2. WHAT IS THAT EXACTLY ???
Single cell oils (SCO) are the edible
oils extracted from micro-organisms-
the single-celled entities that are at
the bottom of the food chain. Single
celled entities are algae, bacteria,
yeast . Lipids are produced by the
organism from basic units like
carbohydrates for their cell
metabolism & survival.

3. WHY IT IS SO SPECIAL TO
OIL TECHNOLOGIST ( LIKE ME ) ???
 THEY CAN BE SOURCE OF OILS & FATS
WHICH CAN SERVE AS RAW MATERIAL
FOR OLEOCHEMICALS INDUSTRIES
 THEY CAN BE SERVE AS MODIFIED
EDIBLE OILS
 THEYARE SOURCES OF PUFA AND
OTHER ESSENTIAL FATTY ACID.
 THEY CAN BE USED TO PRODUCE
MODERN GENERATION BIOFUEL

5. BIOSYNTHESIS OF OIL IN MICROBES
GLYCOLISIS

6. GLYCOLISIS CONTINUED ………..

7. PRODUCTION OF ACETYL COA

8. FATE OF ACETYL CoA
ACETYL
CoA
LIPOGENESIS CITRIC ACID
CYCLE

9. CONVERSION OF ACETYL CoA TO MALONYL CoA

10. MALONYL CoA TO FATTY ACYL COA

11. FATTY ACID TO FATTY ACYL COA

12. 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 CoA
FORMATION OF OIL CoA
Triacylglycerol
Figure 3. Formation of phosphatidic acid from glycerol-3-P or
DHAP, and its conversion to triacylglycerol

13. WE CAN CHANGE THE ACTIVITY OF ENZYME
IN PATHWAY TO PRODUCE LIPID.
WE CAN IMPROVE THE YIELD OF OIL BY
MAKING APPROPRIATE CHANGE IN
MICROORGANISM .
WE CAN FORCE MICROBES TO PRODUCE
OIL FROM THOSE ORGANIC MATTER WHICH IS
NOT USEFUL TO US.
TO EXPLOIT NEWER TACTICS TO ACHIEVE
THE SAME.

14. SO MY CURRENT DISCUSSIO N WILL BE BASED ON RESEARCH
WORK CONDUCTED BY CHUN-HAI ZHAO & WEI CUI AT UNESCO
CHINESE CENTER OF MARINE BIOTECHNOLOGY AND INSTITUTE
OF MARINE BIODIVERSITY AND EVOLUTION, CHINA.
THE PAPER FOCUS ON HOW RDT
HELPED IN IMPROVING OIL
PRODUCTION FROM YEAST BY
EXPRESSION OF INULASE GENE INTO
IT.

16. Purpose
To express inulase gene in oleogeinous yeast yarrowcia
lipolytica 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.

17. Vector : pINA1317
USES A STRONG
RECOMBINANT
GROWTH PHASE
DEPENDET
PROMOTER,hp4d.

18. INULIN
PRESENT AS RESERVE
CARBOHYDRATE IN ROOTS &
TUBERS OF PLANT
LOW COST MATERIAL
WAS NOT USED BECAUSE
BECOZ OLEOGINOUS
MAICROBES WERE NOT ABLE
TO SECRETE INULASE.
SO EXPRESSION OF INULASE
GENE PROMOTES SECRETION
OF INULINASE AND THIS
SUGAR MOIETY WAS
CONVERTED TO OILS.

19. CLONING VECTOR
PMD19-T-INU1
CARRIES INULASE GENE
CLONED FROM
K.maxianus CBS 6556

20. CONSTRUCTION OF RECOMBINANT EXPRESSION VECTOR

21. CLONING VECTOR WAS PREPARED BY
SAMBROOK ET AL.
Y LIPOLYTICA WAS TRANSFORMED BY
XUAN.ET.AL
TO AMPLIFY INU1 GENE BY PCR,
FORWARD PRIMER WAS PPU & REVERSE
PRIMER WAS PPD.
 CONFIRMATION OF INTEGRATION OF
INULASE GENE WAS DONE BY PCR
FOLLOWED BY CHECKING AT
AUTOMATED GEL DOCUMENTATION &
ANALYSIS SYSTEM.

22.  DETERMINATION OF INULASE ACTIVITY WAS
DETERMINED ONYERMS OF INULASE UNIT ( U)
WHICH IS AMOUNT OF ENZYME THAT PRODUCES 1
MICROMOLE OF REDUCING SUGAR PER MINUTE
UNDER THE ASSAY CONDITION.
 EXPERIMENT WAS CONDUCTED IN 250 mL
CONICAL FLASK CONTAINIG 50 ml OF SINGLE
CELL OIL PRODUCTION MEDIUM & 5 ml CELL
MEDIA.
 LIPIDS WAS EXTRACTED BY SOLVENT
EXTRACTION .
IT WAS REFINED
IT WAS CONVERTED INTO FAME & GC ANALYSIS
WAS PERFORMED TO CHECK FATTY ACID
COMPOSITION

25. TIME FACTOR

26. EFFECT OF SUGAR

27. EFFECT OF NITROGEN

28. 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.

29. FATTY ACID % PERCENTAGE
C14:0 1.68
C16:0 28.32
C16:1 2.12
C18:0 4.65
C18:1 58.48
C18:2 2.14

30. SCOPE
 THE MENTIONED EXPERIMENT WAS TERMED A
“ HIT “ AT AOCS 2010.
RESEARCH IS NOW BEING DONE TO
IMPROVE FURTHER PRODUCTION BY OVER
EXPRESSION OF GENE ENCODINGATP CITRATE
LYASE, FATTY ACID SYNTHASE AND ACYL COA
CARBOXYLASE AND DISRUPTION OF GENES
ENCODING GLYCEROL-3-PHOSPHATE
DEHYDROGENASE AND ACYL CoENZYME A
OXIDASE

31. CONCLUSION
 SINGLE CELL PRODUCTION IS NOT COMMERCIALIZED METHOD.
 PROCESS CAN BE COMMERCIALIZED BY
1) RESEARCH & DEVELOPMENT IN THIS FIELD .
2) INVESTMENTS BY BIG CORPORATES.
3) PROMOTION OF THS TECHNOLOGY AT VARIOUS CONFERENCES
WORLDWIDE.
&
MOST IMPORTANT

32.  WWW.GOOGLE.COM
WWW.WIKIPEDIA.ORG
PRINCIPLE OF GENE MANIPULATION BY
PRIMROSA
BIOCHEMISTRY BY LEHNINGER.
MODERN TRENDS IN OILS & FATS
TECHNOLOGY BY OP NARULA
 RESEARCH PAPERS ON
WWW.SCIENCEDIRECT.COM