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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 • • • • • • • Amylase Protease Lipase Cellulase Pectinase Phytase ….. Factor affecting their affectivness -Temperature - pH Application of Enzyme • • • • • 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 • • • • Very strong compare with steel Need method to produce artificially Spider is cannibal Application could be – – – – – – – 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?