Single cell oil production

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Single cell oil production

  1. 1. WHAT IS THAT EXACTLY ???Single cell oils (SCO) are the edibleoils extracted from micro-organisms-the single-celled entities that are atthe bottom of the food chain. Singlecelled entities are algae, bacteria,yeast . Lipids are produced by theorganism from basic units likecarbohydrates for their cellmetabolism & survival.
  2. 2. WHY IT IS SO SPECIAL TO OIL TECHNOLOGIST ( LIKE ME ) ??? THEY CAN BE SOURCE OF OILS & FATSWHICH CAN SERVE AS RAW MATERIALFOR OLEOCHEMICALS INDUSTRIES THEY CAN BE SERVE AS MODIFIEDEDIBLE OILS THEYARE SOURCES OF PUFA ANDOTHER ESSENTIAL FATTY ACID. THEY CAN BE USED TO PRODUCEMODERN GENERATION BIOFUEL
  3. 3. BIOSYNTHESIS OF OIL IN MICROBESGLYCOLISIS
  4. 4. GLYCOLISIS CONTINUED ………..
  5. 5. PRODUCTION OF ACETYL COA
  6. 6. FATE OF ACETYL CoA ACETYL CoALIPOGENESIS CITRIC ACID CYCLE
  7. 7. CONVERSION OF ACETYL CoA TO MALONYL CoA
  8. 8. MALONYL CoA TO FATTY ACYL COA
  9. 9. FATTY ACID TO FATTY ACYL COA
  10. 10. Glycerol ATP phosphatase Dihydroxyacetone phosphate glycerol fatty acyl CoA kinase ADP Glycerol-3-P CoA fatty acyl CoA Acyldihydroxyacetone phosphate NADPH CoA Lysophosphatidic NADP+ acid fatty acyl CoA Pi CoA Phosphatidic Diacylglycerol acid fatty acyl CoAFORMATION OF OIL CoA Triacylglycerol Figure 3. Formation of phosphatidic acid from glycerol-3-P or DHAP, and its conversion to triacylglycerol
  11. 11. WE CAN CHANGE THE ACTIVITY OF ENZYMEIN PATHWAY TO PRODUCE LIPID. WE CAN IMPROVE THE YIELD OF OIL BYMAKING APPROPRIATE CHANGE INMICROORGANISM . WE CAN FORCE MICROBES TO PRODUCEOIL FROM THOSE ORGANIC MATTER WHICH ISNOT USEFUL TO US. TO EXPLOIT NEWER TACTICS TO ACHIEVETHE SAME.
  12. 12. SO MY CURRENT DISCUSSIO N WILL BE BASED ON RESEARCHWORK CONDUCTED BY CHUN-HAI ZHAO & WEI CUI AT UNESCOCHINESE CENTER OF MARINE BIOTECHNOLOGY AND INSTITUTEOF MARINE BIODIVERSITY AND EVOLUTION, CHINA.THE PAPER FOCUS ON HOW RDTHELPED IN IMPROVING OILPRODUCTION FROM YEAST BYEXPRESSION OF INULASE GENE INTOIT.
  13. 13. PurposeTo express inulase gene in oleogeinous yeast yarrowcialipolytica and thus obtain oil from inuln containing material. Host : yarrowia lipolytica Reason : 1) fast growth rates 2) high oil content 3) Resemblance of their triacyglycerol in plants.
  14. 14. Vector : pINA1317USES A STRONGRECOMBINANTGROWTH PHASEDEPENDETPROMOTER,hp4d.
  15. 15. INULIN PRESENT AS RESERVECARBOHYDRATE IN ROOTS &TUBERS OF PLANT LOW COST MATERIAL WAS NOT USED BECAUSEBECOZ OLEOGINOUSMAICROBES WERE NOT ABLETO SECRETE INULASE. SO EXPRESSION OF INULASEGENE PROMOTES SECRETIONOF INULINASE AND THISSUGAR MOIETY WASCONVERTED TO OILS.
  16. 16. CLONING VECTOR PMD19-T-INU1CARRIES INULASE GENECLONED FROMK.maxianus CBS 6556
  17. 17. CONSTRUCTION OF RECOMBINANT EXPRESSION VECTOR
  18. 18. CLONING VECTOR WAS PREPARED BYSAMBROOK ET AL.Y LIPOLYTICA WAS TRANSFORMED BYXUAN.ET.ALTO AMPLIFY INU1 GENE BY PCR,FORWARD PRIMER WAS PPU & REVERSEPRIMER WAS PPD. CONFIRMATION OF INTEGRATION OFINULASE GENE WAS DONE BY PCRFOLLOWED BY CHECKING ATAUTOMATED GEL DOCUMENTATION &ANALYSIS SYSTEM.
  19. 19.  DETERMINATION OF INULASE ACTIVITY WASDETERMINED ONYERMS OF INULASE UNIT ( U)WHICH IS AMOUNT OF ENZYME THAT PRODUCES 1MICROMOLE OF REDUCING SUGAR PER MINUTEUNDER THE ASSAY CONDITION. EXPERIMENT WAS CONDUCTED IN 250 mLCONICAL FLASK CONTAINIG 50 ml OF SINGLECELL OIL PRODUCTION MEDIUM & 5 ml CELLMEDIA. LIPIDS WAS EXTRACTED BY SOLVENTEXTRACTION .IT WAS REFINEDIT WAS CONVERTED INTO FAME & GC ANALYSISWAS PERFORMED TO CHECK FATTY ACIDCOMPOSITION
  20. 20. TIME FACTOR
  21. 21. EFFECT OF SUGAR
  22. 22. EFFECT OF NITROGEN
  23. 23. IN THE NUTSHELL……. TRANSFORMANT Z 31 ACCUMULATED 46.3 % (WT/WT) OF OIL IN ITS CELL AND CELL DRY WEIGHT WAS 11.6 g PER LITRE& ABOUT 10.7% OF THE SUGAR WAS CONVERTED INTO ITS CELL OIL.
  24. 24. FATTY ACID % PERCENTAGEC14:0 1.68C16:0 28.32C16:1 2.12C18:0 4.65C18:1 58.48C18:2 2.14
  25. 25. SCOPE THE MENTIONED EXPERIMENT WAS TERMED A“ HIT “ AT AOCS 2010.RESEARCH IS NOW BEING DONE TOIMPROVE FURTHER PRODUCTION BY OVEREXPRESSION OF GENE ENCODINGATP CITRATELYASE, FATTY ACID SYNTHASE AND ACYL COACARBOXYLASE AND DISRUPTION OF GENESENCODING GLYCEROL-3-PHOSPHATEDEHYDROGENASE AND ACYL CoENZYME AOXIDASE
  26. 26. CONCLUSION SINGLE CELL PRODUCTION IS NOT COMMERCIALIZED METHOD. PROCESS CAN BE COMMERCIALIZED BY1) RESEARCH & DEVELOPMENT IN THIS FIELD .2) INVESTMENTS BY BIG CORPORATES.3) PROMOTION OF THS TECHNOLOGY AT VARIOUS CONFERENCES WORLDWIDE. & MOST IMPORTANT
  27. 27.  WWW.GOOGLE.COMWWW.WIKIPEDIA.ORGPRINCIPLE OF GENE MANIPULATION BYPRIMROSABIOCHEMISTRY BY LEHNINGER.MODERN TRENDS IN OILS & FATSTECHNOLOGY BY OP NARULA RESEARCH PAPERS ONWWW.SCIENCEDIRECT.COM

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