Innovationbyevolution:Bringingnewchemistrytolife

Frances H. Arnold California Institute of Technology

May 18, 2017

Imag

es c

ourte

sy o

f Shu

tters

tock

Fe

O

SCys

+�

Fe

C

SCys

RR

Fe

N

SCys

R

Carbenetransfer

Nitrenetransfer

X

AATAGCCGTTATTTCCGGATGTGCATAGCTGATTTGACCCATCCGGTACACCAATDACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGTTAACCACACGTCCCGGATATATTTAATTGGCCGGAGAGTCTCCCGCGCGACATAAGGAGTCCTCGTTTCGAGATACGTACGGCATGGTGACACCAGTTGCCCTCTGATTCCCGGAGCCTCTTTGAAAACGTCGAGTCGAATCGAAGTTCGAACCCCGGATCGGGTCCACCAACTTAGAGATGTGTGTGCGCTGACTCAGTCATTTTGACCCATCCGGTACACCAATGGGTCGGACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGTTAACCACACGTCCCGGATATATTTAATTGGCCGGAGAGTCTCCAATAGCCGTTATTTCCGGATGTGCATAGCTGATTTGACCCATCCGGTACACCAATGGGTCGGACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGT

AATAGCCGTTATTTCCGGATGTGCATAGCTGATTTGACCCATCCGGTACACCAATDACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGTTAACCACACGTCCCGGATATATTTAATTGGCCGGAGAGTCTCCCGCGCGACATAAGGAGTCCTCGTTTCGAGATACGTACGGCATGGTGACACCAGTTGCCCTCTGATTCCCGGAGCCTCTTTGAAAACGTCGAGTCGAATCGAAGTTCGAACCCCGGATCGGGTCCACCAACTTAGAGATGTGTGTGCGCTGACTCAGTCATTTTGACCCATCCGGTACACCAATGGGTCGGACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGTTAACCACACGTCCCGGATATATTTAATTGGCCGGAGAGTCTCCAATAGCCGTTATTTCCGGATGTGCATAGCTGATTTGACCCATCCGGTACACCAATGGGTCGGACAAATCCCGATTTGATCGTGTGCGCGACATGTCTTCCGGCGACACATGTGTCTCTCACTCCGAGAGATCGGTTAGAGTCTCGGT

MTIKEMPQPKTFGELKNL...KETSPIPQPKTFGPLGNL...KQASAIPQPKTYGPLKNL...WRRRGIPGPLGYPLVGSF...WIRKGVKGPRGLPFLGVI...FIRKGIKGPRGFPGIGML...WIRKGVKGPRGFPFFGVI...WIRKGVKGPRGFPFFGVI...WMRKGIKGPRGLPFFGII...WMRKGVKGPRGRPFVGVL...WRRRGVVGPMGFPVLGVF...REKIGLSGPEPHWFLGNL...REKIGLTGPEPHWFMGNL...RSSIGIPGPPVHWLWGNL...KVSKYPKGPLPLPFIGNI......

Enzyme families are the products of millions of years of mutation and natural selection.

mutationandselection

Ancestralenzyme

Evolution does it very well.

Each molecule is well adapted to its biological role.

Nature does it in real time: atrazine

• 1950–1993 non-biodegradable; accumulated in soil• From 1993 onwards, fast degradation observed

N

N

N

Cl

NH

NH

the internetofliving things

Crowd-sourcing problem solving for 3+ billion years

Humans have altered the biological world using evolution by artificial selection

for thousands of years.

Natural mechanisms for generating sequence diversity are limited

Not so in the test tube…

Imag

e co

urte

sy o

f Shu

tters

tock

Sex (DNA recombination) is effective strategy for creating new sequences and functions.

MTIKEMPQPKTFGELKNL...KETSPIPQPKTFGPLGNL...KQASAIPQPKTYGPLKNL...WRRRGIPGPLGYPLVGSF...WIRKGVKGPRGLPFLGVI...FIRKGIKGPRGFPGIGML...WIRKGVKGPRGFPFFGVI...WIRKGVKGPRGFPFFGVI...WMRKGIKGPRGLPFFGII...WMRKGVKGPRGRPFVGVL...WRRRGVVGPMGFPVLGVF...REKIGLSGPEPHWFLGNL...REKIGLTGPEPHWFMGNL...RSSIGIPGPPVHWLWGNL...KVSKYPKGPLPLPFIGNI......

Sex with molecules

Performance

0

10

20

30

40

50

60

70

0 1 2 3 4 5 6 7

Gen1 2 3 4 5

Directed evolution (iterative mutation and artificial selection) can re-optimize biological molecules for new functions.

Evolved protein doing a new job

Native protein doing its native job

Native protein doing a new job

Second most abundant element in the Earth’s crust

Si14

28.0855Silicon

...the literature is void of examples of biologically synthesized, either in vivo or in vitro,

silicon-carbon bonds.Silicon 2009, 1, 147.

NH

SiOHR

R

catalysts / ligands

SiR3reagents for cross-coupling

Si

18F-acceptors for PET imaging

Si(i-Pr)3

Si(i-Pr)3materials for organic LED

biological probes and drug leads

O

NH

H2N

Me3Si O

OH

18FRR

3

Human chemists make lots of things with silicon-carbon bonds

lubricants, sealants, coatings…

Rhodothermus marinus cytochrome c

Si H

MeMeN2

Me

O

OEtMe

O

OEtSiMeMeRma cyt c

+

• Made by a gram-negative, thermohalophilic bacterium• small protein – 124 amino acids• Tm = 106 °C

Directed evolution converted this protein into a highly active catalyst for silicon-carbon bond formation…

0

500

1000

1500

2000

WT M100D V75TM100D

V75TM100DM103E

TTN

97%ee

>99%ee

>99%ee

>99%ee

(act

ivity

)

...inside living cells.

70+ employees at Gevo Headquarters in Englewood, CO

30+ employees at Gevo Plant in Luverne, MN

Publicly Traded Since Feb. 2011(NASDAQ Symbol GEVO)

We founded Gevo, Inc. in 2005

Gevo,Inc.,theworld'sonlycommercialproducerofrenewableisobutanol,announcedthattheU.S.ArmyhassuccessfullyflowntheSikorskyUH-60BlackHawkhelicopterona50/50blendofGevo'sATJ-8(Alcohol-to-Jet).

December, 2013:Gevo Supplies U.S. Army With ATJ-8 Fuel for the Black Hawk Helicopter

Gevo,Inc.,theworld'sonlycommercialproducerofrenewableisobutanol,announcedthattheU.S.ArmyhassuccessfullyflowntheSikorskyUH-60BlackHawkhelicopterona50/50blendofGevo'sATJ-8(Alcohol-to-Jet).

November 15, 2016:

Pest Control asNature Intended

28 ©2017Provivi,Inc. CONFIDENTIAL28

MATINGDISRUPTIONTHENEWSTANDARDFORINSECT CONTROL

01

Founded in 2014:

Science & Environment

Fall armyworm 'threatens African farmers' livelihoods'By Helen BriggsBBC News

6 February 2017 Science & Environment

The armyworm burrows into cobs

Scientists are calling for urgent action to halt the spread of a pest that is destroying maizecrops and spreading rapidly across Africa.

The fall armyworm poses a major threat to food security and agricultural trade, warns the Centre for

CABI

News Sport Weather Shop Earth Travel

FallarmyworminvadedAfricain2016

• Fall armyworm & corn earworm mating disruption• Main pest of fresh sweet corn in Florida (30 sprays!)• $600M spent in Brazil corn just for fall army worm

First product: sweet corn in 2018

Preventative control with mating disruptionArgentina field trials in 2016

Conventional practice

10 insecticide sprays Only 3 insecticide sprays!

0

10

20

30

40

50

60

70

80

24.heinä 3.elo 13.elo 23.elo 2.syys 12.syys 22.syys 2.loka 12.loka

%FA

WIn

festation

Provivi

Provivi (98% mating disruption)

Conventional grower practice

Jane Goodall

Diversity of components experiencesminds