Presented by: sanchari maityM.Sc biotechnology 1st semBioengineering:

custom microbes

Contents what is bioengineering?

domains of bioengineering.Genetic and biomedical engineering.

• Recombinant insulin• recombinant human growth hormone

• Recombinant interferon• Hepatitis B vaccine production

• Phage therapy

Use in bioremediation.

• Superbug• Bio-luminescence while pollutant

degradation

Use in bioprocess engineering.

• various types of expression vectors

What is bioengineering?British scientist and broadcaster Heinz Wolff termed bioengineering in 1954.

Any area of biology mixed with any area of engineering in any proportion.

Principles of biolog

y

Tools of

engineering

Bioengineerin

gUseabl

e,tangibl

e,viable

pdt

engineering

Analytical & synthetic

Cost,practical application

Maths,chemistry,physics,computer

scienceStructure,process,

manufacture

biology

Genome knowledge

Molecular biology

Recombinant DNA technologyMedical,

Research,

genetic engineering

Cellular engineering

Bioprocess engineering

Biomedical engineering

Biomimetics

bioinformatics

Biomedical engineering

• Artificial organs• Replacement joints• Artificial skin.

• Vaccine,drug,hormone production.

Bioinformatics = computer science + biomedicine Discover genetic basis for disease (cancer, diabetes)Develop new diagnostic devices (cDNA chip) cDNA Array

Cellular engineering

bioimmetics

bioinformatics

Genetic engineering

Genetic engineering in brief

GENETIC AND BIOMEDICALENGINEERING

First attempt to produce insuline through rDNA technology:1970s

Human insulin gene and lac operon promoter inserted into E.coli plasmid

July 1980, 17 diabetic patients were treated with recombinant insulin at Guy’s hospital,london :Great success.

First RDT product administered to human and worked well.

Eri Lilly company received approval to market HUMan insULIN under the trade name HUMULIN

Recombinant insulin production

•Insulin A and B chain inserted differently in different E.coli culture

•Selective marker:ampicilin resistant gene

•Do pure culture

•Extract insulin A & B chain seperatelyAnd Mix them.

•Functional insuline

Recombinant hGH production

•Glycosylated cytokines.•2 types exist

Type I (IFN-,and IFN-)Type II (IFN-)

•Cells producing IFNsPlasmacytoid DCs (major producers of IFN- and IFN- )Fibroblasts and epithelial cellsMacrophages and Th1 Cells (predominantly IFN- )

•Stimulates the action of natural killer cell•Several interferon together in a single gene construct: more efficient interferon.

Recombinant human interferon

Select desired interferon genes.

cDNA from specific interferons.

Insert into vector

Vector introduced into host cell( E. coli/yeast cell)

Isolate recombinant interferon from culture medium

Hybrid interferon

Why Yeast vector? yeast suitable:glycosylation of protein similar to mammalian cell.Yield is several fold higher than E. coliHybrid IFN : more reactive in function

THERAPUTIC APPLICATION IFN-α,-β,-ɣ approved for theraputic use in 1986,1993 & 1990A swiss biotechnology firm first marketed IFN- α, named (INTRON)Treatment of large number of viral disease and cancers. Like leukemia,kaposis sarcoma,bladdar cancer,head and neck cancer,renal cell carcinoma,skin cancer,multiple myloma.Other disease:AIDS,multiple sclerosis,genital warts,hepatitis C. common cold,influenza:nasal spray

Recombinant hepatitis-B surface antigen production

Phage therapy

BIOREMEDIATION BY BACTERIA

superbugNew pseudomonas strain developed by Chakrabarty and co-workers in 1970s.

Different plasmids were used and new bacterium was constructed.SUPERBUG.

US patented this SUPERBUG that can degrade a number of hydrocarbons of petrolium simultaniously.

First genetically engineered microorganism patented.

CAM-camphor degradingOCT-octane degradingXYL-xylene degradingNAH-napthalene degrading

Compatible: CAM-OCT plasmid

•Biodegradative pathway gene and lux gene constructWhile degrading pollutants it gives luminiscence.thus we can Conclude that degradation is goin on

The operon of the TOL plasmid pWW0 of Pseudomonas putida encodes a set of enzymes involved in the conversion of toluene and xylenes to their carboxylic acid derivatives. The last gene of the upper operon,

Xylene and toluene(pollutant)

TOL operon of P.putida Carboxilic acid derivative(degraded form)

lux genes is present in marine bacterium, Vibrio fischeri

Biodegradation along with bioluminescence

BIOPROCESS ENGINEERING

p ppa pa

Two vector expression system for producing diametric proteinP-promoter,pa-polyadenylation seq

Recombinant protein can be produced in microorganisms .3 types of expression vectors can be used-1)Two vector expression system2)Two gene expression vector3)Dicistronic expression vector

rh- Growth Hormone, r-Human insulin, Erythropoietin,Follicle stimulating hormone, Interferon, Insulin like growth factor, Tissue Plasminogen Acivator,factor VIII,DNase, Envelope proteing of hepatitis B virus.

Produced by any of these type of method which is convenient

Expression vectors for protein production

Two gene expression vector Dicistronic expression vector

References

6)http://en.wikipedia.org/wiki/Biological_engineering

7)http://www.biopharminternational.com/biopharm/article/articleDetail.jsp?id=3019798)http://www.wiley-vch.de/books/biotech/pdf/v11b_gene.pdf

1)U.Satyanarayana;biotechnology;books and allied (p) ltd,p-144,191,192,197,201

2)http://bioengineering.stanford.edu/

3)http://www.biotecharticles.com/Biotechnology-products-Article/Production-of-Recombinant-Human-Growth-Hormone-Somatotropin-367.html4)www.littletree.com.au/dna.htm

Questions ?....

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