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

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OVERVIEW: WOOD WASTE FROM PULP/PAPER MILLS

Bleached Southern Pine hydrolysate

Southern Hardwood Chips hydrolysate

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

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

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Hydrolysates profile: Salinity ranged 3.3% – 4.2%, pH: 5.5; undefined components Screening and controls: Standardized with C5 and C6 sugar equivalents

WOOD HYDROLYSATES FROM CSI

GROWTH VARIED WITH EACH TYPE OF WOOD HYDROLYSATE

All five wood hydrolysates supported growth of algae

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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).

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OTHER OUTCOMES

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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

SHCBSP

Specific sugar utilization of algae in two wood hydrolysates

2:1

5:1

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

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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

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OVERVIEW: WASTEWATER FROM OIL REFINERIES

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

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EFFLUENT AND TEST PROTOCOL

Algae Pilot

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

During the Test (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

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Chlorella-like

Scenedesmus sp.

Bacteria

•Triplicate grab samples- before and after treatment- were analyzed by TestAmerica

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

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Initial Final

Total N

NH3N

Total P

69%

Reduction:

97% 90%

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

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KAS740

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

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OTHER OUTCOMES

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 2025Additional validation, deployment with engineering firm

Difficult to sell into this market without a partner

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CONCLUDING THOUGHTS