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  inflamationat
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 acid-
rich medium
composition in an
open aeration
incubation vessel
maintained at about
32 - 39C
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