Dinesh Nair; Novozymes

BIOCATALYSIS- AN EFFECTIVE TOOL FOR A

COST EFFICIENT,SUSTAINABLE &

GREEN MANUFACTURING

December 07, 2013

The World Leader in Bioinnovation

• Business consists of two

divisions: Enzyme Business and

Bio Business

• 2011 Sales of USD ~ 2.1 billion

• 2011 EBIT margin of ~ 22.3%

• About 14% of our revenue is

spent on research and

development

• More than 700 products used in

130 countries in over 30 different

industries

• 6,000+ granted and pending

patents

• 5,200+ employees

Founded 1941

Listed 2000

Ticker NZYM

Exchange Copenhagen

BioBusiness

Technical enzymes

Foodenzymes

Feedenzymes

Micro-organisms

Biopharma

Enzyme Business

Detergent enzymes

Industrial cleaning

Wastewater treatment

Bioagriculture

Pharmaceuticals & fine chemicals are one of the most polluting industries

IndustriesProduction Scale

(Tons per Year)E-factor

Oil Refining 106 - 108 < 0.1

Bulk Chemicals 104 - 106 < 1 - 5

Fine Chemicals Industry 102 - 104 5 - > 50

Pharmaceutical Industry 10 - 103 25 - >100

Source: R.A.Sheldon, Chem & Ind, 1992, 903 ; 1997, 12, IGCW 2011

Environmental impact of some of the chemical industries

3

A sustainable and greener footprint in chemical production is increasingly becoming a global need…

There are existing and emerging solutions to counter pollution

Improving Manufacturing Sustainability: An Overview

IN (kg)

ProductPROCESS

Raw Material 1

Raw Material 5

Raw Material 2

Raw Material 4

Raw Material 3

Out (kg)

LOSSES WasteReject

By-products

Re-cycle

Re-use

Down-cycle

Re-sale

£

££

Pharmaceutical manufacture typically has > 100 kg raw

material consumption/kg product• Disposal costs

• Infrastructure costs

• Energy costs

Source: GSK presentation

Biotransformation is the sustainable answer to growing concerns of chemical industry

Biocatalysis (Isolated Enzymes)Whole Cells

Fermentation

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Biodegradable

Renewable sources

Nature’s Catalysts

There are several benefits of using enzymes over chemical reaction to produce APIs & Fine Chemicals

General Advantages using Enzymes in API/Fine Chemical Manufacturing

Improved Productivity− Higher Yields − Shortened synthesis route

Milder reaction conditions− Mostly ambient reaction condition

Greater Savings Potential− Replaces costly chiral resolving agents − Significant reduction in waste streams

Greater Selectivity− High level of Stereo-, regio-, and

chemo-selectivity

Simplified work-streams− Simplified processing and purification − Fewer byproducts, reducing impurities

6

Biocatalysis reactions adhere to green chemistry principles

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1. Enables new, sustainable

routes reducing waste

2. Enables more efficient routes

3. Generally low toxicity

4. Enables designing safer

molecules

5. Water as solvent or Class

II/Class III solvents

9. Catalytic with sustainable

and greener foot print

10. Flexibility in process design

and degradable

11. Real time analysis on

impact on pollution

possible

12. Minimal (negligible) risks

associated with reactions

8. Chemo, regio & stereo-

selective in nature

6. Performed at ambient

conditions7. Renewable

Parameters of green chemistry principles in relation with biocatalysis

Biocatalysis gets its principles from several disciplines and applies to a number of industries

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Biocatalysis

Analytical Medicine

Pharmaceuticals

Food & Nutraceuticals

Animal Feed

Crop Protection

Pulp & Paper

Mining

Specialty Chemicals

Microbiology

Molecular Biology

Biochemistry

Organic Chemistry

Process Chemistry

Reaction Engineering

Reactor Designing

Kinetics

Contributing Disciplines Application Areas

Based on the reaction types, enzymes are grouped into several classes

Selected Reactions Enzyme ClassEC No.

Key Applications

C=O and C=C reduction

Reductive amination of C=O

Oxidation of C-H,C=C, C-N and C-O

Cofactor reduction/oxidation

Oxidoreductase 1

Atorvastatin

Intermediate

Montelukast

Transfer of functional groups such as

amino,Acyl,phosphoryl, methyl, glycosyl, nitro

& sulphur groups

Transferase 2

Cyclodextrin from

Starch

Sitagliptin

Hydrolysis of esters, amides, lactones, lactams,

epoxides, nitriles & reverse reactionsHydrolase 3

Diltiazem

Intermediate

Resolution of rac.

phenylethylamine

Addition of small molecules to double bonds

such as C=C, C=N and C=O

Lyase

(synthase)4

L-Malic acid

Statin Intermediate

Isomerisations such as racemizations,

epimerizations & rearrangement reactionsIsomerase 5 HFCS from glucose

9Source:

Some key examples of enzyme classes and their applications

Hydrolases are the most used enzyme class in biotransformation's followed by oxidoreductases

Biocatalysis is a proven technology for some prominent active pharmaceutical ingredients synthesis

Pregabalin- Lipase from TL Source

Sitagliptin- Transaminases

Simvastatin- Acyltransferases

10

Few important case studies on successful chemo-enzymatic synthesis

Pregabalin Biocatalytic Route

Pregabalin Chemical synthesis Pregabalin Biocatalytic Route

11

Improvements due to replacing chemical process with chemoenzymatic reaction

InefficientResolution at rac.pregabalin

stage

Novozymes Lipase

Lipozyme TL 100 L

Flow Chart of Pregabalin Process……..

12

(R)-Diester

(S)-Diester

Water

Enzyme

(R)-Diester

(S)-Monoester

Water

Enzyme

(S)-Monoester

Water

Enzyme

Water

Enzyme

Racemisation

Recycling

Key Advantages of Biocatalytic Route

Low protein Loading(0.8%)

Resolution at first stage-wrong isomer can be recycled

High throughput

All reactions conducted in water

E-factor improved from 86 to 17

Starting material reduction by 800 Mt

(cumulative)

Drastic reduction in solvent usage

Mandelic acid usage-500Mt eliminated

(Cumulative)

Energy usage reduced by 83%

Pregabalin

Biotreatment

23%

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Inputs Chemical Route

Kg

ChemoenzymaticRoute

Kg

CNDE 6212 4798

Enzyme 0 574

S-Mandelic acid 1135 0

Raney Nickel 531 79.5

Solvents 50042 6230

Total 57920 11681.5

85%

88%

Reduction

Pregabalin Chemoenzymatic route used >5x less inputs than Chemical Route

Raw Material Inputs for 1000kg API Output

Source: Organic Process Research & Development 2008, 12, 392.

Key Advantages of Biocatalytic Route

10-13% increase in overall yield.

53% increase in productivity (kg/L,

per day).

19% reduction in total waste.

The elimination of heavy metal use

Reduction in total manufacturing

cost.

Sitagliptin Biocatalytic Route

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Improvements due to replacing chemical process with chemoenzymatic reaction

Source: Science, vol. 329, ,July 2010..

200g/L Substrate Loading

6 gm/L Enzyme Loading

50% DMSO-water system

Key Features of Biocatalytic Process

Simvastatin biocatalytic route drastically reduces number of steps from 6 to 2.

Simvastatin Biocatalytic Route

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Simvastatin Chemical synthesis Simvastatin Biocatalytic Route

Improvements due to replacing chemical process with chemoenzymatic reaction

Source :Current Opinion in Chemical Biology 2009, 13:43–50

Future outlook of biocatalysis is very promising

Directed Evolution

Variant Selection or screening

Smarter Libraries of Enzymes

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Advanced and Emerging Technologies

DNA technology & Protein engineering

Bioinformatics

Molecular Modelling studies-Docking of

substrate with proteins

Reaction engineering & process intensification

using biocatalysts

High throughput enzyme screening

Enzyme screening kits

First wave

living cells applied to useful chemical

transformations

Second Wave

Initial protein engineering techniques

extended range of enzymes to allow

synthesis of unusual synthetic

intermediates

Biocatalysis evolution so far and future outlook

So Far… the first and second wave

The third wave: future outlook

The way forward……….

Biocatalysis likely to be one of the most important tools for Fine Chemicals & Pharma for

cost effective & sustainable manufacturing

− It is likely that biocatalysts would be available for all types of transformations in future

− Tailor made enzymes by directed evolution & random mutagenesis

More and more collaborations between fine chemicals/pharmaceutical companies & Enzyme

companies are expected to take place to solving complex industrial problems

Biocatalysis companies will likely evolve into companies providing end to end solutions from

screening of variants to optimizing hits to process optimization to intermediate

manufacturing

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… And Novozymes is adequately positioned and aggressively poised to help the industry in this journey

let’s talk to find out how we can work together

CONTACT

NAME : DINESH NAIR

TITLE : REGIONAL BUSINESS

MANAGER

EMAIL : DINN@NOVOZYMES.COM

WWW.BIOPHARMA.NOVOZYMES.COM

Thank you for attention