Downloaded 500 times
Uploaded byNeha Patel
Plant metabolite engineering
This document discusses plant metabolic engineering. It begins with an introduction to metabolic engineering and explains that it involves modifying biochemical reactions or introducing new ones using recombinant DNA to direct the formation of products or modify cellular properties. It then discusses plants as natural factories for metabolic engineering due to their low cost and ease of growth. Various strategies for plant metabolic engineering are covered, including modifying rate limiting steps or diverting flux. Applications like producing antibodies, edible vaccines, polymers and pharmaceuticals in plants are also summarized. Emerging technologies and the future of the field are then outlined.
More Related Content
Similar to Plant metabolite engineering
Plant metabolite engineering
- 1. Plant Metabolic Engineering PLANTMETABOLIC ENGINEERING BY NEHA P PATEL M.Sc II
- 2. Outline of theTalk Introduction to Metabolic Engineering Plants as natural factories Metabolic Engineering in Plant Choice of plant system for ME Stratigies for ME Applications Emerging technologies Future
- 3. Introduction to MetabolicEngineering It is the directed improvement of product formation or cellular properties through the modification of specific biochemical reaction(s) or the introduction of new one(s) with the use of recombinant DNA technology
- 4. METABOLIC ENGINEERING Metabolic Networks MODIFICATION recombinant DNA technology ANALYSIS FluxQuantification Analysis of Flux Control Cell improvement
- 5. OTHER TERMS FORME 1.MOLECULAR BREEDING 2.PATHWAY ENGINEERING 3.CELLULAR ENGINEERING
- 6.
- 7. What’s so hardabout modeling plants? Plant cell metabolism is complex… Collectively, plants produce over (primary and secondary) metabolites less information is available as compared to bacteria ProteinsGenesBase pairs 1366 (30%+)4000-50004.6ME.coli 3500 (10%+)30000135MArabidopsis
- 8. WHY PLANTS ASNATURAL FACTORIES Cheap availability easy to grow Low cost of growing plants Provide eukaryotic system Plants cells are highly compartmentalized
- 9. PLANT METABOLIC ENGINEERING Plant metabolic engineering involves the manipulation of existing metabolic pathways by either increasing or diverting flux to desired or from undesired products, respectively, or the generation of chemical entities not normally found in the plant production system (cells or whole plants),through the introduction of genes from other organisms.
- 10. Essential elements inthe toolbox of the metabolic engineer are mechanisms to eliminate or over express gene activity Introduction of new pathway
- 11. Strategies available toeliminate the activity of specific enzymes in a pathway involve one of several possible approaches Identification of a mutant gene for the corresponding enzyme. Knocking out gene function by targeted RNA degradation. Interfering with protein function using specific inhibitors or antibodies.
- 12. Process of RNAinterference
- 13. The expression ofgenes in plants or plant cells is dependent on several factors that include: method to introduce genes into the plant Promoters to direct gene expression in the appropriate spatial and temporal landscape. source for the gene encoding the enzyme of interest.
- 14. CHOICE OF PLANTSYSTEM FOR METABOLIC ENGINEERING Whole Plants Plant Cells in Culture
- 15. STRATEGIES FOR INCREASINGFLUX OF EXISTING PATHWAYS Manipulating the Activity of ‘‘Rate Limiting’’ Steps Metabolic Flux Analysis and Modeling Expression of Multiple Genes in Plants:Progress and Limitations Diverting Flux Using Loss of Function Approaches Targeting Entire Pathways withTranscription Factors
- 16. PLANTS AS BIOREACTOR Antibodies ediblevaccines Polymers Pharmaceutical drugs etc
- 17. ANTIBODIES Stable integration ofgene as compare to microorganisms Provide eukaryotic system for production Low cost production as compare to fermentation-it costs approx. $5,000 /gm to produce antibodies from hybridoma technology cells in culture,$10 to 100 /gm to produce antibodies from transgenic plants. Foreign proteins are mostly produced in seed But often not properly glycosylated
- 19. PRODUCTION OF EDIBLEVACCINE
- 20.
- 21. Transgenic Plant Vaccines– Potential for Edible Vaccines Pros No need for equipment such as needles No personnel would required Ability for mass immunization at low cost Microorganisms could produce them in foods such as yogurt and cheeses but would require processing, adding cost and limiting accessibility Plants offer a nutritious vector, especially useful in third world countries where they would be very important
- 22. Cons Restricted to plantswhose products are consumed raw to avoid degradation during cooking Concerns that ingested proteins would be broken down in the gut by proteinases Trying to prove that antigens being encapsulated by plant tissue will resist immediate breakdown Accidental consumption due to human error in storage and transportation of crops Low amounts of highly immunogenic antigen caused no detectable side effects in mice however much more work must be done in researching different doses, mixes and antigenicities before this can be removed as a possible threat In an experiment with an edible rice vaccination immunizing against cholera they were proven effective after being stored at room temperature for 1.5 years
- 23. Polymers polyhydroxyalkanoates (PHA) inplants. polyesters of 3-hydroxyacids have unique biodegradable and elastomeric properties used in medical industry, and for making environmental friendly plastics PHB granules in Arabidopsis mesophyll cell nucleus
- 24. (PHB/PHV block copolymer) Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Biopolymer production GlycerolPropionate Acetyl-CoAPropionyl-CoA Acetoacetyl-CoA 3-Ketovaleryl-CoA 3-Hydroxybutyryl-CoA3-Hydroxyvalery-CoA Acetyl-CoA HSCoA 3-Ketothiolase (PhaA) NADPH NADP+ Acetoacetyl-CoA Reductase (PhaB) P(HB-co-HV) HSCoAHSCoA PHA Synthase (PhaC)
- 25. resulting in asignificant accumulation of polymer. genes from bacteria were successfully introduced into the plastids of Arabidopsis thaliana, Bacterial such as Alcaligenes eutrophus, PHA is synthesized from acetyl coenzyme A in three steps catalyzed by three enzymes whose genes are organized on a single operon.
- 26. Plants as Pharmaceutical Factories Haveastonishing potential for the biosynthesis of small (<1000 Da) molecules. Many Secondary metabolites as drugs.example- morphines,steroids. Many phytochemicals like flavonoids,anthocynins, ,phenolic compounds etc
- 27. Figure showing Drugproduction in plants
- 28. Using Plant Compartmentsfor Chemical Sequestration PROBLEM-Massive accumulation of metabolites if transgenes are expressed constitutively throughout the plant SOLUTION-1.the use of tissue-specific promoters, allowing for accumulation in either a specific organ or tissue. SOLUTION-2.gene expression can be controlled in such a way that biosynthetic enzymes or metabolites are directed to specific cell compartments such as the vacuole or chloroplast pathway intermediates or final products could be sequestered in specific subcellular compartments
- 29. EMERGING TECHNOLOGIES Plant Diversityas a Source of New Genes-unique biosynthetic pathways that offer opportunities to use genomic approaches to capture this biochemical diversity and provide tools for manipulating plants and other organisms that do not produce the compounds or where accumulation is blocked at a particular biosynthetic step Gene Shuffling and Directed Enzyme Evolution- Gene shuffling technologies can produce a large number of enzyme variants, by shuffling fragments from an existing library
- 30. The FUTURE Focuson secondary metabolism Progress in ‘omics technologies’. Better use of what we know! Choose model systems we can experimentally validate Apply known constraints Define appropriate objective functions Integrate regulatory mechanisms
- 31.
- 32.
Editor's Notes
- #9 PLANTS CELLS ARE HIGHLY COMPARTMENTALIZED ,FACILITATING ,FOR EXAMPLE ,THE SEPARATION OF SUBSTRATE FROM THE PRODUCTS.