Project 3.4: PRODUCTION OF POLYHYDROXYALKANOATES (BIO-PLASTICS) FROM SUGAR CANE TRASH AND VINASSE

1. Project 3.4: PRODUCTION OF POLYHYDROXYALKANOATES (BIO-PLASTICS)FROM SUGAR CANE TRASH AND VINASSE Mauritius Sugarcane Industry Research Institute Mid-term Review - 2 October 2012

2. Background • What? Bioplastics are plastics in which all carbon is derived from renewable feedstocks. Polyhydroxyalkanoates (PHAs) are a group of bioplastics that are completely synthesized by microorganisms

3. Background • Why? • Produced from renewable resources v/s petroleum-based • Reduced fossil-fuel use & CO2 emission: reduced environmental impact. • Biodegradability • Concerns about human health • Potential for sustainable industrial production • Wide application (packaging material, agriculture, biomedical etc.)

4. Background • Why not? High production cost in comparison to petroleum-based plastics : the high raw material cost high recovery cost low PHA yield

5. 2005: Project proposal first formulated emphasis was on use of sugarcane bagasse as feedstock Mar 2011: Amended project proposal was presented to the SAG sugarcane trash and vinasse were included as likely feedstock SAGrecommendations: To revisit Project Desktop study Aug 2011: Desktop study completed Revised Project proposal submitted

6. Desktop study: • Current use and composition profile of vinasse. • Vinasse as a potential feedstock for production of PHA. • Sugarcane trash as potential feedstock for the production of PHA. • Pre-treatment and hydrolysis of the lignocellulosic biomass from sugarcane. • Theoretical PHA yield from sugarcane biomass and vinasse. • Recommendations

7. Key findings from desktop study: • low PHA production potential of vinasse - its use in a stand-alone PHA production plant unlikely to be feasible. • As much PHA may potentially be obtained from sugarcane trash on a per hectare basis as from bagasse. • Vinasse may be considered as part of a process integrated into a trash-based PHA production plant.

8. PROJECT GOAL:To diversify & expand the revenue base of the sugar sector in Mauritius and other ACP countries Purpose:To develop a cost-effective technology for the production of PHA from unused/cheap carbon sources from sugarcane.

9. Researchsteps • Efficient conversion of the cellulose / hemicelluloses in the sugarcane trash/vinasse into simple sugars Isolation, identification and selection of PHA-producing bacteria • Optimization of media and conditions for maximum bacterial growth and then maximum PHA production • Optimization of extraction procedures and efficiency of PHA recovery

10. Proposed scheme for PHA production using vinasse and sugarcane trash as feedstock Lignocellulosic Biomass (trash) Optimization is required Vinasse Sugars, VFA’s Prehydrolysate Separation Pre-treatment • Production of PHA-rich microbial biomass • Separation Effluent & residual biomass Extraction & Purification of PHA • Hydrolysis SOLIDS Hydrolysate Separation PHA polymer SOLIDS Fertilizer • Fuel

11. ACTIVITIES PLANNED FOR 2011-2012 • 1. Purchase of equipment: • January 2012 : International Open Tender published in for the purchase of • 1 liquid chromatograph-mass spectrometer 1 gas chromatograph-mass spectrometer, and • 1 gel permeation chromatograph. • June 2012 : Evaluation of tenders received completed • Award of contract awaiting release of funds • Order of 12 items of smaller equipment for Microbiology laboratory by Negotiated Procedure awaiting release of funds

12. ACTIVITIES PLANNED FOR 2011-2012 Recruitment and training of scientists February 2012 : Two graduates recruited on contractual basis. In-house training on microbiology and chemistry aspects. Delay in acquisition of new equipment has delayed training 23 November to 3 December 2011: Two scientists from MSIRI visited the Department of Bioprocess Technology, University Putra Malaysia (UPM) and the Ecobiomaterials Research Laboratory, University of Science Malaysia (USM)

13. ACTIVITIES PLANNED FOR 2011-2012 Isolation, identification and selection of PHA-producing bacteria: Bacterial isolates (46) collected and purified from different niches (vinasse, sugarcane trash, soil, refinery mud) as well as reference isolates known to produce bioplastics (e.g. Ralstoniaeutropha, Azospirillumbrasilense, Rhizobium sp, Bacillus sp) are being evaluated for their ability to produce bioplastics Optimization of bacterial growth and of PHA biosynthesis : Different nutrient media are being evaluated in shake-flask culture and under different conditions of shaking and incubation temperature so as to optimize the bulking of bacterial cells.

14. ACTIVITIES PLANNED FOR 2011-2012 Optimization of PHA recovery and purification: The evaluation of different PHA extraction protocols for recovery of PHA from bacterial cells, including the use solvents, non-solvents and mechanical means, has been initiated. Characterization of sugarcane trash, vinassse: Sugarcane trash and vinasse have only partially been characterized for a number of simple parameters but the quantification of convertible sugars, volatile fatty acids and other microbially-assimilable substances is awaiting the acquisition of new equipment (HPLC system)

15. ACTIVITIES PLANNED FOR 2011-2012 Pre-treatment and hydrolysis of biomass , Characterization of hyrolysates: This activity will be performed once the necessary equipment are received.

16. Plan of activities

17. Plan of activities

18. Plan of activities

19. Expected outcome To set the basis for establishment of a future bioplastic production facility based on unused/cheap sources of carbon from sugarcane. Capacity building; acquisition of state-of the-art equipment. Beneficiaries All sugar producing countries looking for a diversification of the sugar industry.

20. Acknowledgements

21. Thank you for your attention

26. Theoretical PHA yield from biomass and vinasse.