Molecular farming is a term coined to describe the application of molecular-biological techniques to the synthesis of commercial products in plants.

A wide range of products have already been identified as likely targets for molecular farming , including a variety of carbohydrates, fats, and proteins.

Plants produce a range of commercially valuable carbohydrates. The two most abundant carbohydrates are:

Cellulose

Starch

There are a number of other carbohydrates that it could be attractive to produce in transgenic plants. Including oligofructans and cyclodextrins.

The major starch producing crops cereals and potatoes- are already widely grown to produce starch as a chemical feedstock.

Granulle- bound starch synthase(GBSS) for synthetis of amylose

Starch synthesis involves two major classes of enzymes:

Starch-branching enzymes(SBEs) which create branches in the starch molecule

Starch comprises amylose and amylopectin, so effectively there is a branch in the biosynthetic pathway leading to one or the other form of starch.

The proportion of amylose/amylopectin is normally about

The phenotype of waxy mutants of maize is characterized by a starch with no amylose that gelatinizes easily

The waxy starches have been produced in transgenic potatoes by antisense inhibition of GBSS1.

high-amylose starches with limited branching have high gelling properties, which can be used for confectionary

The potential exists for carrying out further biotransoformations on the starch in the plant, rather than by chemical or fermentation processes after extraction.

A bacterial cyclodextrin glycosytransferase gene from Klebsiella pneumoniae was fused to a plastid targeting sequence and placed under the control of the promoter from the patatin gene.

Another example of a carbonhydrate targeted for production in transgenic plants is the polyfructans. These compounds are soluble polymers of fructose that are synthesized and stored in the vacuole.

inulins are the major storage carbohydrate found in storage roots such as Jerusalem artichocke
levans are widespread in the leaves and stems of grasses, including major cereal crops such as wheat.

The biosynthetic pathway of fructans in plants is a twostage process but in certain bacteria, such as Bacillus subtilis, very high-molecular-mass levans are produced by a single reaction The sacB gene of Bacillus Subtilis or Bacillus amyloliquifaciens, which encodes a levansucrase, was transformed into maize, tobacco, potato, and sugar beet.

35S

35S

AIMV

CPY

sacB

3nos

In maize, a Bacillus amyloquifacens sacB gene was expressed in the endosperm under the control of a zein promoter

Two different vacuolar targeting sequences:

the sweet potato sporamin signal peptide and vacuolar targeting sequence were fused to the N-terminal of enzyme the barley lectin vacuolar targeting sequence was fused to the C-terminal end of the enzyme

Plant genes econdig inulin biosynthetic enzimes have also been used for genetic manipulation. Short oligofructans have been produced in sugar beet using a gene econding the 1-SST enzyme from Jerusalem artichocke.

This transgenic sugar beet has therefore been renamed the fructan beet.

The fructan beet has direct applications in the nutraceuticals market. Short-chain fructans are almost as sweet as sucrose, and can be used as substitute sugars in foods and drinks. Fructans are not digested in the gut, and can therefore be marketed as a low-calorie sweetener.

The Production of fructans in transgenic plants has been critically reviewed by Cairns(2003)

Sucrose

Sucrose

His calculations indicate that although significant amounts of fructan may accumulate in transgenic plants, the rate of synthesis represents a miniscule fraction of the primary carbohydrate flux

Sucrose

Polyfructans

Lipids are already produced in large quantities from major crops such as oilseed rape, soybean, and maize, for industrial as well as food purpose.

The oils produced by the major oil crops of the world consist mainly of palmitic, stearic, oleic, linoleic and linolenic acids.

The California bay tree contains a very high proportion of lauric acid in its seeds, and an acyl-ACP thioesterase that specifically hydrolyses lauroyl-ACP has been clone from this source.
The introduction of this gene into oilseed rape causes a high proportion of lauric acid to accumulate in the seed oil. Most importantly, field test show that these plants grow normally and produce normal yields.

Erucic acid is one of the constituents of brassica oils, however, although erucic acid is valuable as an industrial oleochemical, it is nutritionally unsuitable for human consumption.

Transformation of oilseed rape with a combination of -ketoacyl-Coa synthase and 22:1 acyl- Coa:lysophosphatidic acid acyltransferase produced 60% erucic acid.

There are 210 known types of fatty acid produced in plants, but most of these are not found in the major crop plants and would be difficult to produce commercially in their host plants.

Certain polyunsatured fatty acids have pharmaceutical or nutraceutical value.

-linolenic acid and arachidonic acid, which are essential fatty acids for humans and precursors of eicosanoids.

The synthesis of these compounds involves a specific subclass of microsomal desaturates called front-end desaturates

One of the most imaginative examples of molecular farming has been the attempt to produce biodegradable plastics in plants. These compounds are currently produced by microbial fermentation, but a number of experimental studies have been carried out to determine the feasibility of producing them In bulk in plants.

Polyhydroxybutrate(PHB) is the bestcharacterized polyhydroxyalkanoates (PHAs)

The genes for the three enzymes involved in the pathway (successively the phaA, phaB, and phaC genes) have been cloned from Alcaligenes eutrophus.

All three genes were transformed into Arabidopsis, and targeted to the chloroplast.
In the first generation of experiments, each gene was separately fused to a sequence encoding the transit peptide plus N-terminal fragment of the Rubisco small subunit protein (rbcS), and expression of each construct was directed by the cauliflower mosaic virus 35S promoter

the bioplastics accumulatd as 0.2-0.7 m granules in the plastids, to levels of up to 14% of plant dry weight, and there was no observable effect on growth or fertility.

A team from Monsanto used the three genes for the PHB biosynthetic pathway from the bacterium Ralstonia eutropha and fused each one to a seed-specifc promoter.

These were transferred into a single, multigene vector, which was used to transform oilseed rape

PHAs are not the only type of bioplastic that could be produced in plants. Amino acid polymers are found in certain bacteria and are already exploited for various commercial purposes.