transforming industries with high performance enzymes pac rim 2013...enzyme discovery at verenium...

David P. Weiner, Ph.D.

December 10, 2013

Transforming Industries

With High Performance

Enzymes

Global Challenges & Opportunities

• Unprecedented challenges to planet’s resources and environment

• Energy, Food, Water, Chemicals & Materials, Climate

• Opportunity for biotechnology to contribute sustainable solutions

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Current World Population:

© 2013 Verenium Corporation

The Power of Enzymes

• High catalytic efficiencies

• Exquisite selectivity – enantio-, chemo-, regio-

– Direct access to products

• Environmentally friendly

– Natural, biodegradable

– Reduce or eliminate waste streams

• Evolvable


Enzymes – Products in Our Daily Lives

Detergents

& cleaners

Human

Nutrition

Personal

care

Pharmaceutical

Animal health

& nutrition

Ethanol & other biofuels

Industrial


Enzyme Discovery at Verenium

Verenium’s Direct Approach

Bypasses Need for Culturing

TRADITIONAL

APPROACH

Typical

culturing

methods

access <0.1%

of biodiversity

Screen

Clone

Culture

Microbes

VERENIUM’S APPROACH

Extract Microbial DNA

Construct eDNA Library

High Throughput Screens

Faster, Comprehensive, Captures Nature’s Diversity


Unprecedented Microbial Biodiversity

• Over 2000 eDNA libraries

• Encompass the genomes from over 1 million

organisms

• Billions of genes, billions of potential new enzymes


Provides Powerful Competitive Advantage

Verenium’s Unique Suite of Technologies

Enzyme Discovery

Ultra-high Throughput

Screening

DirectEvolution®

Technologies

Expression,

Scale-up &

Manufacturing

Applications


From the Deep Blue…

to the Anthropogenic Abyss


• Screened Verenium’s extensive

environmental DNA libraries

• Discovered multiple candidates

• Applications work identified ideal enzyme

for fracturing

• Scaled-up & commercialized as

Pyrolase® cellulase

Development of Pyrolase®

Cellulase

Capturing The Best From Nature


(~85oC)

(~110oC)

Increased temperature range and pH while

maintaining unique performance properties of parent

enzyme

Improving The Best From Nature

Development of Pyrolase® HT Cellulase


α-amylase glucoamylase

Opportunity for new enzyme

HIGH TEMP, LOW pH

Development of a Novel Alpha-Amylase for

Starch Liquefaction

Higher activity

Higher temp

optimum

Lower pH

optimum Unmet Need


Discovery of Amylase Lead Enzymes

PE

RF

OR

MA

NC

E

PERFORMANCE THRESHOLD

pH, Temp, Activity

ENVIRONMENTAL

SAMPLES

INITIAL SCREEN SECONDARY

SCREEN

Millions of

potential

enzymes

15 enzymes

3 lead

enzymes

Guaymas Basin

Deep Pacific

Mediterranean Sea


Verenium’s GeneReassemblySM Strategy

3 parental genes (enzymes)

DIVERSIFY

BLEND

HIGH THROUGHPUT SCREEN

9 segments synthesized for each gene (8

points of demarcation)

39 combinations =

19,683 daughter constructs

Low pH activity, high specific activity and

expression

Best gene selected


From a Different Branch on the

Phylogenetic Tree of Life

Different Mode of Action

Produces More Uniform Substrate

Fuelzyme® Alpha-Amylase is Differentiated

from the Competition by Nature Itself

Most

α-amylases

Fuelzyme®

alpha-amylase

Superior thermostability, broader pH range & unique mode of action


Verenium Esterases & Lipases

• Over 600 unique esterases & lipases discovered

• Wide variety of useful properties including:

– Diverse fatty acid selectivities

– Broad range of temperature and pHs

– Activity on wide range of substrates

Key:

Verenium lipases

Commercial lipases


Photo courtesy of Prof. Greg Rouse, SIO

Development of Enzymatic Process for

Healthier Edible Oils

• Selectively remove unwanted fatty acid

• Upgrade heath profile, upgrade value

• Alternative to plant breeding / transgenics

+


Discovery of a Highly Selective Lipase

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10%

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90%

100%

1 2 3 4 5 6 7 8 91

01

11

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31

41

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91

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31

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FA #5

FA #4

FA #3

FA #2

FA #1

• Unprecedented range of enzyme selectivities from Nature

• GC screen identified enzyme with selectivity to FA #4

• Provided starting point for further optimization


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UHTP Screen (100,000 clones)

Evolved Variants

Wild type

Constructed

GSSMSM

Library

(~400 residues)

21

Optimization of Lipase Selectivity Evolving the Best from Nature

UHTP Screen (All possible combinations)

No Enzyme

Wild Type

Evolved

• Exquisite selectivity achieved (>99%)

• Additional round of evolution enhanced

thermostability

• Exceeded all performance targets


Industrial Enzymes: Profitable, Growing Markets

Today, New Markets Emerging


Source: Market size estimates based on Verenium internal estimates and L. Hepner & Associates Industrial Enzymes Report published in March 2013.

New Enzyme

Markets


Verenium’s Enzyme Collection

4,000+ Unique Proprietary Enzymes For Future Products

Betamannosidases

Cellobiohydrolases

Cellulases

Fucosidases

Galactanases

Glucanases

Mannanases

Pectinases

Pullulanases

Xylanases

Nitrilases

Phosphatases

Proteases

Amidases (primary)

Amidases (secondary)

Dehalogenases

Epoxide hydrolases

Esterases

Lipases

Phospholipases

Alphafucosidases

Alphagalactosidases

Alphaglucoronidases

Amylases

Arabinofuranosidases

Betaglucosidases

Isomerases

Aldolases

Ammonia-lyases

Ammonia-lyases (Tryptophanases)

C-O-lyases

C-O-lyases (Pectate lyases)

Dehydratases

Miscellaneous lyases

Catalases

Dehydrogenases

Oxidases

Transaminases

Nature does not hurry, yet

everything is accomplished -- Lao Tsu, 600BC “

“


• Strong foundation of technologies linked to business strategy

• Enzymes will be a key driver of the growing biobased economy

• Discovery together with evolution is expanding the role of enzymes

Looking Ahead to 2050


Dedicated to the memory of Patti Kretz and Patrick McCroskey

Acknowledgements

The Verenium team past & present

Our academic collaborators

Our industrial partners & customers

Through our unique access to nature’s ever-evolving

biodiversity, we create eco-friendly enzyme products that

improve performance and lower costs for today’s

global industries

Verenium is “Industry, Evolved.”


transforming industries with high performance enzymes pac rim 2013...enzyme discovery at verenium...

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