CarbaZymes KET 2016

15 Pages·2017·9.78 MB·English

by  Agapi Theodoridou

Maximizing the Impact of KET Biotechnology, EC-Workshop Brussels 2016
Sustainable
 industrial
 processes

based
 on
 a
 C–C
 bond-­‐forming

enzyme
 plaNorm
Wolf-­‐Dieter
 Fessner/TUDA
 –
 Simon
 Charnock/PROZO

This
 project
 has
 received
 funding
 from
 the
 European
 Union’s
 Horizon
 2020

Research
 and
 Innova>on
 Programme
 under
 Grant
 Agreement
 No
 635595
C-­‐C
 Bond
 Forma>on
 in
 Pharma
 Patents

Evolution of C–C bond formation over time
cross coupling chemistry
mostly Pd catalysis
boronic acids, stannanes
J. Med. Chem. 2016, 59, 4385
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Palladium
 Mining

Norilsk
Norilsk has largest known Ni-Cu-Pd deposits in the world.
Norilsk is one of the 10 most polluted places on Earth.
1% of global SO emissions are from Norilsk's Ni mines.
2
Estimated 4,000,000 tons of Cd, Cu, Pd, Ni, As, Se,
and Zn are released into the air every year.
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C-­‐C
 Bond
 Forma>on
 by
 Palladium
 Catalysis

The toolkit for C–C bond formations looks much different from 40 years ago:

In 2015 the preferred types are Suzuki, Sonogashira, Stille [...] reaction [...].
The Pd-catalyzed reactions are highly selective, mechanistically well-understood
and operationally easy to run.

On the contrary, in 1976 the most used reaction types were Wittig olefination,
Grignard, Barbier, Friedel–Crafts acylation, Mannich, Horner–Wadsworth–Emmons,
aldol and Knoevenagel condensation. All of these depend upon modifying the reactivity
of a carbonyl group to form the new C–C bond: most use strong bases (pK > 20)
a
or strong Lewis acids (AlCl , BF ) as promoters.
3 3
reaction types preferred by Nature
mild conditions due to enzyme catalysis
no strong base needed
no strong acid needed
4
C-­‐C
 Bond
 Forma>on
 by
 Enzyme
 Catalysis

Nature‘s toolkit for C–C bond formations offers multiple opportunities
O OH
O S. cerevisiae
+
CO H
2 CO O
2
(R)-PAC ⇒ Ephedrin
HO C
2
CO H
HO 2
CO H
2 O O
aldolase
+
O monatin
N CO H
2 N
H precursor
H
O O DERA OH OH O O OH
Cl
+
Cl Cl
statin
OH precursor
5
CarbaZymes
 Project

construction and use of innovative, competitive platform technologies
development of cutting-edge biotechnology as future innovation driver
widening industrial application of enzymatic processes
6
CarbaZymes
 Impact:
 Enzymes

ALD (024)
enzyme panel construction:
from hits to homologs
7
CarbaZymes
 Impact:
 Bioinforma>cs

3DM
Protein
 Super-­‐family

Informa>on
 Systems

(cid:18535)  Structures
(cid:18535)  Alignments
(cid:18535)  Literature
(cid:18535)  All the data is
connected
8
Mul>-­‐scale
 Enzyme
 Produc>on

How
 to
 produce
 1000
 novel
 enzymes!!!

9
CarbaZymes
 Impact:
 Innova>on

Science Technology Process Market

Research Development Demo Market

IP know-how Product Return
Milestones •  large-scale bioprospecting (>500 novel aldolases)
achieved •  3DM superfamily databases constructed for enzymes of interest
•  focused libraries constructed for enzyme optimization
•  thermostable variants of enzymes of interest found and characterized
•  mutability landscapes constructed for activity, chemo-, solvent,
thermostability
•  supercompetent Thermus strain constructed
•  biocatalytic process development for scale-up
•  transfer in progress for new developments
into operational environments
mid-term •  1 patent application filed (pending)
status •  2 patent applications in preparation
•  several spin-offs: new chiral building blocks for future applications
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