Bio-transformations

Microorganisms possess the capability to enzymatically modify a wide range of organic

compounds. Bio-transformations (bi-conversions or microbial transformations) broadly
refer to the processes in which microorganisms convert organic compounds into
structurally related products. In other words, biotransformation deals with microbial
(enzymatic) conversion of a substrate into a product with limited number (one or a few)
enzymatic reactions. This is in contrast to fermentation which involves a large number
reaction.

Although there are hundreds of bio-transformations known, only a selected few of them
are useful for the synthesis of commercially important products. The significance of
bioconversion reactions becomes obvious when the production of a particular compound
is either difficult or costly by chemical methods. Further, bio-transformations are
generally preferred to chemical reactions because of substrate specificity, stereo specificity
and mixed reaction conditions (pH, temperature and pressure).

Types of Biotransformation Reactions

Many types of chemical reactions occur in bio-transformations. These include oxidation,

reduction, hydrolysis, condensation, isomerization, formation of new C—C bonds,
synthesis of chiral compounds and reversal of hydrolytic reactions. Among these,
oxidation, isomerization and hydrolysis reactions are more commonly observed in bio-
transformations. Many a times bio-transformations involve more than one type of
reaction.

The conversion time required for biotransformation is related to the type of reaction, the
substrate concentration and the microorganism used. In general, oxidation, hydrolysis
and dehydration reactions are completed in a few hours.

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Sources of Biocatalysts and Techniques for Biotransformation

A wide variety of biological catalysts can be used for biotransformation reactions. These
include growing cells, resting cells, killed cells, immobilized cells, cell-free extracts,
enzymes and immobilized enzymes. The most important sources of biocatalysts and the
procedures employed for biotransformation are briefly described.

Growing cells

The desired cells are cultivated in a suitable medium. As the growth of the cells occurs (6-
24 hours), a concentrated substrate is added to the culture. Sometimes, addition of
emulsifiers (Tween, organic solvents) is required to solubilize substrates and/or products
e.g. steroid biotransformation. The substrate conversion to product can be monitored by
spectroscopic or chromatographic techniques. Biotransformation can be terminated when
the product formation is optimum.

Non-growing cells

The non-growing cells are preferred for biotransformation reactions for the following
reasons.

a. Very high concentration of substrate can be used (with high substrate

concentration, growing cells stop their growth).
b. Cells can be washed and used and thus there will be no contaminating substances.
c. Conversion efficiency of substrate to product is high.
d. Product isolation and its recovery are easy.

Immobilized cells

Bio-transformations can be carried out continuously by employing immobilized cells.

Further, the same cells can be used again and again. Several bioconversions with single or
multistage reactions are in fact carried out by using immobilized cells e.g. commercial
production of L- alanine and malic acid.

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Immobilized enzymes

Cell-free enzyme systems in the form of immobilized enzymes are most commonly used
in bio-transformations, due to the following advantages.

a. No occurrence of undesirable side reactions.

b. The desired products are not degraded.
c. There is no transport barrier across the cell membrane for the substrate or product.
d. The isolation and recovery of the product is simpler and easier.

Several immobilized enzyme systems have been developed for bio-transformations e.g.
glucose isomerase, penicillin acylase.

Product Recovery in Bio-transformations

In most biotransformation reactions, the desired end products are extracellular. The
product may be either in a soluble or suspended state. When whole cells are used, they
have to be separated and repeatedly washed (with water or organic solvent) as required.
The extracted product can be recovered by employing the commonly used techniques-
precipitation by salts, extraction with solvents, adsorption to ion-exchangers etc. The
volatile products can be recovered by direct distillation from the medium.

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