Cyclodextrins in biotechnologyLankacidin A (R=Ac) Lankacidin C (R=H) Lankacidinol A (R=Ac)...
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Cyclodextrins in biotechnology
Transcript of Cyclodextrins in biotechnologyLankacidin A (R=Ac) Lankacidin C (R=H) Lankacidinol A (R=Ac)...
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Cyclodextrins inbiotechnology
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What are cyclodextrins (CDs)?
• Composed of sugar units• Cyclic, doughnut-shaped molecules• Naturally occurring compounds produced from plants (no animal origin!)• Used in food, pharmaceuticals, drug delivery, chemical industries,
agriculture, etc.
Reversible inclusion complex formation
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Why use cyclodextrins?
- CDs enhance the solubility of complexed substrates (substitute detergents and co-solvents)
- CDs do not damage the microbial cells or the enzymes
- CDs intensify the enzymatic conversion of lipophilic substrates
- CDs improve the yield of product-inhibited fermentations
- Organic toxic compounds are tolerated by microbes in higher concentrations
- Compounds in small amounts can be isolated simply from complicated mixtures
- CDs complexes can substitute for mammalian serum in tissue cultures
- Unstable and/or insoluble proteins can be dissolved and stabilized in aqueous solution
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Who we are and what we offer?
CycloLab is the world's only all-around Cyclodextrin Service Provider
Our services include:• Supplying cyclodextrins for commercial products and product
development;• Screening cyclodextrin derivatives to find the right candidate for
the request of customers;• Providing formulation development services, composition
optimization, stability assessment;• Offering analytical services to characterize complexes and
products;• Preparing pilot-scale amounts for cyclodextrin complexes under
GMP for development purposes;• Assisting in compilation of regulatory documentation.
For more information please click here
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Microbiological transformation
• Enhanced
conversion rate
• Decreased product
inhibition
• Improved product
stability
60
80
100
0.5 1 1.5 2 2.5 3 3.5
20
40
Molar ratio β-cyclodextrin/cholesterol
AD
(%
mola
r yie
ld)
120 h
180 h
60 h
Bar: Trends Biotechnol.
1989, 7, 24
Cholesterol
Feedback inhibition
Micobacterium sp.
Side chains degradation
4-Androstene-3-17-dione1,4-Androstadiene-3-17-dione
+
New steroid drugs
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Biosynthesis by fermentation
Streptomyces rochei volubilis
Lankacidin A (R=Ac)Lankacidin C (R=H)
Lankacidinol A (R=Ac)Lankacidinol C (R=H)
Macrolides Antitumor-Antibiotic
0
0,5
1
1,5
2
2,5
3
0 0,5 1 1,5 2 CD (%)
Lankacid
inpro
duction (
mg/m
L)
βCD
γCD
αCD
control • Enhanced
production of
antibiotics
• Diminished by-
products formation
• Decreased product
inhibition
Sawada et al.: Appl.
Microbiol. Biotechnol.
1990, 32, 556559
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Vaccine production
Bordetella pertussisPertussis toxininhibition
Production
Palmitic acid
Oleic acid
Inoculumsize cells in
5 μL
0 α β γ DIMEB
103
104
105
106
107
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- no growth + < 100 colonies ++ 102 to 103 colonies +++ full growth
Bordetella pertussis cell growth
DIMEB (dimethyl beta-cyclodextrin) increases pertussin toxin production 100-fold!Imaizumi et al.: Infect. Immun. 1983, 41, 1138-1143
Complexation of
fatty acids
(growth
inhibitors)
results in
enhanced cell
growth and
toxin production
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Serum-free culture media
• Solubilization of lipids
(fatty acids, cholesterol,
phospholipids)
•No threat of prion proteins
Mycobacterium leprae
Water-soluble lipid/CD complexes:Cultivation of non-cultivable Mycobacterium leprae;
Serum substitutes for lymphoblast cells;
Non-cholesterol interacting fatty acid/CD complexes
Lymphoblast cellsSzente et al.: J. Incl. Phenom. Mol.
Recogn. Chem. 1993, 16, 339-354
Rajnavölgyi et al. Beilstein J.
Org. Chem. 2014, 10, 3152–3160
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Cyclodextrin in artificial fertilization
• Improvement of the quality of semen by cholesterol supplementation with
cholesterol loaded methyl BCD (cryopreservation)
• Enhancement of capacitation and fertility rate by preincubation of thawed sperms
with methyl BCD
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Reducing enzymatic browning of fruit juices
OH
OH
O
O
OH O
OHOH
O
OH
OH
O
chlorogenic acid (and other polyphenols)
Polyphenol oxidase
Browning
• Complete inhibition of
browning by soluble CDs
Hicks et al.: J. Agric. Food Chem.
1996, 44, 2098-2101
Lopez-Nicholas et al.: J. Agric.
Food Chem. 2007, 55, 5312-5319
αCD > maltosyl βCD > βCD
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CD-assisted purification of waste water and soil
• Improved solubility/
bioavailability of the pollutants
• Improved cell growth
• Protection of the biological
sludge in the waste water plants
• Enhanced microbial
decomposition of contaminants
0
20
40
60
80
100
120
0 0.1 0.3 0.5 0.7Tra
nsf
orm
er
oil
co
nc.
in
th
e
soil
re
late
d t
o t
he
sta
rt
RAMEB (%)
Biodegradation of transformer oil in loamy soil
start 3 days 14 days 28 days
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10
20
30
40
50
60
0 20 40 60 80
Bip
henyl conc. (m
g/L)
Time (h)
Biphenyl biodegradation
0
10
20
30
40
50
60
0 20 40 60 80
Bip
henyl conc. (m
g/L)
Time (h)
Biphenyl biodegradation
Yoshii et al.
Biol. J. Armenia,
2001, 18, 226-236
Fenyvesi et al.
Land Contam. Reclam.
2009, 17, 585-597
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Company contacts:
CycloLab Cyclodextrin Research & Development Laboratory Ltd.
Budapest, P.O. Box 435, H-1525 Hungary
Location: Illatos út 7., Budapest, H-1097 Hungary
TEL: (+36) 1-347-60-70; FAX: (+36) 1-347-60-68
E-mail: [email protected]; Homepage: http://cyclolab.hu/
Contact person: Tamas Sohajda
R&D Director
Tel: (+36) 1-347-60-72
