Microalgae for valorization of industrial wastes from pulp/paper mills and oil refineries

1. Microalgae for valorization of industrial wastes from pulp/paper mills and oil refineries Algae Biomass Summit 2014 A. Kuehnle, N.A. Nolasco, M. Perez, M. Johnson

2. Overview: Wood Waste from Pulp/Paper Mills Under USDA BRDI grant, Domtar teamed with KAS to valorize underutilized wood waste using microbial conversion into bioproducts. • Domtar • Largest uncoated freesheet (UFS) and paper grade pulp manufacturer in North America • Recently added diversity to its product mix, ventured into consumer products • Goal: “Fuel to feed” • Hog fuel into feed (and higher value algal bioproducts) • Wood hydrolysates produced by CSI, Cellulose Sciences International Bleached Southern Pine hydrolysate Southern Hardwood Chips hydrolysate

3. Process • Screen and select strains from KAS collection for growth on mixed sugars, tolerance to inhibitors in wood hydrolysates • Establish algal biomass productivity and sugar utilization (HPLC) using aerobic fermentation • Nutrient medium targeted biomass productivity (feed value and co-products) • Batch fermentation conditions • T= 30°C, pH= 7.0, agitation = 300 rpm, DO2 = 100%, air = 3.0 L/min • Hawaiian species, including Chlorella and Scenedesmus

4. Wood hydrolysates from CSI • Hydrolysates profile: Salinity ranged 3.3% – 4.2%, pH: 5.5; undefined components • Screening and controls: Standardized with C5 and C6 sugar equivalents

5. Growth Varied with Each Type of Wood hydrolysate • All five wood hydrolysates supported growth of algae

6. Wood Hydrolysates Outperform Model Sugars • A 1.6-fold higher biomass productivity (2.87 g/L-day), was obtained with bleached southern pine (BSP) hydrolysate than with C5 and C6 model sugars alone (1.7 g/L-day).

7. Other Outcomes • C5 sugar effect • • Some strains thrive in hydrolysate vs in equivalent glucose alone • Excellent sugar utilization, varies with hydrolysate and impurities • 2:1 sugars : biomass ratio (SHC), EtOH present as impurity • Beneficial non-sugar components present in CSI hydrolysates Specific sugar utilization of algae in two wood hydrolysates 5:1 2:1 SHC BSP

8. Concluding Thoughts • Opportunity to develop mill-based strategies matching hog fuel with strains • Can select preferred strain with lowest sugar requirement • Select strain that benefits from non-sugar components present • Select strains to utilize both the C5 and C6 sugars • Co-cultivate strains to match wood composition, efficiency • Product options for biomass • Align volumes with off-take potential • Preferably match mill products with regional processors and off-takers • Next steps • Validate metrics at next scale to meet targets in financial models

9. Overview: Wastewaterfrom Oil Refineries • Algae-wastewater pilot facility was designed, constructed and operated in 2013-2014 at Chevron Hawaii to demonstrate nitrogen and phosphorous reduction in refinery wastewater • Reduction in total suspended solids (TSS) in the effluent was also addressed, using tangential flow filtration • Utilized waste CO2 from refinery’s hydrogen formation plant, as previously demonstrated by KAS in EPA-award winning project

10. Algae Pilot Plant PBR and Simulation Tank • PBR used Clear-Flex polyurethane tubing with sterile fittings fabricated to spec for KAS by Siftex (CT) • Simulation tank (10-feet deep) used basal pneumatic mixing with plant air and CO2 on discontinuous basis • System designed for future deployment into existing basins for minimal footprint

11. Effluent and TestProtocol Chlorella-like Bacteria Algae Pilot Scenedesmussp. Initial TestAmerica: NH3-N, NO3-NO2 N, TKN, Total N, Total P KAS: NH3-N, NO3-N, PO4-P, algal count, bacterial CFU, DW (or TSS), DO, pH, temp Final TestAmerica: NH3-N, NO3-NO2 N, TKN, Total N, Total P KAS: NH3-N, NO3-N, PO4-P, algal count, bacterial CFU, DW (or TSS), DO, pH, temp During the Test (KAS) NH3-N, NO3-N, PO4-P algal count, bacterial CFU, DW (or TSS), DO, pH, temp • •Triplicate grab samples- before and after treatment- were analyzed by TestAmerica

12. Wastewater rich in ammonia N • N and P levels significantly reduced in 3 d • 97% reduction in Ammonia N, 69% reduction in Total N, 90% reduction in Total P Reduction: 69% 97% 90% Total N NH3 N Total P

13. Scenedesmus thrived in the wastewater, compatible with the other microbes • Previous work by KAS with this and other tested organisms showed utilization of mixed N sources, and variable amounts of NH3 and NO3 • Very important for fluctuating oil refinery wastewater profiles • Met end objective of using mixed microbes to best polish the wastewater KAS740

14. Other Outcomes • Hollow fiber filtration removed 99+% of total suspended solids; suited to solids recycle as needed • Basal pneumatic mixing system effective for deep ponds, promising for use in existing basins • For managing nutrient loads and suspended solids: • Results support final polishing of wastewater using KAS’s basis of design and P&ID solution • Used off-the-shelf industrial equipment and components in innovative manner for ease of deployment

15. Concluding Thoughts • Using KAS solution, possibility to address targets in both downstream and upstream pits/basins of the O&G industry • Potential $10 B O&G wastewater market in U.S. by 2025 • Additional validation, deployment with engineering firm • Difficult to sell into this market without a partner