SABMiller has grown into a global
company with operations and
distribution agreements across six
continents. It is one of the world’s
top 3 largest brewers, and its
portfolio of brands Pilsner Urquell,
Peroni Nastro Azzurro, Castle, Miller
Genuine Draft and Grolsch.
Local beverage market is saturated,
and therefore are interested in some
diversification into biotech,
especially around byproducts.
Tongaat Hulett Starch is Africa’s largest producer of Starch,
Glucose and related products. It uses enzymes for
production of glucose and fructose syrup.
Hulett Sugar has a central refinery in South Africa with an
annual refining capacity of some 600 000 tons with
refining capacity at other sites. Also provides industrial
sweeteners Acesulfame K, Aspartame, Sodium Cyclamate
Sodium Saccharin, Polydextrose
Illovo: Sugar and by products: Furfural, Furfuryl Alcohol,
Diacetyl, 2,3 – Pentanedione, Methanol, Ethyl alcohol
(Ethanol).
TSB: Sugar
SMRI: Joint research institute, seeking new biotechnology
options, particularly natural polymers.
AECI (SA Bioproducts)
11 000 tonne/year lysine
plant at Umbogintwini,
Natal province, South
Africa.
Ciba-BASF nitrile hydratase
based biocatalysis technology
has been implemented by
SENMIN in South Africa.
Senmin’s new plant, Sasolburg, generates acrylamide from acrylonitrile
UK and SA. BBI Enzymes is the market
leader in the manufacture of natural
enzymes to a global marketplace.
Natural Enzymes
SA SME start up
Importer
Fermentation for
Ethanol
Spirit vinegar
Fusel oils
Interested in Oxidation of alkanes or alkenes and Carbon dioxide use.
Environmental biotechnology.
Chem City – research and SME incubator.
Fragrances, flavours and oils.
Products and research include
application of biocatalysis
(Steenkamp et al (patent)).
Vaccines under development include a liquid
pentavalent vaccine (diphtheria, tetanus,
pertussis, hepatitis B and Haemophilus
influenza type b.
EPO, Monoclonal Antibodies
Streptomyces antibiotics:
Actinomycin D ; Brefeldin A ; Ionophores;
Leptomycin B etc.
• Bioprospecting: Natural Products Chemistry
• Emerging Health Technologies (including
synthetic biology)
• Protein Technologies:
– Vaccines
– Antibodies
– Enzymes
– Biocatalysis
• Sunlight and dry areas
useful for open ponds.
• Βeta-carotene production
using algal technology.
Natural Carotenoids South
Africa (Pty) Ltd small plant
built in Uppington.
•
•
Commercial scale
Custom production of plant made proteins
• For a developing country, South Africa has an unusually large chemical
industry and it is of substantial economic significance. In 2010 the
industry still contributes 5% to the GDP and 25% of manufacturing sales.
• The chemical industry is focusing on the need to be internationally
competitive and the industry is reshaping itself accordingly. Exports were
R49 billion and imports R95 billion in 2009 with the gap declining.
• Local Giants SASOL, AECI, Sentrachem (DOW), OMNIA (Protea) (and
about 1000 small companies).
• Many multi-national companies operate in South Africa as manufacturers
and/or distributors. Amongst these companies are Air Products, Bayer,
BASF, Buckman, du Pont, Huntsman Tioxide, Lanxess, Merck and Rohm
and Haas (now part of Dow).
(Source CAIA 2010)
• Environmentally Friendly Technology
• Aim to enhance the fine chemicals and
Pharmaceutical sectors.
• Interest in renewables
Hydrogenation of thymol
produces an 8 isomer mixture of
the enantiomers of menthol,
isomenthol, neomenthol and
neoisomenthol).
CSIR patented process utilises
an enzyme in organic media to
enantioselectively and
diastereoselectively catalyse
the separation of (–)-menthol
from the post hydrogenation
mixture
© CSIR 2011
www.csir.co.za
OH
O
N
O
O
N
2 step 2 enzyme 1 pot
high yielding
biocatalytic reaction
in a slurry.
NH
+
N
H
NH
N
OH
HO
NH2
Guanosine
O
Thymine
Purine
nucleoside
phosphorylase
PO43-
Uridine
nucleoside
phosphorylase
20% solid loading
~90% yield
O
O
NH
N
NH
+
OH
N
O
O
OH
HO
5-Methyluridine
N
H
N
N H2
Guanine
AcBr CH3CN,
reflux 1 h
O
O
NH
OAc
O
N
O
DABT
© CSIR 2011
www.csir.co.za
MeOH, H2 9 bar,
Ni catalyst 1-2 h
O
NH
OAc
O
Br
OAc
Integrated into
synthesis of
thymidine, stavudine
and AZT
N
O
NH
OH
O
N
MeOH, NaOMe,
40°C, 1 h
OAc
HO
DAT
Thymidine
O
PNP from
halophile
Directed
evolution
of UP from
E. coli
Visser DF, Hennessy F, Rashamuse K, Louw M and Brady D. (2010) Cloning,
purification and characterisation of the purine nucleoside phosphorylase
from Bacillus halodurans Alk36. Extremophiles 14: 185-192
Visser DF, Rashamuse JK, Hennessy F, Gordon G, van Zyl PJ, Mathiba K, Bode
ML and D Brady (2010). Cascade Enzymatic Synthesis of 5-Methyluridine
Using a Novel Combination of Nucleotide Phosphorylases. Biocatalysis and
Biotransformation 28: 245-253.
Gordon GER, Visser DF, Bode ML, Raseroka N.and Brady D (2011). A high
yielding multienzyme biocatalytic reaction for the synthesis of 5methyluridine by transglycosylation of guanosine and thymine. Journal of
Biotechnology, 151: 108-113.
Gordon GER, Visser DF, Bode M L, Lepuru J., Zeevaart JG, Ragubeer N,
Ratsaka M, Walwyn D and D Brady (2011). A chemo-enzymatic route for
preparation of β-thymidine, a key intermediate in the preparation of
antiretrovirals. Organic Process Research and Development 15: 258-265.
Visser DF, Hennessy F, Rashamuse J, Pletschke B and Brady D (2011).
Stabilization of Escherichia coli uridine phosphorylase by evolution and
immobilization. J. Mol. Cat. B: Enz, 68: 279-285.
MeO 2C
NHCbz
O
MeO 2 C
NHCbz
N
H
a
O
R R-9
O
N
H
O
RR-Monatin is 2000
fold sweeter than
sucrose.
HO 2C
RR/ SS-9
NHCbz
O
N
H
SS-12
O
Enzymatic resolution of RR/SS monatin derivatives. Reagents and Conditions: a) ChiroCLEC-BL, ACN, phosphate
buffer (0.3 M, pH 7.5) 22°C, 15-20 h.
MeO 2 C
HO2 C
NHCbz
a
O
RR-9
NH2
b
OH CO 2H
O
N
H
HO 2C
NHCbz
N
H
OH CO2 H
N
H
14
2
Reagents and Conditions: a) RR-9 in DCM, aq. KOH, 25 C, 30 min, then AcOH; b) 10% Pd/C (5%), MeOH, Et3N, H2 (3
bar), 40 C, 1 h.
© CSIR 2012
www.csir.co.za
Rousseau AL, Buddoo SR, Gordon GER, Beemadu S, Kupi BG, Lepuru MJ, Maumela MC, Parsoo A, Sibiya DM, and Brady D
(2011). A chemo-enzymatic process for stereoselective synthesis of monatin. Organic Process Research and Development
15: 249-257.
R1
R3
O
R2
* * R
4
Epoxide
hydrolase
R1
H2O
R3
O
OH
R2
+
R4
Starting epoxide
(2,3 -Epoxypropyl)benzene
R3
Blockbuster
R2
R1
OH
R4
2-benzylmorpholine
(appetite suppressant)
H
N
O
O
Racemic
epoxide
Optically active
chiral epoxide
Optically active
chiral diol
Styrene oxide
(S) -Fluoxetine (Prozac)
(anti-depressant)
C F3
O
O
N
H
p - Nitrostyrene oxide
(R) Nifénalol (beta blocker)
OH
O
NO 2
Chiral Epoxide use in Pharmaceuticals
N H iP r
NO 2
trans -1-Phenylpropene oxide
Ephedrine (stimulant)
OH
O
NH C H3
•
•
•
•
•
•
Cardiovascular agents
– beta-blockers
Antitussive drugs
Antibiotics
Antiviral agents
MAO inhibitors
Biochemical probes
© CSIR 2012
www.csir.co.za
Indene oxide**
O
Indinavir (Crixivan)
HIV protease inhibitor
C 6H 5
N
N
OH
C H2
N
C O N H C ( C H 3) 3
O
H
N
OH
DEPA
N
HO
Nitrile
hydratase
O
KCN
2-Hydroxy-3phenyl-propionitrile
Phenylacetaldehyde
OH
NH2
HO
O
HO
O
Amidase
2-Hydroxy-3phenyl-propionamide
2-Hydroxy-3-phenylpropionic acid
Cascade reaction:
α-hydroxynitriles from
aldehydes
Strand RF
Strand
Strand RF
RF
plus 1
plus 1
1
plus
plus 2
plus
2
plus 2
plus 3
plus
plus 3
3
1
1
1
198
198
198
397
397
397
595
595
595
cDNA LNHase beta alpha chap
Universal
code
cDNA
LNHase
beta
alpha chap
cDNA LNHase beta alpha chap
Universal code
code
794 Universal
993
1191
1390
794
993
1191
1390
794
993
1191
1390
1588
1588
1588
1787
1787
1787
1986
1986
1986
Chaperone
Beta
Alpha
minus 1
minus
1
minus 1
minus 2
minus
minus 2
2
minus 3
minus
3
minus 3
© CSIR 2012
www.csir.co.za
Brady D, Beeton A, Kgaje C, Zeevaart J, van
Rantwijk F, Sheldon RA (2004) Characterisation
of Nitrilase and Nitrile Hydratase Biocatalytic
Systems. Applied Microbiology and
Biotechnology. 64: 76-85.
•
•
•
•
© CSIR 2011
www.csir.co.za
Cross linked porous
polyethyleneimine (PEI)
matrix
Binding of functional
molecules throughout the
volume of microspheres.
PEI has high functional group
density.
These features translate to
exceptional binding capacity,
and enable microsphere
properties to be readily
engineered for performance
superiority.
•
Globally Sub-Saharan Africa is estimated as being one of the two regions with the
highest biomass (Smeets, 2004).
• SA has large sugar industry and
Pulp and paper industry.
• TIA project (jointly with
Universities) and COST action
Biorefining and Carbon cycling program,
University of Georgia (http://www.biorefinery.uga.edu/)
•
•
•
•
•
•
•
Industrial Biotechnology (general)
Bio-based Economy (BE-BASIC in NL)
Fermentation (Strong in NL)
Biocatalysis (Strong in NL)
Functional Protein Production
Enzymes
Bioremediation