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Single cell oil production

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  • 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. 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. BIOSYNTHESIS OF OIL IN MICROBESGLYCOLISIS
  • 4. GLYCOLISIS CONTINUED ………..
  • 5. PRODUCTION OF ACETYL COA
  • 6. FATE OF ACETYL CoA ACETYL CoALIPOGENESIS CITRIC ACID CYCLE
  • 7. CONVERSION OF ACETYL CoA TO MALONYL CoA
  • 8. MALONYL CoA TO FATTY ACYL COA
  • 9. FATTY ACID TO FATTY ACYL COA
  • 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. 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. 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. 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. Vector : pINA1317USES A STRONGRECOMBINANTGROWTH PHASEDEPENDETPROMOTER,hp4d.
  • 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. CLONING VECTOR PMD19-T-INU1CARRIES INULASE GENECLONED FROMK.maxianus CBS 6556
  • 17. CONSTRUCTION OF RECOMBINANT EXPRESSION VECTOR
  • 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.  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. TIME FACTOR
  • 21. EFFECT OF SUGAR
  • 22. EFFECT OF NITROGEN
  • 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. FATTY ACID % PERCENTAGEC14:0 1.68C16:0 28.32C16:1 2.12C18:0 4.65C18:1 58.48C18:2 2.14
  • 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. 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.  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