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Protein as Biotechnology Products
By
Dr. Thanut Amatayakul
Biotechnology Division
Faculty of Agricultural Product Innovation and
Technology
Srinakharinwirot University
Outline
• Some basics and overview of Proteins
• Protein as Biotechnology Products
Structure of Protein
http://www.umass.edu/molvis/workshop/prot1234.htm
Protein synthesis
• Start from DNA
• Involve transcription (from DNA to mRNA) and
from mRNA to protein
• Post protein synthesis
• Occur in Nucleus and Cytoplasm (Eukaryote) or
Cytoplasm (Prokaryote)
• More information can be found in
http://www.nature.com/scitable/topicpage/ribos
omes-transcription-and-translation-14120660
Video protein synthesis
http://www.youtube.com/watch?v=5iS4CRPPDus
Genetic engineering
Nisin
ที่มา: http://commons.wikimedia.org/wiki/File:Nisin_2d_amino_structure.JPG
• An antimicrobial agent
• Produced by Lactococcus lactis
• Used as a food preservative at low concentration (up to 250 ppm) in
heat processing / low pH foods
• GRAS (Generally regard as safe)
Mechanisms
Marian Petre,,
Advances in Applied
Biotechnology, ISBN
978-953-307-820-5,
2012
Nisin
• Effective against Gram positive bacteria only
• Bind to lipid II blocking peptidoglycan
formation
• Creating pore/ leakage
• Cell death
Production
Nisin production in batch fermentation with the ISC of 40 g l−1. Sucrose
concentration (filled square), lactic acid (open square), nisin titer (circle), CDW
(triangle)
Applied Biochemistry and BiotechnologyPart A: Enzyme Engineering and Biotechnology, Wu & Wang, 2008, Variable Volume Fed-Batch Fermentation for Nisin Production by Lactococcus lactissubsp. lactis W28
Enzyme
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Amylase
Protease
Lipase
Cellulase
Pectinase
Phytase
…..
Factor affecting their affectivness
-Temperature
- pH
Application of Enzyme
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Food and Beverages
Textiles
Paper
Environment
Detergent
– Please give some examples for the above topic
• What kind of Enzyme?
• It’s Roles for that application?
Enzyme Production
http://www.youtube.com/watch?v=T8mOMMmzS18
Insulin
• It is a hormone produced from b-cells of the
pancreas
• It stimulates absorption of glucose into the
cells of liver, skeleton muscle, fat tissue
• Insluin: 51 amino acids, and has a molecular
weight of 5808 Da. It is a dimer of an A-chain
and a B-chain, which are linked together
by disulfide bonds.
• Serious problem for Diabetes mellitus
Diabetes
• the person has high blood glucose (blood
sugar), either because insulin production is
inadequate (type 1), or because the body's
cells do not respond properly to insulin (type
2), or both.
http://commons.wikimedia.org/wiki/File%3AMain_symptoms_of_diabetes.png
Insulin
Video Insulin
http://www.youtube.com/watch?v=MJ_6oXaLRj4
ACE inhibitory peptide
• Short peptides
• products of protein digestion
• Some peptides have angiotensin converting
enzyme inhibitory properties
• Block ACE
• Prevent hypertension (high blood pressure)
ACE inhibitory peptide
• It can be made by enzymatic hydrolysis of
proteins
Origin
Sequence/name
IC50
(µmol/L)
Subjects
Reference
Milk (b-casein)
VPP, IPP
9.0, 5.0
SHR, Human
Nakamura et al.,
1995; Seppo et al.,
2003
Milk
(β-lactoglobulin)
IPA (β-lactosin A)
141.0
ALPM (β-lactosin B) 928.0
SHR
Abubakar et al.,
1998; Murakami et
al., 2004
Fish
(sardine muscle)
VY
26.0
SHR
humans
Kawasaki et al.,
2000; Matsufuji et
al., 1995
Fish
(bonito muscle)
LKPNM
LKP
2.4
0.32
SHR
Fujita et al., 1999
Effect of various enzymes on ACE inhibition during prolong digestion
Unpublished result from research of Dr. Thanut Amatayakul
Spider Silk
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Very strong compare with steel
Need method to produce artificially
Spider is cannibal
Application could be
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Bullet-proof clothing
Wear-resistant lightweight clothing
Ropes, nets, seat belts, parachutes
Rust-free panels on motor vehicles or boats
Biodegradable bottles
Bandages, surgical thread
Artificial tendons or ligaments, supports for weak blood
vessels.
http://www.laboratory-journal.com/science/material-science/artificial-spider-silk
Question
• Structually, What is the differences in protein
synthesis between prokaryote and eukaryote?
• From the lecture, please conclude about the
method of making protein as biotechnology
products
• What is protein engineering?