Putrescine Lyndsay Grover Biotechnology Technician - Health CHEM April 4 th 2011

Transcription

1 Putrescine Lyndsay Grover Biotechnology Technician - Health CHEM April 4 th 2011

2 Introduction Putrescine is found in living and dead organisms due to the breakdown of amino acids and is the main culprit for the foul odour in putrefying flesh, much like the related compound cadaverine. They are related to the compounds spermine and spermidine and all together they are included in the polyamines family; putrescine being one of the simplest compounds existent in the family. Polyamines play various biological functions and exist in all living things. The have been shown to be crucial in the binding of nitrogenous bases in DNA synthesis. The reasoning for my choice of this compound was the nature of it, it being such a simple compound but being involved in such important biological processes. The main purpose of it is such a simple role; the putrefaction of animal matter; that one can overlook the role it plays in the biological function of all human beings. Especially that involving DNA synthesis, the idea that one compound is involving both in the building blocks of life as well the process of destroying it once it has expired is astounding the multi-purpose nature of such a simple compound. As well an interesting factor in my decision of this compound involves the current scientific research surrounding it and the polyamines. Currently scientists are looking into connections between cell growth and the concentration of polyamines and how it affects cancer cells and the growth of tumors. Not only is a great breakthrough in terms of medical science to discover new methods to fight cancer and to potentially reduce the destruction it has on the human body considerably. It is also an interesting fact to observe the notion that a compound that is involved in death and decay is being used to potentially prolong lives and even save people from dying of an illness altogether. It is involved in something greater than its normal biological function. Technical Information I. Name and Formula: Putrescine is the industrial name for the compound, which is also known to be spelled as putrescin. The IUPAC name for it is butane-1,4-diamine or butanediamine as well as 1-4, diaminobutane. The chemical formula for Putrescine is NH 2 (CH 2 ) 4 NH 2 and the molecular formula is C 4 H 12 N 2. II. Physical Properties: Putrescine is a foul smelling compound and at room temperature it is a clear colourless liquid. It has a boiling point range of C, a melting point of 26 C and a flash point at 64 C. It is freely soluble in water and not known to be soluble in any other liquid. It is highly toxic in large doses, although in rats it had a low acute oral toxicity, with the dosage only being 2000mg/kg of body weight. It is also known to be a severe eye, respiratory tract and digestive system irritant, as well a regular skin irritant.

3 III. Uses: Putrescine in nature is used for the breakdown of amino acids in animal matters as well it is the contributing factor to bad breath and bacterial vaginosis. It is also of the family of polyamines which are crucial in cell growth and differentiation. Polyamines are responsible for the bonding of nitrogenous bases to the phosphate groups and sugars. Research has been shown linking the surplus of polyamine levels in cells to arrest the processes of cell growth, differentiation and division. Putrescine has many industrial medical applications. For example putrescine is used as a TG inhibitor as a treatment for scar tissue for patients recovering from surgery or suffering serious burns. It also has other various practical purposes; solutions created with putrescine can be used to prolong the life and firmness of fruits and plants. This is used to prevent fruits and plants from decaying while in storage or during transportation. Current research is trying to use this linkage to stop cell growth and prevent division in tumor cells. By modifying the genetic makeup of tumor cells to promote the creation and usage of polyamines or by increasing the polyamine concentration via polyamine producing enzymes researchers hope to prevent tumor growth. This also means they would be able to prevent tumors from spreading and also constantly evolving to be unaffected by the body s natural immunities. If successful trials of this are conducted researchers may find a way to increase the likelihood of surviving cancer. IV. Sources: Putrescine is found naturally in the decaying flesh of animal matter, but also is found in semen and microalgae. Polyamines, of which putrescine is one of the simplest, occur in both eukaryotic and prokaryotic cells as it has purposes for cell growth and differentiation. It also has the capable of being produced chemically from succinonitrile and biosynthetically from Escherichia coli. V. Production: How putrescine is produced naturally in eukaryotic cells is the base molecule of arginine is only converted to putrescine via the ornithine decarboxylase enzyme. Where as in prokaryotes and plant cell they have the option to take an alternative route of synthesis where arginine is acted upon by arginase decarboxylase and then agmatine ureohydrolase to form putrescine.

4 Source: ( Putrescine can be synthesized chemically through the hydrogenation of succinonitrile, which occurs when hydrogen cyanide is added to acrylonitrile. Putrescine can also be produced through biosynthesis in a process which involves an engineered strand of Escherichia coli which produces putrescine in high concentrations of glucose mineral salts. In this process the putrescine degradation and utilization pathways are inactivated to create a base chain. Next the ornithine carbalymoyltransferase chain I gene is deleted to allow for more available precursors available for putrescine synthesis. After a suitable base chain is created ornithine decarboxylase is amplified via plasmid recombinant DNA technology to increase the output of putrescine. More modifications are made to the enzyme to overexpress the gene for ornithine production in order to create a greater ornithine pool. As well the stress response RNA polymerase sigma factor (RpoS) is deleted to prevent the ornithine decarboxylase system from shutting down when producing more than the normal amount of putrescine. All the modifications allow for a drastic increase in the output of putrescine, making it possible to create large batches for industrial, medical and commercial usage. Once putrescine is created in a chemical or biotechnological setting it is packaged and sold to individual laboratories that request samples of it for their research. VI. Fate: Putrescine degrades after it has served its biological function, but unlike most compounds that degrade in one certain fashion, putrescine degrades in 5

5 variants, all different processes involving various reactions that all result in the created of 4-aminobutyrate. Putrescine degradation variant I: In this pathway putrescine is transformed with 2-oxoglutarate to produce L-glutamate and 4-aminobutanal. Those two molecules are then oxidized to create 4-aminobutyrate. Source: ( PWY) Putrescine degradation variant II: In this pathway with the usage of one ATP putrescine it γ-glutamylated to produce γ-glutamyl-l-putrescine. That resulting compound is oxidized to create γ-glutamyl-γ-aminobutyraldehyde and is then dehydrogenated 4-(glutamylamino) butanoate. The last step in the process requires the removal of the γ-glutamyl group via hydrolysis to create 4- aminobutyrate. Source: ( Putrescine degradation variant III: In this pathway putrescine goes through acetylation and oxidative deamination via 4-acetamidobutanal to create N- acetyl-4-aminobutyrate which is then transformed into 4-aminobutyrate. Source: ( Putrescine degradation variant IV: In this pathway putrescine is acted upon by a putrescine oxidase enzyme. This reaction creates 4-aminobutanal,

6 which can create either 4-aminobutyrate or 1-pyrroline. This process is spontaneous and is unknown which compound will be the final product. Source: ( Putrescine degradation variant V: This pathway is identical to putrescine degradation variant I, the only exception is that in this version instead of using 2- oxoglutarate this variant using pyruvate as its preferred acceptor. Source: (

7 Model

8 Reference Abraham KA (1968). "Studies on DNA-dependent RNA polymerase from Escherichia coli. 1. The mechanism of polyamine induced stimulation of enzyme activity." Eur J Biochem 5(1); Heby, Olle Putrescine, Spermidine and Spermine, NIPS, Vol:1, pg:12-15, 1986 MetaCyc Putrescine Degradation, [ type=pathway&object=pwy-3], Mar 30, 2011/Apr 1, 2011 MSDS (Putrescine), [ Jun 22, 2004/Apr 1, 2011 Putrescine [ Mar 28, 2011/Mar 15, 2011 Rhodes, David, Putrescine Synthesis - Polyamines, nonprotein amino acids and alkaloids [ Oct 1, 2009/Mar 15, 2011 Zhi-Gang Qian, Xiao-Xia Xia, Sang Yup Lee, Metabolic Engineering of Escherichia coli for the Production of Putrescine: A Four Carbon Diamine, Biotechnology and Bioengineering, Vol: 104, pg: , 2009