Production of biofuels from cellulosic industrial waste streams .2014-12-11  cellulosic industrial

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  • CoER : Biofuels Department of Microbiology Faculty of Natural Sciences

    UNIVERSITEIT STELLENBOSCH UNIVERSITY

    jou kennisvennoot your knowledge partner

    Production of biofuels from

    cellulosic industrial waste streams

    WH (Emile) Van Zyl & Johann F Grgens

    Department of Microbiology, University of Stellenbosch,

    Private Bag X1, Matieland 7602, South Africa

  • 2

    CoER : Biofuels (members)

    Microbiology

    Chem Eng Proc Eng

  • 3

    Lignocellulose composition

    Sugarcane bagasse Lignin 28%

    Arabinan 2%

    Xylan 25%

    Cellulose 46%

    Hexoses (fermentable) Pentoses

    (fermentable)

    Non-fermentable sugars

    high energy aromatics

    Technologies for Cellulose Conversion

  • Thermo-

    chemical

    routes

    4

    Lignocellulose

    feedstock or

    residue

    Pyrolysis Pyrolysis

    oils

    Charcoal

    Transportation

    fuels

    Catalytic

    Upgrading Refining

    Gasification

    Product gas

    or syngas

    Syngas

    Combustion for

    heat/electricity

    Product

    gas

    Synthesis Refining

    Pretreatment Enzymatic

    hydrolysis

    Fermentation

    to ethanol Distillation

    Biological route

    to cellulosic

    ethanol

    Cellulosic

    ethanol

    Residues for

    thermo-chemical

    conversion

    Consolidated BioProcessing

    Combustion

    in boiler

    High

    pressure

    steam

    Surplus heat

    Steam

    turbine Electricity Biogas

    Cooking fuel

    Lignocellulosic Biomass Conversion Options

  • 5

    Microbial conversion of cellulose to ethanol

    Sustainable Biofuels

  • 6

    Ethanol production from cellulosics

    Enzymatic hydrolysis of biomass

    Ligninases (laccases, lignin peroxidases, Mn-peroxidases)

    Cellulases (endoglucanases, cellobiohydrolases, -glucosidases)

    Hemicellulases (xylanases,-xylosidases -arabinofuranosidases -glucuronidases)

    Esterases (feruloyl esterases, coumaroyl esterases)

  • 7

    Ethanol production from cellulosics

    Spent

    material

    Pre-treatment

    Chipping

    Grinding

    Agric Res

    Woody

    Material

    Grasses

    Water

    mixing

    tank

    Steam explosion

    ~200C

    Cellulases

    Fuel

    blending

    Saccharification

    Alcohol recovery

    Distillation & dehydration

    Storage

    tank

    Yeast

    Fermentation

    Technologies for Ethanol Production

  • Steamgun Pretreatment of Lignocellulose

    8

  • 9

    Consolidated BioProcessing (CBP)

  • 10

    Pre-treatment

    Cooling &

    conditioning

    Chipping

    Grinding

    Agric Res

    Woody

    Material

    Grasses

    Water

    mixing

    tank

    Steam explosion

    ~200C

    Cellulases

    Saccharification

    Yeast

    Fermentation

    Ethanol production from cellulosics

    Spent

    material

    Fuel

    blending

    Alcohol recovery

    Distillation & dehydration

    Storage

    tank

  • Cellulolytic Yeast

    Saccharification & Fermentation

    Ethanol production from cellulosics

    11

    Spent

    material

    Fuel

    blending

    Alcohol recovery

    Distillation & dehydration

    Storage

    tank

    Pre-treatment

    Cooling &

    conditioning

    Chipping

    Grinding

    Agric Res

    Woody

    Material

    Grasses

    Water

    mixing

    tank

    Steam explosion

    ~200C

  • 12

    Xylose utilization by yeast

  • Recombinant S. cerevisiae

    G-6-P

    F-6-P

    F-1,6-dP

    DHAP

    Glycerol-3-P

    Glycerol

    G-3-P

    1,3-dP-Glyc

    3-P-Glyc

    2-P-Glyc PEP PYR

    Ethanol

    Acet

    Acetic acid

    E-4-P

    S-7-P

    Xylul-5-P

    Xylulose

    Glucose

    Ribul-5-P 6-P-Gluc 6-P-Gluclac

    Rib-5-P

    NADH + H+ + O2 + 3ADP + 3Pi

    NAD+ + H2O + 3ATP

    MITOCHONDRION

    ATP

    ADP

    NADPH NADP+ NADPH NADP+

    CO2

    ADP

    ATP

    NAD+

    NADH NAD+

    NADH

    ADP

    ATP

    NAD+

    NADH

    Pi

    XK

    RPE

    RKI

    TKL

    TKL

    TAL

    PGI

    HXK

    GPD

    ENO PYK PDC

    ALD

    NAD(P)+

    NAD(P)H

    CYTOSOL

    Glucose

    ADP

    ATP

    Xylose

    Xylitol NAD+

    NADH

    XR

    XDH

    Xylose

    NAD(P)H

    NAD(P)+

    ADP ATP CO2c

    GPDH

    PGDH

    13

  • Recombinant S. cerevisiae

    G-6-P

    F-6-P

    F-1,6-dP

    DHAP

    Glycerol-3-P

    Glycerol

    G-3-P

    1,3-dP-Glyc

    3-P-Glyc

    2-P-Glyc PEP PYR

    Ethanol

    Acet

    Acetic acid

    E-4-P

    S-7-P

    Xylul-5-P

    Xylulose

    Glucose

    Ribul-5-P 6-P-Gluc 6-P-Gluclac

    Rib-5-P

    NADH + H+ + O2 + 3ADP + 3Pi

    NAD+ + H2O + 3ATP

    MITOCHONDRION

    ATP

    ADP

    NADPH NADP+ NADPH NADP+

    CO2

    ADP

    ATP

    NAD+

    NADH NAD+

    NADH

    ADP

    ATP

    NAD+

    NADH

    Pi

    XK

    RPE

    RKI

    TKL

    TKL

    TAL

    PGI

    HXK

    GPD

    ENO PYK PDC

    ALD

    NAD(P)+

    NAD(P)H

    CYTOSOL

    Glucose

    ADP

    ATP

    Xylose

    Xylitol NAD+

    NADH

    XR

    XDH

    Xylose

    NAD(P)H

    NAD(P)+

    ADP ATP CO2c

    GPDH

    PGDH

    14

  • xylose fermentation to ethanol

    My laboratory, in collaboration with Barbel Hahn-Hgerdal in Sweden,

    were one of the first to develop a xylose-fermenting yeast (TMB3400),

    today several are available. 15

  • Recombinant S. cerevisiae

    G-6-P

    F-6-P

    F-1,6-dP

    DHAP

    Glycerol-3-P

    Glycerol

    G-3-P

    1,3-dP-Glyc

    3-P-Glyc

    2-P-Glyc PEP PYR

    Ethanol

    Acet

    Acetic acid

    E-4-P

    S-7-P

    Xylul-5-P

    Xylulose

    Glucose

    Ribul-5-P 6-P-Gluc 6-P-Gluclac

    Rib-5-P

    NADH + H+ + O2 + 3ADP + 3Pi

    NAD+ + H2O + 3ATP

    MITOCHONDRION

    ATP

    ADP

    NADPH NADP+ NADPH NADP+

    CO2

    ADP

    ATP

    NAD+

    NADH NAD+

    NADH

    ADP

    ATP

    NAD+

    NADH

    Pi

    XK

    RPE

    RKI

    TKL

    TKL

    TAL

    PGI

    HXK

    GPD

    ENO PYK PDC

    ALD

    NAD(P)+

    NAD(P)H

    CYTOSOL

    Glucose

    ADP

    ATP

    Xylose

    XI

    Xylose

    ADP ATP CO2c

    GPDH

    PGDH

    16

  • 17

    Expression of cellulases in yeast

  • 18

    Consolidated BioProcessing (CBP)

    O O

    O

    O

    O

    Glu Man Gal

    Xyl Ara

    Ethanol + CO2

    P T YFG

    Glycosyl

    Hydrolases

    Technologies for Cellulose Conversion

    Van Zyl et al. 2007. Adv. Biochem. Eng./Biotechnol. 108:205235

    La Grange et al. 2010. Appl. Microbiol. Biotechnol 87:11951208

    Lynd et al. 2002. Microbiol. Mol. Biol. Rev. 66:506577

    Lynd et al. 2005. Curr. Opin. Biotechnol. 16:577583

  • Background strain

    Appearance at 120 hrs.

    CBP strain

    0

    10

    20

    30

    40

    50

    60

    0 50 100

    Eth

    ano

    l co

    ncent

    ration

    , g/

    L

    Time (hours)

    CBP + 1 mg Xylanase

    Background +BGL + Xylanase

    Background + Cellulase + BGL + Xylanase

    Mascoma CBP technology on 18% w/w paper sludge

    Enzyme Reduction on Paper Sludge

    19

  • 20

    Converting Paper Sludge to Cellulosic Ethanol and Biogas

  • 21

    The problem

    Paper sludge is waste from paper production and recycling

    Contains degraded cellulose fibres

    15 to 50 dry tons per day per site going to landfill

    Landfill costs are high

    Environmental impact

    Limits landfill space Reduce Water to landfill (+60% moisture) Reduce methane release at landfill

  • 22

    The solution

    Transform waste into green transport fuel Mitigate environmental impacts with

    attractive returns

    Convert paper sludge to bio-ethanol with CBP Avoid high enzyme costs (critical for

    viability) Reduce waste to landfill (50%) (GHG, space) Reduce water content and water to landfill Does not impact on food production Ethanol is a green replacement for fossil fuel

  • 23

    The Paper sludge Pilot

  • 24

    The Financial Summary

    Dependent on

    Equipment cost Paper sludge cost Ash content of

    sludge Production scale

    Selling price of

    ethanol

    Assumptions

    ZAR 65m (ZAR880/ton) 10%

    50 dry tons per day 5.4 ML/annum EtOH ZAR5 7 per litre

    Returns

    NPV = ZAR 26.8m Project IRR = 27% Payback = 3 years Viable with 50%

    reductions

    For more information, contact : Johann Grgens -

    jgorgens@sun.ac.za Mobile: +27 82 448 4648

  • 25

    Feedstock:

    Ngodwana Raw

    Virgin Paper Mill Sludge

    Springs Raw

    Recycled Paper Mill Sludge

    Springs Processed

    Residues from ethanol fermentation of Raw Springs

    Inoculum:

    Mesophilic mixed c