Developing Cost Effective Biological Removal Technology for Selenium and Nitrate from Flue Gas Desulfurization (FGD) Wastewater from an Existing Power Generating

Facility

Dr. Sanju A. SanjayaEnergy and Environmental Science Institute

West Virginia State University (WVSU) sanjaya@wvstateu.edu

DOE NETL Project Review Meeting, April 09 – 11, 2019

Outline

•Project Description and Objectives •Background Information•Project Update•Preparing Project for Next Steps•Concluding Remarks

ObjectivesThe overall goal of our project is to investigate and determine a technically feasible and cost-effective process for designing photosynthetic organisms capable of sequestering Se and nitrates from FGD wastewater. To realize this goal, we have chosen to focus on the following 2 objectives:

(1) Investigate changes in transcripts and metabolism in algae and plants in response to FGD wastewater.

(2) Explore biotechnological strategies to increase sequestration of Se and nitrates in biomass to improve agricultural productivity.

Academic objective: to enhance student hands-on experience and participation in STEM research and education

Thermoelectric Power and Freshwater Use

4

Freshwater consumption is projected to increase further with the implementation of carbon capture technologies

Background Information

• Flue Gas Desulfurization (FGD) treatment is incorporated in most coal burning power generation plants to remove sulfur dioxide and various oxides of nitrogen by either wet/dry scrubbing.

• Large-scale coal fired thermoelectric plants in the USA consumes significant volumes of freshwater and generate considerable amounts of FGD wastewater.

• Wet scrubber blowdown often contains heavy metals (selenium, chromium, mercury etc.,), and nitrates in harmful concentrations.

• These constitute a major challenge for utilities and a major concern for environmental regulators.

Green algae and plants have the natural ability to degrade inorganic Se and nitrates

Pilon-Smits and Quinn in R. Hell and R.-R. Mendel (eds.), Cell Biology of Metals and Nutrients, Plant Cell Monographs 17,DOI 10.1007/978-3-642-10613-2_10, # Springer-Verlag Berlin Heidelberg 2010

Overview of the movement and metabolic conversion of Se by plants and their ecological implications

In collaboration with John Amos power plant through Liberty Hydro and obtained treated and untreated FGD samples

Project Update

C

FGD

T-FGD

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101520253035404550

Week-1 week-2 Week-3

Biom

ass (

mg

Fres

h W

T /2

50 m

l cu

lture

)

C FGD T-FGD

Week-1 Week-2 Week-3

Growth of Algae in culture medium supplemented with FGD and T-FGD wastewater for week-1 to 3; B. Algae biomass measurement for week-1 to 3, data were derived from threereplications. Data indicate that algal biomass growth rate was higher in mediumsupplemented with FGD and T-FGD wastewater. C: Control; FGD: Flue gas desulfurizationwastewater; T-FGD: Physical/Chemical treated flue gas desulfurization wastewater.

A B

Effect of FGD wastewater on freshwater algae biomass

T-FGD T-FGD T-FGD T-FGD

FGD FGD FGD FGD

C C C C

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Biom

ass (

mg

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h W

T/3

seed

lings

)

C

FGD

T-FGD

A. Growth of Arabidopsis in soil watered with FGD and T-FGD wastewater for week-1 to 3; B. Plant biomass measurement for week-1 to 3, data were derived from three replications. Data indicate that plant biomass growth rate was higher in plants watered with T-FGD wastewater. C: Control; FGD: Flue gas desulfurization wastewater; T-FGD: Physical/Chemical treated flue gas desulfurization wastewater.

Effect of FGD wastewater on plant biomass (Arabidopsis thaliana)

Effect of FGD wastewater on Duckweed biomass

Control FGD T-FGD

Treatment of Duckweed with FGD wastewater increases weight

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Control 24h FGD 48h FGD 72h FGD

Biom

as D

ry W

T (m

g)

Effect of FGD Wastewater on Lipid Production in Duckweed

TAG

DAG

Triacylglycerols

FGD wastewater treatment promote the accumulation of lipid droplets in Duckweed

72hr FGD72hr control

Quantification of Total TAG content in Duckweed using GC/MS

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0.5

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1.5

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Control FGD 24hr FGD 48hr FGD 72hr

TAG

(mg/

Dry

Wei

ght)

Analysis of Different Lipid Species Using GC/MS

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G (m

g/DW

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Control FGD 24hr FGD 48hr FGD 72hr

TAG/Oils Are the Most Energy-Dense Plant Products

Glucose (CHOH)n

Oleic acidCH3(CH2)nCOOH

More ReducedCarbon

Ohlrogge and Chapman, 2011, The Biochemical Society, April, 34-38

Oils have twice the energy content of carbohydrate and proteins

Chart1

Triacylglycerol

Protein

Carbohydrate

kCal/gm

Energy Content (kCal/gm)

9

4

4

Sheet1

TriacylglycerolProteinCarbohydrate

944

Analysis of Lipid Composition in Duckweed

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DGDG MGDG PG LysoPG LysoPC LysoPE PC PE PI PS PA

mol

%

Control FGD 24hr FGD 48hr FGD 72hr

Change in the Total Protein Content

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0.2

0.4

0.6

0.8

1

1.2

1.4

Control 24h WFGD 48h WFGD 72h WFGD

Tota

l Sol

uble

Pro

tein

s (m

g/m

l)

Wang et al., 2009. Nat Rev Genet; 10(1): 57–63

Perform post-sequence analysis and qRT-PCR analysis

RNA-Seq Workflow

Heat map of top differentially expressed genes in DuckweedContrast between C24 VS FGD24 Contrast between C48 VS FGD48 Contrast between C72 VS FGD72

Differentially expressed Lipid related genes heat maps in Duckweed

Contrast between C24 VS FGD24 Contrast between C48 VS FGD48 Contrast between C72 VS FGD72

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0 24 48 72

#1

Rela

tive

fold

cha

nge

Hrs after treatment

050

100150200250300350

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#2

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Hrs after treatment

00.20.40.60.8

11.2

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#3

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Hrs after treatment

00.20.40.60.8

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#9

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Hrs after treatment

00.20.40.60.8

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#7

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Hrs after treatment

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#5

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Hrs after treatment

RT-qPCR analysis of Fatty acid biosynthesis genes in Duckweed

TAG Lipase Aminophospholipid ATPase

Oleate Desaturase

Long-Chain Acyl-CoA Synthetase

Oxo-Phytodienoic Acid Reductase Golden2-like

Perform functional analysis of candidate genes in model system

• Short generation time-6-8 weeks from seed-seed• Small size• Wider adaptability• Self-fertilization• Susceptibility to Agrobacterium infection• Small genome size -125 million base pairs• Large collection of T-DNA

Effect of FDG wastewater on coal mine soil and establishment of energy crop

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0.12

Wei

ght (

mg)

Ag Soil Coal mine Soil +FGD

Coal mine soil supplemented with FGD wastewater increase seed yield

Schematic representation of binary vectors used for

transgenic plants establishment

1 T35S AtNRT135S 35SHptIILB RB

nosT35S 35SDsRed

2 T35S AtSULTR135S 35SHptII nosT35S 35SDsRed

3 nos 35SAtSULTR1T35S AtNRT135S 35SHptII nosT35S 35SDsRed

Current status:

Homozygous lines are established…

Learning Water Quality in a Class Room

Oral and Poster presentation in 1890 ARD symposium, Jacksonville, FL 2019Oral presentation in ASPB Midwest section, WVU symposium, 2019Oral presentation in Gordon Research Conference on Plant lipids, Galveston, TX, 2019

Studying the Effect of Soil on Plant Physiology

30

Preparing Project for Next Steps

• FGD wastewater treatment triggered the accumulation of useful fatty acids in the Duckweed, the potential of this technology need to be investigated.

• Supplementation of FGD wastewater in coal mine soil promoted the growth of energy crops, use of FGD wastewater in re-use in Agriculture needs to be determined.

Concluding Remarks

• Completed the RNA Seq experiments for duckweeds treated with FGD wastewater for different time course and identified candidate genes involved in fatty acids and metal sequestration.

• Demonstrated the re-use of FGD wastewater in Agriculture on coal mine soils using bioenergy crop.

Acknowledgement Collaboration

Josh Davidson; Bagya Muthan; Joseph Baird; Tawna Heath; Thanh Nauyen and Krishna Challa, Bhikash Deo, Subhiksha Khathiwada, and Justin Bradley

Funding:

Liberty Hydro, Inc.,

Bio-Chem Testing, Inc.,

Developing Cost Effective Biological Removal Technology for Selenium and Nitrate from Flue Gas Desulfurization (FGD) Wastewater from an Existing Power Generating FacilityOutlineObjectivesThermoelectric Power and Freshwater Use�Background InformationSlide Number 6Slide Number 7Slide Number 8Slide Number 9Slide Number 10Slide Number 11Treatment of Duckweed with FGD wastewater increases weightEffect of FGD Wastewater on Lipid Production in DuckweedFGD wastewater treatment promote the accumulation of lipid droplets in DuckweedQuantification of Total TAG content in Duckweed using GC/MSAnalysis of Different Lipid Species Using GC/MSTAG/Oils Are the Most Energy-Dense Plant ProductsAnalysis of Lipid Composition in DuckweedChange in the Total Protein ContentSlide Number 20Slide Number 21Slide Number 22Slide Number 23Slide Number 24Slide Number 25Slide Number 26Slide Number 27Slide Number 28Learning Water Quality in a Class RoomStudying the Effect of Soil on Plant PhysiologyPreparing Project for Next Steps�Concluding Remarks�Acknowledgement