seaweeds
Sugar Management: (α-glucosidase inhibition)Clinical Trials for 42 healthy participants)1. Reduce 39% blood glucose2. Reduce 7% insulin secretion3. Increase 5% Cederholm index (insulin sensitivity)
Weight Management: Clinical Trials for 39 healthy (BMI 24~40) participants1. Increase 33% fullness2. Reduce 14% ingested calories3. Significant weight loss4. Safety: well tolerated for 3 month at daily
dosage = 1000 mg
The extraction yield (EY) & crude polysaccharides(CPP) molecular weight distribution (MWD) of all kind of
seaweeds (1.Green algae 海葡萄 )
For Internal Evaluation Only, All Right Reserved
The extraction yield (EY) & crude polysaccharides’ molecular weight distribution (MWD) of all kind of seaweeds
(2. Red algae 紫菜 )
For Internal Evaluation Only, All Right Reserved
The extraction yield (EY) & crude polysaccharides’ molecular weight distribution (MWD) of all kind of seaweeds
(3. Brown algae 海带 )
For Internal Evaluation Only, All Right Reserved
The extraction yield (EY) & crude polysaccharides’ molecular weight distribution (MWD) of all kind of seaweeds
(4. Brown algae 马尾藻 )
For Internal Evaluation Only, All Right Reserved
The extraction yield (EY) & crude polysaccharides’ molecular weight distribution (MWD) of all kind of seaweeds
(5. Brown algae 裙带菜 )
For Internal Evaluation Only, All Right Reserved
The extraction yield (EY) & crude polysaccharides’ molecular weight distribution (MWD) of all kind of seaweeds
(6. Brown algae 昆布 )
For Internal Evaluation Only, All Right Reserved
Thanks …Q&A ?
Contact to: Mr. Chia-Chin Sheu (許嘉欽)Simpson Biotech Co., Ltd. 善笙生物科技股份有限公司
[email protected]:+886-3-396-3466
For Internal Evaluation Only, All Right Reserved