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Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium

Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium. Javier M. Naranjo , John A. Posada, Juan C. Higuita , Carlos A. Cardona Instituto de Biotecnología y Agroindustria, Universidad Nacional de Colombia sede Manizales, .

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Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium

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  1. Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium Javier M. Naranjo, John A. Posada, Juan C. Higuita, Carlos A. Cardona Instituto de Biotecnología y Agroindustria, Universidad Nacional de Colombia sede Manizales, Betreuer: Benjamen Stenger Ammar Abdulmughni

  2. Overview • Introduction Polyhydroxyakanoate (PHA): Structure, Biosynthesis Production • Aimofthissubject • Results • Summary

  3. Polyhydroxyakanoate (PHA) • PHAs are linear polyesters • Manyprokaryotescansynthesizeandstore PHAs in form ofgranules (upto 80 % of total bacterialweight) • PHAs havesimilarmechanicalpropertiestoconventionalplastics • Theyaregenerallybiodegradable

  4. Polyhydroxyakanoate (PHA) • The biosynthesis of PHA : certain deficiency conditions and excess supply of carbon sources • The mostcommon/produced form of PHA ispolyhydroxybutyrate, PHB • Likepolypropyleneorpolyethylene. PHB

  5. Biosynthesis of PHB

  6. PHB Production • Productioncostsareheigh:  PHB accumulateintercellularly: - Useof a solvent - thermal treatment of biomass, enzymatic digestion and washing with an surfactant a self-disruptivestrain.

  7. The lysissystemofBacillusamyloliquefaciens • phage was insertedintoshuttlevector • The expression of a target gene is inhibited by • glucose •  When the glucose concentration approached • zero, self-disruption is induced

  8. PHB Production • Productioncostsareheigh: carbonsources The carbon source could account for 25–45% of the total production costs Cheapercarbonsources

  9. PHB Production • Agroindustrial wastesareattractivecandidatesassubstrates:  lowpricesandheighavailability  solving an environmental problem • A variety of microorganisms are able to produce PHB from diverse agroindustrial wastes: Methylobacteriumrhodesianum(Borman et al., 1999) Cupriavidusnecator(Cavalheiro et al., 2009)  E. coli CT1061 (Nikelet al., 2008)

  10. Glyceroliscarbonsource • Crudeglycerolis a co-product in theproductionofbiodiesel • Glycerolis an importantindustrialfeedstock : - food, drugs, cosmetics, pharmaceuticals, textile andtobaccoindustries • Glycerolcanbeusedascarbonsource in microbiologicalprocesses • Polyhydroxyakanoate (PHA) productionis an interestingbiologicaltransformationofglycerol.

  11. PHB Production • Currently, PHB is produced at an industrial scale using Gram negative bacteria  lipopolysaccharides (LPS) which co-purify with the PHAs and induce a strong immunogenic reaction (Valappil et al., 2007) Bacillusmegaterium

  12. Aim • Techno-economicanalysisof PHB production: - Glyceroltransformationinto PHB (usingBacillusmegaterium) - Glycerolor Glucose assubstrate - Productivityat different conditions

  13. Results • Different initialconcentrationofglycerol PHB Production Biomass The fermentation conditions: temperature 30 °C, air flow 12 l/min and uncontrolled pH

  14. Results • Different temperaturesoffermentation PHB Production Biomass The fermentation conditions: initial glycerol concentration of 20 g/l, air flow 12 l/min, and uncontrolled pH

  15. Results • PHB productionusingglucoseorglycerol VerySimilaryields

  16. Results • Batch cultivation: The fermentations to produce PHB were carried out for 42 h in a 3.7 l Lab Fermen

  17. Results • Economicanalysis: • The current sale prices are between 3.1 and 4.4 USD/kg

  18. Summary • PHAs are linear polyestersandcanbesynthesizedanditnracellularlystoredbymanyprokaryotes. • The mostproduced form of PHA is PHB • PHAs havesimilarmechanicalpropertiestoconventionalplastics • Productioncostsareheigh • PHB ProductionusingB. megaterium, andglycerolascarbonsource • The results confirm the ability of B. megaterium to use glycerol as the only carbon source.

  19. References • Valorization of glycerol through the production of biopolymers: The PHB caseusingBacillusmegaterium, Javier M. Naranjo, John A. Posada, Juan C. Higuita, Carlos A. Cardona, Bioresource Technology 133 (2013) 38–44. • Polyhydroxybutyratesynthesis on biodieselwastewaterusingmixedmicrobialconsortia, Zachary T. Dobroth a, Shengjun Hub, Erik R. Coats a, Armando G. McDonaldBioresource Technology 102 (2011) 3352–3359 • Large-scale production and efficient recovery of PHB with desirable material properties, from the newly characterised Bacillus cereus SPV, Valappil SP, Misra SK, Boccaccini AR, Keshavarz T, Bucke C, Roy I., J Biotechnol. 2007 Nov 1;132(3):251-8 • Construction of self-disruptive Bacillus megaterium in response to substrate exhaustion for polyhydroxybutyrate production. Hori K, Kaneko M, Tanji Y, Xing XH, Unno H. Graduate School of Bioscience and Biotechnology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.

  20. THANK YOU

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