for “natural label” flavors - bio kumbhar.pdf · advances in bio-catalytic transformations for...

Advances in bio-catalytic transformations

for “Natural Label” flavors

Pramod Kumbhar, Ph.D.

Chief Technology Officer

24th July 2017

© Praj Industries Ltd www.praj.net 2

Praj – India’s Leading Industrial Biotech Company

-Alcohol/ Ethanol plants

-Brewery plants

Water and Wastewater Treatment

Systems

Critical Process Equipment &

Systems

HiPurity Systems

- Pharma

- Biotech

- F&B

- Cosmetics

Bio-Products

-Distillery bioconsumables

-Livestock, health & nutrition

products

-Human health & nutrition

-Biochemicals

Quick Facts

GLORIOUS YEARS

SINCE INCEPTION

GLOBAL MARKET

SHARE IN ETHANOL

PEOPLE

CRORE

MANUFACTURING

AND FABRICATION

FACILITIES

FY16 REVENUES FROM

EMERGING BUSINESSES

YEARS

CONSISTENT

DIVIDEND PAYING

COMPANY FOR

HUMAN CAPITAL OF

MORE THAN

HOLDS

CONSOLIDATED

REVENUE IN FY16

GLOBAL REFERENCES

OVER COMPLETED

BUSINESS FROM

REPEAT CUSTOMERS

>50% MARKET SHARE IN

EMERGING MARKETS

FOR ETHANOL PLANTS

PATENTS

Praj Matrix – R&D center

US$ 25+ Million investment

80,000 sq ft of Labs, Pilot Plants, and Offices

90+ technologists

20 PhDs, 65 Masters

5 Technology COEs

Biology, Chemistry, Engineering

16 Well Equipped Labs

ISO-9001-2008 Analytical Labs

Pilot Plants

1 tpd Cellulosic Ethanol pilot plant

Multipurpose Pilot Plant

Recognized research center by DSIR, Govt of

India

4

Bench and Pilot scale facilities enable validation of scientific

assumptions and rapid commercialization

Biologists

Chemists

Engineers

Ph.D.: 08

Post-graduates:23

Ph.D.: 02

Post-graduates:15

Ph.D.: 01

Graduates & Post-

graduates:09

Ph.D.: 02

Post-graduates:18

Ph.D.: 06

Post-graduates:06

Centre of Excellence

Micro and molecular Biology Expertise

Classical Mutagenesis Directed Evolution Genome Shuffling Medium throughput microbial Screening Screening in deep well plates, shake flasks, 1 L fermenters Yeast cross-breeding Bioassay Development

Cloning & Expression in Bacteria and Yeast Gene Deletions Cofactor engineering Heterologous expression Protein secretion Targeted improvements for substrate utilization, by product reduction / elimination

Enzyme characterization Enzyme purification and formulation Enzyme performance evaluation Application Development Enzyme activity assay development Enzyme kinetic studies

Microbiology

Molecular Biology

Enzymology

7

© Praj Industries Ltd www.praj.net

Biochemicals Development at Praj

Product

Price

Product Volume (TPY)

100

10

2

$/k

g-

50

2-3 BDO

Levulinic

Acid

EPA+DHA

Natural Vit E

Phytosterols

Xylitol

1000

300

Hyaluronic Acid

Bacteriocins

Technology Capability Product Opportunities

Molecular & Micro-Biology

Process Development

Downstream Process

Chemical Sciences Furfural

Fragrance molecule 1

Fragrance molecule 2

Ethanol

Flavors & Fragrance Industry

• USD 25 Billion world market

• About 50:50 split between Flavors and Fragrances


* http://www.leffingwell.com/top_10.htm

USA (FDA) /EU flavor regulations


• Essentially same flavourings are allowed but designated differently

• US and EU define “natural” differently

US : An essential oil, oleoresin, essence or extractive, protein hydrolysate

distillate, or any product of roasting, heating or enzymolysis (source

material must be natural)

EU = Source material must be vegetable, animal or microbiological. Must be

produced by a traditional food preparation process (source material vague)

EFFA guidance document for “Natural” Flavorings

Regulations governing “natural” products specifically

permit “fermentation” and “Enzymatic/microbial”

processes

Praj Proprietary & Confidential

List of

permissible

processes

Accepted Principles for Preparation of “Natural”

Flavors


• Source materials must occur in nature

• Water and atmospheric air can be used during production

• When considering a process, the resultant material must be found in

nature or as an artefact in traditional foods

• In biotechnological processes, the substrate must be natural, but the

nutrients can be used regardless of source

• Processing conditions are allowed so long as they meet the

processing criteria of traditional or in--home preparation techniques

Why Natural?


• Survey indicates that almost 60% of

consumers in the United States look for

the word “natural” when they shop for

food products

• More than 65% of flavouring ingredients

used in US are labelled natural and

have food market potential of $ 9 billion

Pathways for preparation of “Natural” Flavor


Driving Forces for bio catalysis in Flavor production


Progress of Biocatalysis


Biocatalysis : Process Development


All “Natural” Raspberry ketone synthesis


Betuloside extracted from

birch bark

Hydrolyzed using b-Glucosidase

which is commercially

available

Fermentation with Candida

boidinii (secondary

dehydrogenase) to form

Raspberry ketone

Biotransformation : Synthesis of “Green Notes”


Its Vanilla World!

C& EN : Volume 94 Issue 36 | pp. 38-42

ISeptember 12, 2016

“The amount of all the vanilla beans in the world is not sufficient to flavor

everything that everyone wants to flavor with vanilla.”

—Carol McBride, U.S. vanilla category Symrise

Sensory experts perception

• Fairly good match

Retro-synthetic approach for Biocatalysis

• Target structure is subjected to

deconstruction

• Selection of microbes or enzymes

based on literature and techno-

economics

• Synthesis of intermediates and

final product


de Souza et al. Chemistry. 2017 May 17.

doi:10.1002/chem.201702235

Availability of Enzymes and consideration for

their use


• Broad variety of enzymes (including immobilised form) are now Commercial available

facilitating their direct as well as their simple recycling.

- hydrolases (lipase, esterase, protease),. ketoreductases, transaminases,

imine reductases, dehalogenases

• Several companies offer collections of enzymes in the microtiter plate format for rapid

screening.

• Cofactor-independent enzymes are more easy to use simply because the cofactor

need not be recycled

• Many enzymes tolerate the presence of organic solvents. As a rule of thumb,

concentration of water-miscible organic solvents (DMSO, DMF, lower alcohols etc.)

should not exceed 10-20 % (v/v).

Conclusion

• For the industrial flavor industry, the biotransformation platform offers

following advantages

Potential to secure more uniform, uninterrupted supplies of

high-value materials based on fermentation

Biosynthetic products manufactured via microbial fermentation

are deemed “natural” or “substantially equivalent” to a

botanically derived product

Lower cost and higher Purity

Sustainability

© Praj Industries Ltd www.praj.net 23 Praj-Matrix ready to provide end to end solution

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