Synthesis of Recombinant

Human Insulin

Data of Diabetic Patient

Since Banting & Best invented insulin hormone in

1921, amount of diabetic patients increased
worldwide.
Their glucose level increased due to disorder of
insulin production treatment by insulin injection
from animal pancreas
Insulin is produced and secreted by beta cells of
Langerhans pancreas, that regulates the use and
storage of carbohydrates.

Problem?

Animal insulin swine and cow

Human insulin slight different in composition
As a consequence : human body produces
antibodies against the animal insulin inflamation
at the area of injection
Long-term complication occurred
Decreasing of animal insulin production

Structure of Insulin
Chemically, insulin is a small and simple protein , consists of
51 amino acids, 30 construct 1 polypeptide chain and 21
amino acids construct the second chains. Both chains are
linked by disulfide bond

Synthesis of human insulin using E.coli

Human Insulin is produced by

inserting gene of insulin at
suitable vector of Escherichia coli
(production of Humulin, Eli Lily) by
recombinant DNA Technology

Selectively
obtaining insulin
producing beta
epithelioid cells
from a pancreas, a
small circular DNA
in a bacteria, called
plasmid, modified
by inserting human
DNA gene for a
precursor proinsulin

Plasmid containing proinsulin gene is

then transformed into Eschericia coli

Dispersing cells in a
nutrient amino acidrich medium
composition in an
open aeration
incubation vessel
maintained at about
32 - 39C

Protein formed is partly B-galactosidase, on chain A and chain B of the

insulin A and B are extracted from fragments of B-galactosidase and
purified

Separating the
cells from the
media,inactive
bacteria by heat
sterilization, and
extracting
genomic material
from the cells,

Proinsulin harvested
by cutting by
restriction enzyme
to produce human
insulin

Further Process
is centrifugation
and eliminating
unused cells.
Purification is
carried out by
liquid
chromatography
and crystallization