Biocatalysis Position

In Green Chemistry

http://www.nature.com/scitable/topicpage/cell-metabolism-14026182

How Green is Biocatalysis? To Calculate is To know

Yan Ni, Dirk Holtmann and Frank Hollmann
ChemcatChem 2014, 6, 930-945

DEFINITIONS AND CONCEPTS

Organic synthesis concept

Organic synthesis is concerned with the construction of
molecules, involving chemical reactions. Several reactions
will take place one after another (or in parallel) until the
targeted molecule is complete.
A chemical reaction model:

+
Molecule 1

+
Molecule 3

Molecule
Molecule
Molecule343

Molecule 2

Molecule 5
Molecule 6

Example
Boots process for ibuprofen (analgesic like
aspirin) synthesis
(1960) : 6 steps and numerous non
recyclable waste.

Green Chemistry
Reduce the impact of chemistry on the environment by preventing pollution at its source
and using fewer natural resources

1990

U.S. Environmental Protection Agency (EPA)

Green chemistry efficiently utilizes (preferably renewable) raw materials,

eliminates waste and avoids the use of toxic and/or hazardous reagents
and solvents in the manufacture and application of chemical products.

1998
12 principles of green
chemistry by P. Anasta
and J. Warner

2007
REACH European rules : to secure production and utilisation of chemicals

12 principles of Green Chemistry

Avoid auxiliary
substances
Catalysts

Innocuous
degradation
products

Non-toxic
substances

Atom economy

Efficient
production

Green
Chemistry

Avoid
derivatisations

Preventing
waste

Diminishing
accidents

Real time
measurement

Save energy
Renewable
feedstock

Example
Green process

New process by BHC

(1990): only 3 steps and
only one sub-product
(acetic acid) which is
recycled for another use.

Biocatalysis
"The vast number of chemical
reactions taking part in a living
cell are controlled by organic
catalysts. Life is the
orchestrated combination of
processes catalyzed by enzymes."

WILLSTTTER RICHARD - (1872-1942)

Nobel price in 1915 chlorophyll and
other plant pigments

Biocatalysis may be broadly defined as the use of enzymes or

whole cells as biocatalysts for industrial or academic synthetic
chemistry.

Nature, a molecule architect?

Enzyme ubiquity and diversity
Escherichia coli

Human

4500 genes
1518 enzymes
1700 reactions
~ 25000 genes
~ 20000 proteins
~ 15000 enzymes

Nature, a molecule architect?

Prokaryotic cell

Eukaryotic cell

Orchestration? Metabolic map

Huge diversity of natural

Compounds and reactions

How this can work?

CHEMOSELECTIVITY, REGIOSELECTIVITY, STEREOSELECTIVITY
An average protein would contain 300 aa and as 20 proteinogenic amino acids
exist, 20300 (10390) different polymers are theoretically possible The complete
Universe is insufficient to contain a copy of each protein. An enzyme is thus
able to bind a substrate specifically to lead to a chemical reaction.
To give an idea: our Universe would contain 1080 atoms in the eye accessible part

Enzymes are chemoselective, regioselective and stereoselective

Chemoselectivity: reaction on a polyfunctional molecule possible without any protection.

Regioselectivity: Identical functional groups distinguished depending on their position on the
molecule.
Stereoselectivity: enzymes are chiral catalysts. They are able to differentiate stereoisomers and in
particular enantiomers.

From gene to protein

From gene to protein

Enzymes are proteins

Proteins are polymers (macromolecules) built up from amino acids.

Amino acids

Dipeptide

Oligopeptide

Protein

Organisation and folding of this chain lead to a 3D structure.

Enzymes are proteins

cristal
Cartoon representation

Soybean protein

Enzymes at work
They cut molecular assemblies into smaller molecules

They stick for constructing molecular assemblies

Enzymes at work
Key and lock concept

http://biosciences.dupont.com/about-us/guide-to-industrial-enzymes/

Enzymes at work

http://science.halleyhosting.com/sci/ibbio/chem/notes/chpt8/chpt8.htm
http://culturesciences.chimie.ens.fr/dossiers-reactivite-catalyse-autresdocs-Enzymes_Laage_1.html#menu
http://www.classhelp.info/Biology/AUnit3Biochemistry.htm

Enzymes at work

Rate factor increase from

107 to 1017 depending on
the enzyme

http://www.nature.com/scitable/topicpage/protein-function-14123348

Enzymes production
How industrial enzymes are made? And in the lab?

http://www.geni.org/globalenergy/library/technical-articles/generation/futurefuels/iogen/enzyme-technology/index.shtml

How enzymes are used?

In water, buffer, low temperature, mild conditions

However Enzymes are denatured by a change in temperature, pH or chemical

action

inactive

http://www.rsc.org/learn-chemistry/resources/chemistry-in-your-cupboard/vanish/8

APPLICATIONS
Detergent, food, pharmaceutical, chemical and leather industries
are producing and/or using enzymes

In food? Vanillin
Industrial synthesis, artificial aroma

(From lignin)

https://chempics.wordpress.com/category/industrial-chemistry/

Enzymes at home?

http://www.teknoscienze.com/Articles/HPC-Today-Detergent-enzymes-ndash-from-discovery-to-product-Thepower-of.aspx#.U3eWhXJPHqM

Amylase

Cellulase

Protease

In cars? Bioethanol

CELL FACTORIES

Microorganisms Tailored to the Raw

Materials Available
Ethanol is produced from non-food
biomass such as farming and forest
waste, green waste and dedicated
crops. This involves extracting the
cell wall (cellulosic) sugar from
biomass utilizing enzymes that
convert cellulose into sugar (glucose)
through hydrolysis. Then yeast strains
convert the sugars into ethanol via
fermentation. The resultant ethanol
is distilled to ensure it is of suitable
quality for use as biofuel. For this
biotechnology
process
to be
profitable, more efficient enzymes
are needed, which is a core focus of
R&D in this area

http://total.com/en/energies-expertise/renewable-energies/biomass/projects-achievements/futurol

Categories of chemicals produced by cell factories

http://www.nature.com/nchembio/journal/v8/n6/fig_tab/nchembio.970_F1.html

How to get efficient desired enzymes?

De novo design
(computer modelling)
Biodiversity

Efficient biocatalysts for

fine green chemistry or
industrial chemistry

Directed evolution of
existing enzymes

Directed evolution
Desired biocatalyst

??

Wild-type biocatalyst

Directed evolution
NEW GENES => NEW ENZYMES WITH NEW PROPERTIES => SCREENING
Evolution

Molecular biology

Screening

From biodivsersity
Harvest the DNA (genes) from
the environment

Databases

Screening for the desired properties

New enzymes
http://www.uniprot.org
http://www.rcsb.org/pdb/home/home.do
http://biology.unm.edu/biology/maggieww/Public_Html/ANALYSIS.HTM (list of databases)

CONCLUSION

12 principles of Green Chemistry

Avoid auxiliary
substances
Catalysts

Innocuous
degradation
products

Non-toxic
substances

Atom economy

Efficient
production

Green
Chemistry

Avoid
derivatisations

Preventing
waste

Diminishing
accidents

Real time
measurement

Save energy
Renewable
feedstock

BIOCATALYSIS HAS A PRIVILEGED POSITION IN GREEN CHEMISTRY!