Factors affect the enzymatic reaction and approaches

Enzyme concentration
Enzyme concentration used might not be sufficient
Amount of product formed over a specified period of time will be dependent
upon the level of enzyme present
The approach that can be taken is by designing the enzyme assay so that
the observed activity is proportional to the amount of enzyme present in
order that the enzyme concentration is the only limiting factor which is
satisfied only when the reaction is zero order. Enzyme activity is generally
greatest when substrate concentration is unlimiting.

Factors affect the enzymatic reaction and approaches

Presence of Inhibitors
May alter the catalytic action of the enzyme and consequently slow down the
catalysis.
Therefore, by increasing the concentration of the substrate, the substrate can
out-compete the inhibitor, and so the normal reaction can take place at a
reasonable rate.

Factors affect the enzymatic reaction and approaches

Temperature
Enzyme might be adversely affected by the unsuitable temperatures and
The selection of optimum temperature is required in order to ensure the activity is
increasing.

Factors affect the enzymatic reaction and approaches

pH
Extremely high or low pH values generally result in complete loss of activity for
most enzymes
It is the factor in the stability of enzymes.

The most favorable pH value is at the point where the enzyme is most active
which is known as the optimum pH

BIOCATALYTIC REACTION OPTIMIZATION

Strategies to achieve efficient biocatalytic reaction
Medium engineering:
Variations in solvent systems can influence the performance of enzymatic
transformations
Modification of the immediate vicinity of the biocatalyst can be done in non-aqueous
media.
The enzyme inhibition that might occur at high substrate levels in the process can also
be overcome by the addition of co-solvents.
Several aspects have to be considered in choosing an appropriate solvent for a given
reaction which include compatibility with the selected reaction (substrates and
products), inertness, low density to minimize mass transfer limitations, and other
properties that are suitable (e.g. surface tension, toxicity, flammability, waste disposal
and cost).

BIOCATALYTIC REACTION OPTIMIZATION

Strategies to achieve efficient biocatalytic reaction
Substrate engineering:
Use of modified substrates has been investigated to improve efficiency of
enzymatic reactions.
Easy separation of the substrate and the product can be achieved through
this approach.
This process has the advantages of easy scalability, a slightly higher
enantioselectivity compared to the reaction performed with vinyl acetate as
an acyl donor and a simple flash chromatographic separation.
Creating variations in the substrate structure has also been tried in order to
achieve improvements in biocatalysis.

BIOCATALYTIC REACTION OPTIMIZATION

Strategies to achieve efficient biocatalytic reaction

Biocatalyst engineering:
Enzymes are being modified.
Through variety of forms and formulations of enzymes have been
employed in non-aqueous systems: native enzymes, suspended
enzyme powder, solid enzyme adsorbed on support, polyethylene
glycol-modified enzymes soluble in organic solvents, enzyme
entrapped within a gel or microemulsion or reversed micelle and
immobilized enzyme.
The activation of enzymes can be done by adding an inorganic salt or
noncovalent modification of the native enzyme.

BIOCATALYTIC REACTION OPTIMIZATION

Strategies to achieve efficient biocatalytic reaction

Biocatalyst discovery and optimization:

Widely used to identify genes from different organisms and
genome-wide comparisons through the modern tools of
screening, high throughput screens, and availability of protein
sequence databases and bioinformatics in screening for novel
biocatalysts.
Mathematical and data-mining tools also have now been applied
to analyze sequence-activity relationships of peptides and
proteins and to assist in the design of proteins and peptides with
specified properties.