biocrude production by activated sludge microorganisms
TRANSCRIPT
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Biocrude Production by Activated Sludgeusing amended Pulp and Paper wastewater
as substrate
Kamal Lamichhane , Andro Mondala, Rafael Hernandez, Todd French, Magan Green,Linda McFarland, William Holmes
Dave C. Swalm School of Chemical Engineering, Mississippi State University
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Content
Objective
Introduction
Materials and methods
Results and
discussions
Conclusion
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Objective
The objectives of this study were to:
1. Test pulp and paper waste water to grow
microorganisms capable of lipid
accumulation, and measure the lipidaccumulation and biodiesel yield
2. Compare the lipid and biodiesel yield
using pulp and paper wastewater with
synthetic wastewater
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Wastewater Treatment Plant for Enhancement of lipid
yield
Screening/grit
removal
Aeration tank
Clarifie
r
Air
Air
Clarifie
r
Domestic/industrialWastewater
influents Effluent
Oily
sludge tolipid
extraction
steps
Belt press
Activated
sludge
Extruded
water
Clarifier
Lipid
accumulation
chamber
Carbon and
nutrient
sources
Recycle
Holdingtanks
Primarysludge to
anaerobic
digestion or
lipid
extraction
Effluent
Waste
sludge
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Pulp and Paper wastewater
Approximately 1551 billiongallons of wastewaterproduced annually in theUnited States1
For every ton of pulp
produced, 17,000 galloonsof industrial process waterrequired2
Almost half of the total
waste is produced in thesouthern regionFig1: International Paper, Vicksburg, MS
1Effluent Limitations and Guidelines, Pretreatment Standards, and New Source Performance Standards: Pulp, Paper and
Paperboard Category; National Emission Standards for Hazardous Air Pollutants for Source Category; Pulp and Paper
Production; Proposed Rule, Dec. 17, 1993
2. http://chlorinefreeproducts.org/images/Article3.pdf
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Pulp and Paper Wastewater:
characteristics and common
contaminants Characteristics
High COD (500-1500 mg/L)
Dark brown coloration
Adsorbable organic halide
Toxic pollutants
Common Contaminants
Sodium carbonate
Sodium hydroxide
Sulfates Chlorine dioxide
Calcium oxide
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Manufacturing Process
Wood
PreparationCooking(NaOH
and Na2S)
Pulp
Washing
Pulp
Screening
Bleaching
Wood Preparation: removes bark
Cooking: most of the lignin removed from the
wood
Pulp washing: wash out the cooking chemicals
and lignin from wood fiber
Pulp screening: remove the uncooked fibers
Bleaching: two stage bleaching-a) Pulp treated with NaOH in presence of
O2to break down the polymer
b) Pulp is treated with ClO2to remove the
remaining the lignin
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Wastewater From Pulp and Paper
Mills
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Wastewater Treatment
Process
Fig 2: Flow Scheme of general Pulp and Paper Wastewater Treatment Process*
*Ince, Bahar K., Cetecioglu, Zeynep., Ince, Orhan (2011).Pollution prevention in the pulp andpaper Industry. pp 231.
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Activated Sludge Process
Biological method that usesmicroorganisms to feed on
organic contaminants in
wastewater
The microorganisms form flocand settle at the bottom
leaving clear liquid at the top
The top liquid is free of
organic materials andsuspended solids*
*Expanding the Activated Sludge Process (2003). Retrieved September 23, 2012 from
http://www.nesc.wvu.edu/pdf/ww/publications/pipline/pl_sp03.pdf
Fig3: typical activated sludge processhttp://www.enotes.com/topic/Activated_sludge
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Oleaginous microorganisms
Activated Sludge contains amixed microbial community
Oleaginous microorganisms in
activated sludge can
accumulate a high percentageof lipids
Microorganisms require water
with high organic content (high
C:N ratio) to grow
Pulp and paper wastewatercould be a suitable growth
media for microorganisms
specialized in biocrude
production
Fig 4: Cells of an Oleaginous yeast*
*. Wynn, James P. Ratledge, Colin. (2005) Oils from Microorganisms pp 123.
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Wastewater Initial Characteristics
Fig 5: Different types of pulp andpaper wastewater
Characteristi
csPulp Wash
Mixed
WastewaterPond Effluent
Process
derived from
Wood pulping,
bleaching, and
washing.
Combined
wastewater
streams
from mill.
Effluent from
anaerobic
settling pond.
pH 2.25 6.26 8.59
COD (mg/L) 1478 1361 410
Ammonium
(mg/L)3.95 5.90 2.83
Sulfate
(mg/L)1400 840 400
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Materials and Methods
Following fermentation experiment wasset up in the bioflo 310 fermenter
2.4 L of the wastewater
20% Activated sludge ( 600 ml)
60 g/L sugar 1.3 g/L Ammonium chloride
1.5 g/L of Potassium phosphate and
1g/L of sodium phosphate
The initial pH was set to 6.5 and agitation
was maintained at 300 rpm and changedto maintain the DO at 60% saturation
Fig 5: Bioflo 310
Fermenter
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Experimental ProcedureActivated
Sludge+
wastewater
Fermentation
Centrifuge
NH4+, COD, sugar Freeze dry
Bligh and
Dyer+TurboVap
Transesterification
GCFID
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Kinetics: Biomass growth
Mathematical modeling was done following the methods by Satturet al. (1991), Karanth et al. (1991) and Mondala et al. (2011)
The biomass growth was determined using the logistic model given
by Weiss and Ollis
max
max
max0
maxmax
max
max
max
max
slopewithlinestraightagivestvslnofplot
ln1lnt
g,Reaggangin
biomassMaximum
rategrowthspecificMaximum
biomasslipidnonofionConcentratX(t)where,
)(1)(
XX
X
XX
X
X
X
X
X
tX
tXdt
dX
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Kinetics: Lipid Accumulation
Lipid accumulation was modeled using Luedking-Piretequation
Vs. ( X X 0) gives straight line
with slope mPlotting
After linearization, the final values of the kinetic parameters were
estimated by non-linear regression using the Levenberg-Marquardt
method in Polymath 6.1
Po = Initial Lipid concentrationn= stationary phase constant
m= growth phase constant
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Kinetics: Substrate
consumption Substrate consumption was modeled by a Luedeking- Piret
equation
XKdt
dP
Y
m
dt
dX
Ydt
dSe
SPSX
//
1e
SP
KY
n
/
SPSX Y
m
Y//
1
O
O
O XtX
tX
Xt
XStS
)()(
ln)( maxmax
max
S0, X0= initial concentration of sugar, and non lipid biomass respectively
max= Maximum specific growth rate
Ke= Maintenance coefficient,= constants used to predict residual sugar maintenance coefficient
Yx= biomass yield based on sugar consumed
Yp= Lipid yield based on sugar consumed
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Results : Pulp Wash
0
50
100
150
200
250
300
350
400
0
10
20
30
40
50
60
0 24 48 72 96 120 144 168
Ammonium(m
g/L)
Biomass,
Lipid,
Glucose
Time(hr)
biomass (g/L)
glucose(g/L)
lipid (%w/w)
ammonium(mg/L)
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Results : Pond Water
0
50
100
150
200
250
300
350
400
450
0
10
20
30
40
50
60
0 24 48 72 96 120 144 168
Ammonium
Biomass,
Lipid,G
lucose
Time (hr)
biomass(g/L)
lipid(%w/w)
glucose(g/L)
ammonium (mg/L)
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Results: Mixed wastewater
0
50
100
150
200
250
300
350
400
0
10
20
30
40
50
60
0 24 48 72 96 120 144 168
Ammonium
Biomass,
Lipid,
Glu
cose
Time (hr)
biomass (g/L)
glucose (g/L)
lipid (%w/w)
ammonium (mg/L)
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Result: Lipid and Fames
Wastewater
Biocrude
(%w/w)
Biodiesel
(%w/w)
Pulp Wash 16.04 6.22
Pond Water 29.59 7.45
Mixed wastewater 10.77 5.9
Fatty Acid Pulp Wash Pond Water Mixed Waste
Myristic (C14:0) % 1.16 0.38 1.03
Palmitic (C16:0) % 19.56 19.4 19.35
Palmitoleic (C16:1) % 3.42 3.44 3.9
Stearic (C18:0) % 11.4 11.03 11.37
Oleic (C18:1) % 44.94 50.26 44.82
Linoleic (C18:2) % 14.93 13.15 15.19Linolenic (C18:3) % 2.41 1.83 1.93
Arachidic (C20:0) % 0.69 0.36 0.52
Lignoceric (C24:0) % 0 0.15 1.64
Table: Fatty Acid Profile
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Kinetic Model Parameters
Biomass growth
Xo 7.7347
Xmax 15.110
max
0.008837
R2 0.7676
Lipid Accumulation
m 0.1352
n 0.0004697
R2 0.8054
Sugar Consumption
1.5849
0.021948
R2 0.9622
Lipid Accumulation
m 0.958
n 0.00304
R2 0.80158
Biomass growth
Xo 8.67
Xmax 17.05
max
0.003528
R2 0.40043
Sugar Consumption
8.5732
0.0172246
R2 0.90832
Lipid
Accumulation
m 0.0319
n 0.0005047
R2 0.76337
Biomass growth
Xo 8.29879
Xmax 19.2200
max
0.012329
R2 0.82707
Sugar Consumption
1.8159
0.0185692
R2 0.9598
Pulp Wash Pond Water Mixed Waste
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Remarks
1. All of the three wastewaters were suitable foroleaginous microbial growth when amended with
sugar and other nutrients
2. Microorganisms growing in pond water accumulated
most lipid (29.59 % CDW) at day 7 compared to pulpwash (11.69%) and mixed wastewater (8.19%)
3. The biodiesel yield was around 6-7 % W/W for all the
wastewater
4. Fatty acid profile was similar to those obtained fromdomestic wastewater
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Future Work
Optimization of fermentation conditions andinvestigation of fed-batch or continuous modes.
Identification of possible toxic compounds and
determination of their fate in activated sludge
lipid accumulation cultures.
Correlate BOD and COD reduction with
biocrude production.
Utilization of pulp and paper mill solid wastesand fiber as source of fermentable sugars.
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