NREL is a national laboratory of the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.

High Throughput Pretreatment and Enzyme Hydrolysis of Biomass: Screening Recalcitrance in Large Sample Populations

Presented by

Stephen R. Decker

1st International Conference on Lignocellulosic Ethanol

Copenhagen, Denmark

October 13-15, 2010

NREL/PR-2700-49599

BioEnergy Science Center

Oak Ridge National Laboratory

University of GeorgiaUniversity of Tennessee

Dartmouth CollegeGeorgia Institute of Technology

West Virginia UniversityArborGen, LLC

Ceres, Incorporated

Mascoma CorporationVerenium Corporation

Samuel Roberts Noble FoundationNational RenewableEnergy LaboratoryBrookhaven NationalLaboratoryUniversity of California–Riverside Cornell University Washington State UniversityUniversity of Minnesota North Carolina State UniversityVirginia Polytechnic InstituteUniversity of California–Los Angeles

A multi-institutional, DOE-funded, Bioenergy Research Center dedicated to understanding and modifying Biomass Recalcitrance

322 people in 20 institutions

Somerville et al

Biomass Recalcitrance

• Plant cell walls are complex, heterogenous structures– Three major polymers

• Cellulose, hemicellulose (multiple types), and lignin• Components closely associated and differentially reactive

– Cell wall polysaccharides are structural• More recalcitrant than storage carbohydrates such as starch

– Steps to overcome recalcitrance• Pretreatment- enables efficient enzymatic hydrolysis• Enzymatic hydrolysis is less damaging than pretreatment

• Recalcitrance- genetic and environmental factors– Must screen large populations to find and correlate

underlying principles and effects– Affects pretreatability and enzyme digestibility

Genetics

Process

Conventionalenzyme fermentation

Ethanol

Butanol

Alcohols

Fermentation

Recalcitrance

Consolidatedbioprocessing

Chemicalcatalysis

Hydrocarbons

Syntheticbiology

Hydrocarbons

Environment

Screening Large Populations for Recalcitrance Variants

10s of samples

grams of material

Weeks of analysis

100s/1000s of samples

milligrams of material

Hours of analysis

How is Recalcitrance Affected by:•Environmental variables

•Natural diversity•Genetic manipulation•Enzyme composition

•Pretreatment chemistry•Pretreatment severity

Repeat many times

Fast- many challenges

Slow- huge effort required

Barriers to High Throughput Screening in Biomass Conversion• Defined by large sample sets

– Manual labor in collecting, milling, extracting, starch removal

– Demands small-scale, automated processing

• Biomass is insoluble, heterogeneous– Accurate dispensing, distribution is difficult– Static is major problem

• Severe pretreatment conditions– High temperatures (150-200oC), pressures, low pH

• Solids/Liquids separation– Small scale losses large impact on analysis

• Enzyme systems are complex, slow– Long conversion times (days, not minutes)

• Sugar product detection – HPLC is slow (one plate = 2 days of HPLC time)

• Chemical analysis/composition– Standard techniques too slow

• Systematically addressed/constrained the barriers

100 mi200 km

Skagit (Sedro Woolley)

Skykomish (Monroe)

Puyallup (Orting)Columbia (Longview)

Existing collections (N = 500; 1–2 trees/site)New collections (N = 580; 140–160 trees/site)

Biomass Preparation and Dispensing• Collect and mill by hand• De-starch/extract in “teabags”

– Wash extensively, extract in Soxhlet

• Solids- Symyx MTM Powdernium (2)– Dispense/track quantities

• Automated, 0.1 mg precision– Multiple substrate capacity

• 144 or 200 deck capacity• 15 plate capacity

• Liquid- Beckman-Coulter Biomek FX– Catalyst, enzymes, sugar assays

96-Well Pretreatment Reactor Plate

7

• Single piece reactor plate– Sequential pretreatment/enzymatic saccharification– No Solid-Liquid separation- neutralize only

• Outer dimensions of standard 96-well plate– Robot friendly

• Larger well diameter increases volume capacity• 117 steam/cooling water channels per plate

– 4 channels/well – even heating and cooling

• Ni/Au plated wells add corrosion resistance– Hastalloy, other coatings option for dilute acid

• Stackable to enable multi-plate pretreatments• Custom designed and manufactured

– Available from Aspen Machining, Lakewood, CO, USA

Steam/cooling water

Parr Reactor

Head Plate

Base Plate

Reactor plates

Teflon gasketsAl foil seal

Nuts/bevel washers

Tightening studs

HTP Plate Clamping System Design

Max

imum

20

Plat

esTo

p Pl

ate

Bot

tom

Pla

te

• Stacked system– 20 plates, 1920 pretreatments

• Aluminum head and base plates – Fit snug in Parr reactor – designed for rigidity

• Plate sealing– 0.020” glass-reinforced PFTE

gaskets – Aluminum foil seals on plates

• Steam/cooling water enters through bottom clamping plate

• Even heating well-to-well and vertically

0

20

40

60

80

100

120

140

160

180

200

0 1 2 3

Tem

pera

ture

(o C)

Minutes

Reactor Plate Heating Rate

Well G2

Well D6

Well C11

Closing up Parr Reactor

Steam start

vent closed

y = 0.8382x - 0.0789R² = 0.9958

0

0.5

1

1.5

2

2.5

3

0.0 1.0 2.0 3.0 4.0

Xylo

se b

y H

PLC

(mg/

mL)

Xylose by XDH (mg/mL)

Comparison of Xylose Quantitation Methods

y = 1.0198x + 0.0584R² = 0.9796

0

0.5

1

1.5

2

2.5

3

0.0 0.5 1.0 1.5 2.0 2.5 3.0

Glu

cose

by

HPL

C (m

g/m

L)

Glucose by GOPOD (mg/mL)

Comparison of Glucose Detection Methods

Evaporation Control-Sugar Quantitation• 3 days at 40oC• Clear adhesive seal was inconsistent

– Edge effects• Used magnets in steam ports

– Press fit into aluminum base plate– Thin magnetic stainless steel top plate

Glucose - glucose oxidase/peroxidaseD-Glucose + O2 + H2O D-gluconate + H2O2H2O2 + p-HBA + 4-aminoantipyrine quinoneimine dye (A510)

• GOPOD cross-reactivity– Xylose, arabinose,

xylobiose- none– Galactose, mannose-

trace

Xylose - ß-D-xylose dehydrogenaseß-D-Xylose + NAD+ D-xylonic acid + NADH (A340) + H+

• XDH cross-reactivity– Xylose, arabinose,

mannose- none– Galactose, trace– Xylobiose- moderate

0

20

40

60

80

100

120

0 50 100 150% W

ater

Rem

aini

ng in

Pla

te

Time (h)

Water Loss Throughout IncubationTef lon-coated plates, 40oC, static

1. Plate + seal only2. Plate + Seal + 13 magnets and steel plates

Parallel Plate System Overview

10

Stack Plates

Pretreat

Load PlatesAdd Catalyst and

Seal

Disassemble, add enzymes, buffer

Seal,Incubate

Enzyme Mediated Sugar Detection

Robotics SystemsEnable High Volume

Custom Plates and Reactor

Analysis of Cell Wall Chemistry• MBMS- rapid estimate of cell wall chemistry

– High information content

– Complements wet chemistry

• PCA of MS from ~1500 greenhouse poplar. – Black = low nitrogen

– Purple = high nitrogen.

• The two clusters indicate differing cell wall chemistry – Different lignin composition

11

-8

-6

-4

-2

0

2

4

6

-8 -6 -4 -2 0 2 4 6 8 10 12 RESULT1, X-expl: 62%,13%

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UF343-5-CHEM

UF343-6-CHEM

UF343-6-CHEM

UF344-1-CHEM

UF344-1-CHEMUF344-2-CHEM

UF344-2-CHEM

UF344-5-CHEM

UF344-5-CHEM

UF344-6-CHEM

UF344-6-CHEM

UF345-1-CHEMUF345-1-CHEM

UF345-2-CHEMUF345-2-CHEM

UF345-5-CHEM

UF345-5-CHEM

UF35-1-CHEM

UF35-1-CHEM

UF35-2-CHEM

UF35-5-CHEMUF35-5-CHEMUF35-6-CHEMUF35-6-CHEM

UF352-1-CHEM

UF352-1-CHEM

UF352-2-CHEM

UF352-2-CHEM

UF352-5-CHEMUF352-5-CHEM

UF352-6-CHEM

UF352-6-CHEM

UF359-1-CHEM

UF359-1-CHEMUF359-2-CHEM

UF359-2-CHEM

UF359-5-CHEM

UF359-5-CHEM

UF359-6-CHEM

UF359-6-CHEM

UF36-1-CHEMUF36-1-CHEM

UF36-2-CHEMUF36-2-CHEM

UF36-5-CHEM

UF36-5-CHEM

UF36-6-CHEMUF36-6-CHEM

UF361-1-CHEM

UF361-1-CHEMUF361-2-CHEM

UF361-2-CHEM

UF361-5-CHEM

UF361-5-CHEM

UF361-6-CHEMUF361-6-CHEM

UF366-1-CHEM

UF366-1-CHEM

UF366-1-CHEM

UF366-2-CHEMUF366-2-CHEM

UF366-5-CHEM

UF366-5-CHEMUF366-6-CHEM

UF366-6-CHEM

UF368-1-CHEM

UF368-1-CHEMUF368-1-CHEM

UF368-2-CHEMUF368-2-CHEM

UF368-5-CHEM

UF368-5-CHEM

UF368-6-CHEM

UF368-6-CHEM

UF370-1-CHEM

UF370-1-CHEMUF370-2-CHEM

UF370-2-CHEM

UF370-5-CHEM

UF370-5-CHEM

UF370-6-CHEM

UF370-6-CHEM

UF372-1-CHEM

UF372-1-CHEM

UF372-2-CHEM

UF372-2-CHEM

UF372-5-CHEM

UF372-5-CHEM

UF372-6-CHEM

UF372-6-CHEMUF38-2-CHEMUF38-2-CHEM

UF38-5-CHEM

UF38-5-CHEM

UF38-6-CHEM

UF38-6-CHEM

UF380-1-CHEM

UF380-1-CHEM

UF380-2-CHEMUF380-2-CHEM

UF380-5-CHEM

UF380-5-CHEM

UF380-6-CHEMUF380-6-CHEM

UF381-1-CHEM

UF381-1-CHEM

UF381-2-CHEMUF381-2-CHEM

UF381-5-CHEM

UF381-5-CHEM

UF381-6-CHEM

UF381-6-CHEM

UF382-1-CHEM

UF382-1-CHEM

UF382-2-CHEM

UF382-2-CHEM

UF382-5-CHEM

UF382-5-CHEM

UF382-6-CHEMUF382-6-CHEM

UF383-1-CHEM

UF383-1-CHEM

UF383-2-CHEM

UF383-2-CHEM

UF383-5-CHEM

UF383-5-CHEMUF383-6-CHEMUF383-6-CHEM

UF385-1-CHEM

UF385-1-CHEM

UF385-2-CHEMUF385-2-CHEM

UF385-5-CHEM

UF385-5-CHEM

UF385-6-CHEMUF385-6-CHEM

UF389-1-CHEM

UF389-1-CHEMUF389-1-CHEM

UF389-2-CHEMUF389-2-CHEM

UF389-5-CHEM

UF389-5-CHEMUF389-6-CHEM

UF389-6-CHEM

UF39-1-CHEM

UF39-1-CHEMUF39-2-CHEM

UF39-5-CHEM

UF39-5-CHEMUF39-6-CHEMUF39-6-CHEM

UF390-1-CHEM

UF390-1-CHEMUF390-2-CHEM

UF390-2-CHEM

UF392-1-CHEMUF392-1-CHEM

UF392-2-CHEMUF392-2-CHEM

UF392-5-CHEM

UF392-5-CHEMUF392-6-CHEMUF392-6-CHEM

UF394-1-CHEM

UF394-1-CHEM

UF394-2-CHEMUF394-2-CHEM

UF394-5-CHEM

UF394-5-CHEM

UF394-6-CHEM

UF394-6-CHEM

UF395-1-CHEM

UF395-1-CHEM

UF395-5-CHEM

UF395-5-CHEM

UF395-6-CHEM

UF396-1-CHEM

UF396-1-CHEM

UF396-2-CHEM

UF396-2-CHEM

UF396-5-CHEM

UF396-5-CHEM

UF396-6-CHEMUF396-6-CHEM

UF397-1-CHEM

UF397-1-CHEM

UF397-1-CHEM

UF397-2-CHEMUF397-2-CHEM

UF397-5-CHEM

UF397-5-CHEM

UF397-6-CHEMUF397-6-CHEM

UF398-1-CHEM

UF398-1-CHEM

UF398-2-CHEM

UF398-2-CHEM

UF398-5-CHEM

UF398-5-CHEMUF398-6-CHEMUF398-6-CHEM

UF40-1-CHEM

UF40-1-CHEM

UF40-2-CHEMUF40-2-CHEM

UF40-5-CHEM

UF40-5-CHEM

UF40-6-CHEM

UF40-6-CHEM

UF401-1-CHEM

UF401-1-CHEM

UF401-2-CHEM

UF401-2-CHEM

UF401-5-CHEM

UF401-5-CHEM

UF402-1-CHEMUF402-1-CHEM

UF402-2-CHEMUF402-2-CHEM

UF402-5-CHEM

UF402-5-CHEM

UF402-6-CHEMUF402-6-CHEM

UF404-1-CHEM

UF404-1-CHEMUF404-2-CHEM

UF404-2-CHEM

UF404-5-CHEM

UF404-5-CHEMUF404-6-CHEMUF404-6-CHEM

UF405-1-CHEM

UF405-1-CHEMUF405-1-CHEM

UF405-2-CHEMUF405-2-CHEM

UF405-5-CHEM

UF405-5-CHEM

UF405-6-CHEMUF405-6-CHEM

UF406-1-CHEM

UF406-1-CHEM

UF406-2-CHEMUF406-2-CHEM

UF406-5-CHEM

UF406-5-CHEMUF406-6-CHEMUF406-6-CHEM

UF407-1-CHEM

UF407-1-CHEMUF407-2-CHEMUF407-2-CHEM

UF407-5-CHEM

UF407-5-CHEMUF407-6-CHEM

UF407-6-CHEM

UF408-1-CHEM

UF408-1-CHEM

UF408-2-CHEMUF408-2-CHEM

UF408-5-CHEM

UF408-5-CHEM

UF408-6-CHEM

UF408-6-CHEM

UF409-1-CHEMUF409-1-CHEM

UF409-2-CHEMUF409-2-CHEM

UF409-5-CHEM

UF409-5-CHEM

UF409-6-CHEM

UF409-6-CHEM

UF417-1-CHEM

UF417-1-CHEM

UF417-2-CHEM

UF417-2-CHEM

UF417-5-CHEM

UF417-5-CHEM

UF417-6-CHEMUF417-6-CHEM

UF418-1-CHEM

UF418-1-CHEMUF418-1-CHEM

UF418-2-CHEM

UF418-2-CHEM

UF418-5-CHEM

UF418-5-CHEM

UF418-6-CHEMUF418-6-CHEM

UF419-1-CHEM

UF419-1-CHEM

UF419-2-CHEMUF419-2-CHEM

UF419-5-CHEM

UF419-5-CHEM

UF419-6-CHEM

UF419-6-CHEM

UF42-1-CHEM

UF42-1-CHEM

UF42-2-CHEMUF42-2-CHEM

UF42-5-CHEM

UF42-5-CHEMUF42-6-CHEM

UF42-6-CHEM

UF421-5-CHEM

UF421-5-CHEMUF421-6-CHEMUF421-6-CHEM

UF435-1-CHEM

UF435-1-CHEMUF435-1-CHEMUF435-2-CHEM

UF435-5-CHEM

UF435-5-CHEMUF435-6-CHEM

UF435-6-CHEM

UF436-1-CHEM

UF436-1-CHEM

UF436-2-CHEMUF436-2-CHEM

UF436-5-CHEM

UF436-5-CHEM

UF436-6-CHEMUF436-6-CHEM

UF437-2-CHEM

UF437-2-CHEM

UF437-5-CHEM

UF437-5-CHEMUF437-6-CHEM

UF440-1-CHEM

UF440-1-CHEM

UF440-2-CHEMUF440-2-CHEM

UF440-5-CHEM

UF440-5-CHEMUF440-6-CHEM

UF441-1-CHEM

UF441-1-CHEM

UF441-2-CHEMUF441-2-CHEM

UF441-5-CHEM

UF441-5-CHEM

UF441-6-CHEM

UF441-6-CHEMUF445-1-CHEM

UF445-1-CHEM

UF445-1-CHEM

UF445-2-CHEM

UF445-2-CHEM

UF445-5-CHEM

UF445-5-CHEM

UF445-6-CHEMUF445-6-CHEM

UF451-1-CHEM

UF451-1-CHEMUF451-2-CHEMUF451-2-CHEM

UF451-5-CHEM

UF451-5-CHEM

UF451-6-CHEM

UF452-1-CHEMUF452-1-CHEM

UF452-2-CHEM

UF452-2-CHEM

UF452-5-CHEM

UF452-5-CHEM

UF452-6-CHEMUF452-6-CHEM

UF453-1-CHEM

UF453-1-CHEMUF453-2-CHEMUF453-2-CHEM

UF453-5-CHEM

UF453-5-CHEMUF453-6-CHEM

UF453-6-CHEMUF456-1-CHEM

UF456-1-CHEMUF456-2-CHEMUF456-2-CHEM

UF456-5-CHEM

UF456-5-CHEM

UF456-6-CHEM

UF457-1-CHEMUF457-1-CHEM

UF457-2-CHEM

UF457-2-CHEM

UF457-5-CHEM

UF457-5-CHEM

UF457-6-CHEMUF457-6-CHEM

UF46-1-CHEM

UF46-1-CHEM

UF46-2-CHEM

UF46-5-CHEM

UF46-5-CHEM

UF46-6-CHEM

UF46-6-CHEM

UF460-2-CHEMUF460-2-CHEM

UF460-5-CHEM

UF460-5-CHEMUF460-6-CHEM

UF460-6-CHEM

UF461-1-CHEM

UF461-1-CHEM

UF461-2-CHEM

UF461-2-CHEM

UF461-5-CHEM

UF461-5-CHEMUF461-6-CHEMUF461-6-CHEM

UF464-1-CHEM

UF464-1-CHEM

UF464-2-CHEM

UF464-2-CHEM

UF464-5-CHEM

UF464-5-CHEM

UF464-6-CHEM

UF464-6-CHEM

UF465-1-CHEM

UF465-1-CHEM

UF465-2-CHEMUF465-2-CHEM

UF465-5-CHEM

UF465-5-CHEM

UF465-6-CHEMUF465-6-CHEM

UF467-1-CHEM

UF467-1-CHEM

UF467-2-CHEM

UF467-2-CHEM

UF467-5-CHEM

UF467-5-CHEMUF467-6-CHEM

UF467-6-CHEM

UF468-1-CHEM

UF468-1-CHEM

UF468-2-CHEM

UF468-2-CHEM

UF468-5-CHEM

UF468-5-CHEM

UF468-6-CHEMUF468-6-CHEM

UF47-1-CHEM

UF47-1-CHEM

UF47-1-CHEM

UF47-2-CHEM

UF47-2-CHEM

UF473-2-CHEMUF473-2-CHEM

UF473-5-CHEM

UF473-5-CHEM

UF473-6-CHEMUF473-6-CHEM

UF476-5-CHEMUF476-5-CHEM

UF476-6-CHEMUF476-6-CHEM

UF477-1-CHEM

UF477-1-CHEM

UF477-2-CHEMUF477-2-CHEM

UF477-5-CHEM

UF477-5-CHEM

UF477-6-CHEM

UF477-6-CHEM

UF479-2-CHEMUF479-2-CHEM

UF479-5-CHEM

UF479-5-CHEM

UF479-6-CHEM

UF479-6-CHEM

UF481-1-CHEM

UF481-1-CHEM

UF481-2-CHEMUF481-2-CHEM

UF481-5-CHEM

UF481-5-CHEMUF481-6-CHEM

UF481-6-CHEM

UF488-1-CHEM

UF488-1-CHEMUF488-2-CHEMUF488-2-CHEMUF488-5-CHEM

UF488-5-CHEM

UF488-6-CHEM

UF488-6-CHEM

UF489-1-CHEMUF489-1-CHEMUF489-2-CHEM

UF489-2-CHEM

UF489-6-CHEMUF489-6-CHEM

UF49-1-CHEM

UF49-1-CHEM

UF49-2-CHEMUF49-2-CHEM

UF49-5-CHEM

UF49-5-CHEM

UF49-6-CHEMUF49-6-CHEMUF490-1-CHEM

UF490-1-CHEM

UF490-2-CHEMUF490-2-CHEM

UF490-5-CHEM

UF490-5-CHEM

UF490-6-CHEMUF490-6-CHEM

UF491-1-CHEM

UF491-1-CHEM

UF491-2-CHEMUF491-2-CHEM

UF491-5-CHEM

UF491-5-CHEM

UF491-6-CHEMUF491-6-CHEM

UF492-1-CHEMUF492-1-CHEM

UF492-2-CHEM

UF492-2-CHEM

UF492-5-CHEM

UF492-5-CHEMUF492-6-CHEM

UF492-6-CHEM

UF493-1-CHEM

UF493-1-CHEM

UF493-2-CHEMUF493-2-CHEM

UF493-5-CHEM

UF493-5-CHEM

UF493-6-CHEMUF493-6-CHEM

UF494-1-CHEM

UF494-1-CHEM

UF494-1-CHEM

UF494-2-CHEMUF494-2-CHEM

UF494-5-CHEM

UF494-5-CHEM

UF494-6-CHEM

UF494-6-CHEM

UF497-1-CHEM

UF497-1-CHEM

UF497-2-CHEMUF497-2-CHEM

UF497-5-CHEMUF497-5-CHEM

UF497-6-CHEM

UF497-6-CHEM

UF498-1-CHEM

UF498-1-CHEMUF498-1-CHEM

UF498-2-CHEM

UF498-2-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-5-CHEM

UF498-6-CHEMUF498-6-CHEM

UF499-1-CHEM

UF499-1-CHEM

UF499-2-CHEMUF499-2-CHEM

UF499-5-CHEM

UF499-5-CHEMUF499-6-CHEM

UF499-6-CHEM

UF500-1-CHEM

UF500-1-CHEM

UF500-2-CHEMUF500-2-CHEM

UF500-5-CHEM

UF500-5-CHEM

UF500-6-CHEM

UF500-6-CHEM

UF503-1-CHEM

UF503-1-CHEM

UF503-2-CHEM

UF503-2-CHEM

UF503-6-CHEMUF503-6-CHEM

UF505-1-CHEM

UF505-1-CHEM

UF505-2-CHEMUF505-2-CHEM

UF505-5-CHEM

UF505-5-CHEM

UF505-6-CHEM

UF505-6-CHEM

UF512-1-CHEM

UF512-1-CHEM

UF512-5-CHEM

UF512-5-CHEMUF512-6-CHEMUF512-6-CHEM

UF513-1-CHEM

UF513-1-CHEM

UF513-2-CHEM

UF513-2-CHEM

UF513-5-CHEM

UF513-5-CHEM

UF513-6-CHEMUF513-6-CHEM

UF515-1-CHEM

UF515-1-CHEMUF515-5-CHEM

UF515-5-CHEM

UF515-6-CHEMUF515-6-CHEM

UF516-1-CHEM

UF516-1-CHEM

UF516-2-CHEM

UF516-2-CHEM

UF516-5-CHEM

UF516-5-CHEMUF516-6-CHEMUF516-6-CHEM

UF518-1-CHEM

UF518-1-CHEM

UF518-2-CHEM

UF518-2-CHEM

UF518-5-CHEM

UF518-5-CHEMUF518-6-CHEM

UF518-6-CHEM

UF525-1-CHEM

UF525-1-CHEM

UF525-1-CHEM

UF525-2-CHEM

UF525-2-CHEM

UF526-1-CHEM

UF526-1-CHEMUF526-2-CHEM

UF526-2-CHEM

UF526-5-CHEM

UF526-5-CHEM

UF526-6-CHEM

UF526-6-CHEM

UF529-1-CHEM

UF529-1-CHEM

UF529-2-CHEM

UF529-2-CHEM

UF529-5-CHEM

UF529-5-CHEM

UF529-6-CHEMUF529-6-CHEM

UF53-1-CHEM

UF53-1-CHEM

UF53-2-CHEMUF53-2-CHEM

UF53-5-CHEM

UF53-5-CHEM

UF53-6-CHEMUF53-6-CHEM

UF532-1-CHEM

UF532-1-CHEMUF532-2-CHEM

UF532-5-CHEM

UF532-5-CHEMUF532-6-CHEM

UF532-6-CHEM

UF533-1-CHEMUF533-1-CHEM

UF533-2-CHEM

UF533-2-CHEM

UF533-5-CHEM

UF533-5-CHEM

UF533-6-CHEMUF533-6-CHEM

UF534-1-CHEM

UF534-1-CHEM

UF534-2-CHEM

UF534-2-CHEM

UF534-5-CHEM

UF534-5-CHEMUF534-6-CHEM

UF535-1-CHEM

UF535-1-CHEM

UF535-2-CHEM

UF535-2-CHEM

UF535-5-CHEM

UF535-5-CHEM

UF535-6-CHEMUF535-6-CHEM

UF537-1-CHEM

UF537-1-CHEM

UF537-2-CHEMUF537-2-CHEM

UF537-5-CHEM

UF537-5-CHEM

UF537-6-CHEM

UF537-6-CHEM

UF538-1-CHEM

UF538-1-CHEMUF538-2-CHEM

UF538-2-CHEM

UF538-5-CHEM

UF538-5-CHEM

UF538-6-CHEMUF538-6-CHEM

UF539-1-CHEM

UF539-1-CHEM

UF539-2-CHEMUF539-2-CHEM

UF539-5-CHEM

UF539-5-CHEM

UF539-6-CHEMUF539-6-CHEM

UF54-1-CHEM

UF54-1-CHEM

UF54-2-CHEM

UF54-5-CHEM

UF54-5-CHEMUF54-6-CHEMUF54-6-CHEM

UF541-1-CHEM

UF541-1-CHEM

UF541-2-CHEM

UF541-2-CHEM

UF541-5-CHEM

UF541-5-CHEM

UF541-6-CHEMUF541-6-CHEM

UF542-1-CHEMUF542-1-CHEM

UF542-2-CHEM

UF542-5-CHEM

UF542-5-CHEMUF542-6-CHEM

UF542-6-CHEM

UF543-2-CHEM

UF543-2-CHEM

UF544-1-CHEM

UF544-1-CHEM

UF544-2-CHEMUF544-2-CHEM

UF544-5-CHEM

UF544-5-CHEM

UF544-6-CHEMUF544-6-CHEM

UF546-1-CHEM

UF546-1-CHEM

UF546-2-CHEMUF546-2-CHEM

UF546-5-CHEM

UF546-5-CHEM

UF546-6-CHEM

UF546-6-CHEM

UF547-1-CHEM

UF547-1-CHEM

UF547-2-CHEM

UF547-5-CHEM

UF547-5-CHEM

UF547-6-CHEM

UF552-1-CHEMUF552-1-CHEM

UF552-2-CHEM

UF552-2-CHEM

UF552-5-CHEMUF552-5-CHEM

UF552-6-CHEM

UF552-6-CHEM

UF554-1-CHEM

UF554-1-CHEM

UF554-2-CHEMUF554-2-CHEM

UF554-5-CHEM

UF554-5-CHEM

UF554-6-CHEM

UF554-6-CHEM

UF56-1-CHEM

UF56-1-CHEM

UF56-2-CHEM

UF56-2-CHEM

UF56-5-CHEM

UF56-5-CHEM

UF56-6-CHEMUF56-6-CHEM

UF561-1-CHEM

UF561-1-CHEM

UF561-2-CHEMUF561-2-CHEM

UF561-5-CHEMUF561-5-CHEM

UF561-6-CHEMUF561-6-CHEM UF562-1-CHEM

UF562-1-CHEM

UF562-2-CHEMUF562-2-CHEM

UF562-5-CHEM

UF562-5-CHEM

UF562-6-CHEMUF562-6-CHEM

UF565-1-CHEMUF565-1-CHEM

UF565-2-CHEMUF565-2-CHEM

UF565-5-CHEM

UF565-5-CHEM

UF565-6-CHEM

UF565-6-CHEM

UF567-1-CHEM

UF567-1-CHEM

UF567-2-CHEMUF567-2-CHEM

UF567-5-CHEM

UF567-5-CHEM

UF567-6-CHEMUF567-6-CHEM

UF569-2-CHEMUF569-2-CHEM

UF569-5-CHEM

UF569-5-CHEM

UF569-6-CHEMUF569-6-CHEM UF570-1-CHEM

UF570-1-CHEM

UF570-2-CHEM

UF570-2-CHEM

UF570-5-CHEM

UF570-5-CHEM

UF570-6-CHEM

UF570-6-CHEM

UF571-1-CHEM

UF571-1-CHEM

UF571-1-CHEM

UF571-2-CHEM

UF571-2-CHEM

UF572-2-CHEM

UF572-2-CHEM

UF572-5-CHEMUF572-5-CHEM

UF572-6-CHEMUF572-6-CHEM

UF576-1-CHEM

UF576-1-CHEM

UF576-2-CHEMUF576-2-CHEM

UF576-5-CHEM

UF576-5-CHEMUF576-6-CHEM

UF576-6-CHEM

UF578-5-CHEM

UF578-5-CHEM

UF578-6-CHEMUF578-6-CHEM UF579-1-CHEM

UF579-1-CHEM

UF579-2-CHEM

UF579-2-CHEM

UF579-5-CHEM

UF579-5-CHEM

UF58-1-CHEMUF58-1-CHEM

UF58-2-CHEM

UF58-2-CHEM

UF58-5-CHEM

UF58-5-CHEM

UF58-6-CHEM

UF58-6-CHEM

UF581-1-CHEM

UF581-1-CHEM

UF581-2-CHEM

UF581-2-CHEM

UF581-5-CHEM

UF581-5-CHEM

UF581-6-CHEM

UF581-6-CHEM

UF582-1-CHEM

UF582-1-CHEM

UF582-2-CHEM

UF582-2-CHEM

UF582-5-CHEM

UF582-5-CHEM

UF582-6-CHEM

UF582-6-CHEM

UF584-1-CHEM

UF584-1-CHEM

UF584-2-CHEM

UF584-2-CHEM

UF584-5-CHEM

UF584-5-CHEMUF584-6-CHEMUF584-6-CHEM

UF585-1-CHEM

UF585-1-CHEM

UF585-2-CHEM

UF585-2-CHEM

UF585-5-CHEM

UF585-5-CHEMUF585-6-CHEMUF585-6-CHEM

UF586-1-CHEM

UF586-1-CHEM

UF586-2-CHEMUF586-2-CHEM

UF586-5-CHEM

UF586-5-CHEMUF586-6-CHEM

UF586-6-CHEM

UF587-1-CHEM

UF587-1-CHEM

UF587-2-CHEMUF587-2-CHEM

UF587-5-CHEM

UF587-5-CHEM

UF587-6-CHEM

UF587-6-CHEM

UF588-1-CHEM

UF588-1-CHEM

UF588-2-CHEMUF588-2-CHEM

UF588-5-CHEM

UF588-5-CHEM

UF588-6-CHEMUF588-6-CHEM

UF589-1-CHEM

UF589-1-CHEM

UF589-2-CHEM

UF589-2-CHEM

UF589-5-CHEM

UF589-5-CHEM

UF589-6-CHEM

UF59-1-CHEM

UF59-1-CHEMUF59-2-CHEM

UF59-2-CHEM

UF59-5-CHEM

UF59-5-CHEM

UF59-6-CHEMUF59-6-CHEM

UF590-1-CHEM

UF590-1-CHEM

UF590-2-CHEM

UF590-2-CHEM

UF590-5-CHEMUF590-5-CHEM

UF590-6-CHEMUF590-6-CHEM

UF596-2-CHEMUF596-2-CHEM

UF596-5-CHEM

UF596-5-CHEM

UF596-6-CHEMUF596-6-CHEM

UF600-1-CHEM

UF600-1-CHEM

UF600-1-CHEMUF600-2-CHEM

UF600-2-CHEM

UF600-5-CHEM

UF600-5-CHEM

UF600-6-CHEMUF600-6-CHEM

UF61-1-CHEM

UF61-1-CHEM

UF61-2-CHEM

UF61-2-CHEM

UF61-5-CHEM

UF61-5-CHEM

UF61-6-CHEM

UF61-6-CHEM

UF62-1-CHEM

UF62-1-CHEM

UF62-2-CHEMUF62-2-CHEM

UF62-5-CHEM

UF62-5-CHEM

UF62-6-CHEMUF62-6-CHEM

UF65-1-CHEMUF65-1-CHEM

UF65-2-CHEMUF65-2-CHEM

UF66-1-CHEM

UF66-1-CHEM

UF66-2-CHEMUF66-2-CHEMUF66-5-CHEM

UF66-5-CHEM

UF66-6-CHEMUF66-6-CHEM

UF68-1-CHEMUF68-1-CHEM

UF68-2-CHEM

UF68-2-CHEM

UF68-5-CHEM

UF68-5-CHEM

UF68-6-CHEMUF68-6-CHEM

UF69-1-CHEM

UF69-1-CHEM

UF69-2-CHEM

UF69-2-CHEM

UF69-5-CHEM

UF69-5-CHEM

UF69-6-CHEMUF69-6-CHEM

UF7-1-CHEM

UF7-1-CHEMUF7-2-CHEMUF7-2-CHEM

UF7-5-CHEM

UF7-5-CHEM

UF7-6-CHEMUF7-6-CHEM

UF70-1-CHEMUF70-1-CHEM

UF70-2-CHEMUF70-2-CHEM

UF70-5-CHEM

UF70-5-CHEM

UF70-6-CHEM

UF70-6-CHEM

UF700-1-CHEM

UF700-1-CHEM

UF700-2-CHEMUF700-2-CHEM

UF700-5-CHEM

UF700-5-CHEMUF700-6-CHEM

UF700-6-CHEM

UF701-1-CHEMUF701-1-CHEM

UF701-2-CHEMUF701-2-CHEM

UF701-5-CHEM

UF701-5-CHEM

UF701-6-CHEMUF701-6-CHEM

UF703-1-CHEM

UF703-1-CHEMUF703-2-CHEM

UF703-2-CHEM

UF703-5-CHEM

UF703-5-CHEM

UF703-6-CHEMUF703-6-CHEM

UF705-1-CHEM

UF705-1-CHEMUF705-1-CHEMUF705-2-CHEM

UF705-2-CHEM

UF705-5-CHEM

UF705-5-CHEMUF705-6-CHEM

UF705-6-CHEMUF706-1-CHEM

UF706-1-CHEM

UF706-2-CHEM

UF706-2-CHEM

UF706-5-CHEM

UF706-5-CHEM

UF706-6-CHEMUF706-6-CHEM

UF707-1-CHEM

UF707-1-CHEMUF707-2-CHEMUF707-2-CHEM

UF707-5-CHEM

UF707-5-CHEM

UF707-6-CHEM

UF707-6-CHEM

UF709-1-CHEM

UF709-1-CHEM

UF709-2-CHEM

UF709-2-CHEM

UF709-5-CHEM

UF709-5-CHEMUF709-6-CHEM

UF709-6-CHEM

UF710-1-CHEM

UF710-1-CHEM

UF710-1-CHEM

UF710-2-CHEMUF710-2-CHEM

UF710-5-CHEM

UF710-5-CHEM

UF710-6-CHEMUF710-6-CHEM

UF714-1-CHEM

UF714-1-CHEM

UF714-1-CHEM

UF714-2-CHEM

UF714-2-CHEM

UF714-5-CHEM

UF714-5-CHEMUF714-6-CHEM

UF714-6-CHEM

UF715-1-CHEM

UF715-1-CHEM

UF715-1-CHEM

UF715-2-CHEMUF715-2-CHEM

UF715-5-CHEM

UF715-5-CHEM

UF715-6-CHEMUF715-6-CHEM

UF716-1-CHEM

UF716-1-CHEM

UF716-2-CHEMUF716-2-CHEM

UF716-5-CHEM

UF716-5-CHEM

UF716-6-CHEMUF716-6-CHEM

UF717-1-CHEM

UF717-1-CHEM

UF717-2-CHEMUF717-2-CHEM

UF717-5-CHEM

UF717-5-CHEM

UF717-6-CHEM

UF717-6-CHEM

UF718-1-CHEM

UF718-1-CHEM

UF718-2-CHEM

UF718-2-CHEM

UF718-5-CHEM

UF718-5-CHEM

UF718-6-CHEM

UF718-6-CHEM

UF720-1-CHEM

UF720-1-CHEM

UF720-2-CHEMUF720-2-CHEM

UF720-5-CHEM

UF720-5-CHEM

UF720-6-CHEM

UF721-1-CHEM

UF721-1-CHEM

UF721-2-CHEM

UF721-2-CHEM

UF722-1-CHEM

UF722-1-CHEM

UF722-2-CHEM

UF722-2-CHEM

UF722-5-CHEM

UF722-5-CHEM

UF722-6-CHEM

UF722-6-CHEM

UF723-1-CHEM

UF723-1-CHEM

UF723-2-CHEMUF723-2-CHEM

UF723-5-CHEM

UF723-5-CHEM

UF723-6-CHEMUF723-6-CHEM

UF724-1-CHEM

UF724-1-CHEMUF724-2-CHEMUF724-2-CHEM

UF724-5-CHEM

UF724-5-CHEM

UF724-6-CHEM

UF727-1-CHEM

UF727-1-CHEM

UF727-2-CHEMUF727-2-CHEM

UF727-5-CHEM

UF727-5-CHEM

UF727-6-CHEM

UF727-6-CHEM

UF728-5-CHEM

UF728-5-CHEM

UF728-6-CHEM

UF729-1-CHEM

UF729-1-CHEM

UF729-2-CHEM

UF729-2-CHEM

UF729-5-CHEM

UF729-5-CHEM

UF729-6-CHEM

UF729-6-CHEM

UF73-1-CHEM

UF73-1-CHEM

UF73-2-CHEM

UF73-2-CHEM UF73-5-CHEM

UF73-5-CHEMUF73-6-CHEMUF73-6-CHEM

UF731-1-CHEMUF731-1-CHEM

UF731-2-CHEM

UF731-2-CHEM

UF731-5-CHEM

UF731-5-CHEMUF731-6-CHEM

UF731-6-CHEM

UF734-1-CHEM

UF734-1-CHEMUF734-2-CHEM

UF734-2-CHEM

UF734-5-CHEM

UF734-5-CHEM

UF734-6-CHEMUF734-6-CHEM

UF735-1-CHEM

UF735-1-CHEM

UF735-2-CHEMUF735-2-CHEM

UF735-5-CHEM

UF735-5-CHEM

UF735-6-CHEMUF735-6-CHEM

UF74-1-CHEM

UF74-1-CHEM

UF74-2-CHEMUF74-2-CHEM

UF74-5-CHEM

UF74-5-CHEM

UF74-6-CHEMUF74-6-CHEM UF741-1-CHEM

UF741-1-CHEMUF741-2-CHEM

UF741-2-CHEM

UF743-1-CHEM

UF743-1-CHEM

UF743-2-CHEM

UF743-2-CHEM

UF743-5-CHEM

UF743-5-CHEM

UF743-6-CHEM

UF743-6-CHEM

UF745-1-CHEM

UF745-1-CHEMUF745-2-CHEMUF745-2-CHEM

UF745-5-CHEM

UF745-5-CHEM

UF745-6-CHEMUF745-6-CHEM

UF748-1-CHEM

UF748-1-CHEM

UF748-2-CHEM

UF748-2-CHEM

UF748-5-CHEM

UF748-5-CHEM

UF748-6-CHEM

UF748-6-CHEM

UF751-1-CHEM

UF751-1-CHEMUF751-2-CHEM

UF751-2-CHEM UF751-5-CHEMUF751-5-CHEM

UF751-6-CHEMUF751-6-CHEM

UF754-1-CHEM

UF754-1-CHEM

UF754-2-CHEM

UF754-2-CHEM

UF754-5-CHEMUF754-5-CHEM

UF754-6-CHEM

UF754-6-CHEMUF755-1-CHEM

UF755-1-CHEM

UF755-2-CHEMUF755-2-CHEM

UF755-5-CHEMUF755-5-CHEM

UF755-6-CHEM

UF78-1-CHEM

UF78-1-CHEM

UF78-2-CHEM

UF78-2-CHEM

UF78-5-CHEM

UF78-5-CHEMUF78-6-CHEM

UF78-6-CHEM

UF79-1-CHEM

UF79-1-CHEM

UF79-2-CHEM

UF79-2-CHEM

UF79-5-CHEM

UF79-5-CHEMUF79-6-CHEMUF79-6-CHEM

UF800-1-CHEMUF800-1-CHEM

UF800-2-CHEM

UF800-2-CHEM

UF800-5-CHEM

UF800-5-CHEM

UF800-6-CHEMUF800-6-CHEM

UF801-1-CHEM

UF801-1-CHEMUF801-2-CHEM

UF801-2-CHEM

UF801-5-CHEM

UF801-5-CHEM

UF801-6-CHEMUF801-6-CHEM

UF802-1-CHEMUF802-1-CHEM

UF802-2-CHEMUF802-2-CHEMUF802-5-CHEM

UF802-5-CHEM

UF802-6-CHEM

UF802-6-CHEM

UF803-1-CHEM

UF803-1-CHEM

UF803-2-CHEM

UF803-2-CHEM

UF803-5-CHEM

UF803-5-CHEM

UF803-6-CHEM

UF803-6-CHEM

UF804-1-CHEM

UF804-1-CHEM

UF804-2-CHEM

UF804-2-CHEM

UF804-5-CHEM

UF804-5-CHEM

UF804-6-CHEMUF804-6-CHEM

UF805-1-CHEM

UF805-1-CHEM

UF805-2-CHEMUF805-2-CHEM

UF805-5-CHEM

UF805-5-CHEM

UF805-6-CHEM

UF805-6-CHEM

UF806-1-CHEM

UF806-1-CHEMUF806-2-CHEMUF806-5-CHEM

UF806-5-CHEM

UF806-6-CHEM

UF806-6-CHEMUF807-1-CHEM

UF807-1-CHEM

UF807-2-CHEMUF807-2-CHEM

UF807-5-CHEM

UF807-5-CHEMUF807-6-CHEMUF807-6-CHEM

UF808-1-CHEM

UF808-1-CHEMUF808-1-CHEM

UF808-2-CHEMUF808-2-CHEM UF808-5-CHEM

UF808-5-CHEM

UF808-6-CHEM

UF808-6-CHEM

UF809-1-CHEM

UF809-1-CHEM

UF809-2-CHEM

UF809-5-CHEM

UF809-5-CHEM

UF809-6-CHEM

UF809-6-CHEM

UF810-1-CHEM

UF810-1-CHEM

UF810-5-CHEM

UF810-5-CHEM

UF810-6-CHEMUF810-6-CHEM

UF811-1-CHEM

UF811-1-CHEM

UF811-2-CHEMUF811-2-CHEM UF812-1-CHEM

UF812-1-CHEM

UF812-2-CHEM

UF812-2-CHEM

UF812-5-CHEM

UF812-5-CHEM

UF812-6-CHEM

UF813-1-CHEM

UF813-1-CHEMUF813-2-CHEMUF813-2-CHEM

UF813-5-CHEM

UF813-5-CHEM

UF813-6-CHEMUF813-6-CHEM

UF814-1-CHEM

UF814-1-CHEMUF814-2-CHEM

UF814-2-CHEM

UF814-5-CHEM

UF814-5-CHEM

UF814-6-CHEM

UF814-6-CHEM

UF815-1-CHEM

UF815-1-CHEM

UF815-2-CHEM

UF815-2-CHEM

UF815-5-CHEM

UF815-5-CHEM

UF815-6-CHEMUF815-6-CHEM

UF816-1-CHEM

UF816-1-CHEM

UF816-2-CHEMUF816-2-CHEM

UF816-5-CHEM

UF816-5-CHEM

UF816-6-CHEM

UF816-6-CHEM

UF817-5-CHEM

UF817-5-CHEM

UF817-6-CHEMUF817-6-CHEM

UF818-1-CHEM

UF818-1-CHEM

UF818-2-CHEMUF818-2-CHEM

UF818-5-CHEM

UF818-5-CHEM

UF818-6-CHEMUF818-6-CHEM

UF819-1-CHEM

UF819-1-CHEM

UF819-2-CHEMUF819-2-CHEM

UF819-5-CHEM

UF819-5-CHEM

UF819-6-CHEMUF819-6-CHEM

UF820-1-CHEM

UF820-1-CHEM

UF820-2-CHEM

UF820-2-CHEM

UF820-5-CHEM

UF820-5-CHEM

UF820-6-CHEMUF820-6-CHEM

UF821-2-CHEMUF821-2-CHEM

UF821-5-CHEM

UF821-5-CHEM

UF821-6-CHEMUF821-6-CHEM

UF822-1-CHEM

UF822-1-CHEM

UF822-2-CHEMUF822-2-CHEM

UF822-5-CHEM

UF822-5-CHEMUF822-6-CHEM

UF822-6-CHEM

UF823-1-CHEM

UF823-1-CHEMUF823-1-CHEMUF823-2-CHEMUF823-2-CHEM

UF823-5-CHEM

UF823-5-CHEM

UF823-6-CHEMUF823-6-CHEM

UF824-1-CHEM

UF824-1-CHEM

UF824-2-CHEMUF824-2-CHEM

UF824-5-CHEM

UF824-5-CHEM

UF824-6-CHEMUF824-6-CHEM

UF825-1-CHEM

UF825-1-CHEM

UF825-2-CHEMUF825-2-CHEM

UF825-5-CHEM

UF825-5-CHEMUF825-6-CHEMUF825-6-CHEM

UF826-1-CHEM

UF826-1-CHEM

UF826-2-CHEMUF826-2-CHEM

UF826-6-CHEM

UF826-6-CHEM

UF827-1-CHEM

UF827-1-CHEM

UF827-2-CHEMUF827-2-CHEM

UF827-5-CHEM

UF827-5-CHEM

UF827-6-CHEMUF827-6-CHEM

UF828-1-CHEM

UF828-1-CHEMUF828-2-CHEM

UF828-2-CHEM

UF829-1-CHEM

UF829-1-CHEM

UF829-2-CHEMUF829-2-CHEM

UF829-5-CHEM

UF829-5-CHEM

UF829-6-CHEMUF829-6-CHEM

UF830-5-CHEM

UF830-5-CHEM

UF830-6-CHEMUF830-6-CHEM

UF831-1-CHEM

UF831-1-CHEM

UF831-2-CHEM

UF831-2-CHEM

UF831-5-CHEM

UF831-5-CHEM

UF831-6-CHEM

UF831-6-CHEM

UF832-1-CHEM

UF832-1-CHEM

UF832-2-CHEM

UF832-2-CHEM

UF832-5-CHEM

UF832-5-CHEM

UF832-6-CHEMUF832-6-CHEM

UF833-1-CHEM

UF833-1-CHEM

UF833-2-CHEMUF833-2-CHEM

UF833-5-CHEMUF833-5-CHEM

UF833-6-CHEMUF833-6-CHEM

UF834-2-CHEMUF834-2-CHEM

UF834-5-CHEM

UF834-5-CHEMUF834-6-CHEM

UF834-6-CHEM

UF835-1-CHEM

UF835-1-CHEM

UF835-2-CHEMUF835-2-CHEM

UF835-5-CHEM

UF835-5-CHEM

UF835-6-CHEMUF835-6-CHEM

UF836-1-CHEM

UF836-1-CHEMUF836-1-CHEM

UF836-2-CHEM

UF836-2-CHEM

UF836-5-CHEM

UF836-5-CHEM

UF836-6-CHEM

UF836-6-CHEM

UF837-1-CHEMUF837-1-CHEM

UF837-2-CHEM

UF837-2-CHEM

UF837-5-CHEM

UF837-5-CHEM

UF837-6-CHEMUF837-6-CHEM

UF838-1-CHEM

UF838-1-CHEM

UF838-2-CHEMUF838-2-CHEM

UF838-5-CHEM

UF838-5-CHEM

UF838-6-CHEMUF838-6-CHEM

UF839-1-CHEM

UF839-1-CHEM

UF839-2-CHEMUF839-2-CHEM

UF839-5-CHEM

UF839-5-CHEMUF839-6-CHEM

UF839-6-CHEMUF840-1-CHEM

UF840-1-CHEM

UF840-2-CHEM

UF840-2-CHEMUF840-5-CHEM

UF840-5-CHEM

UF840-6-CHEMUF840-6-CHEM

UF841-1-CHEM

UF841-1-CHEM

UF841-2-CHEMUF841-2-CHEM

UF841-5-CHEM

UF841-5-CHEM

UF841-6-CHEM

UF841-6-CHEM

UF842-1-CHEM

UF842-1-CHEMUF842-1-CHEMUF842-2-CHEM

UF842-5-CHEM

UF842-5-CHEM

UF842-6-CHEM

UF842-6-CHEM

UF844-1-CHEM

UF844-1-CHEMUF844-2-CHEMUF844-2-CHEM

UF844-5-CHEM

UF844-5-CHEMUF844-6-CHEM

UF844-6-CHEM

UF845-1-CHEM

UF845-1-CHEM

UF845-2-CHEM

UF845-2-CHEM

UF845-5-CHEM

UF845-5-CHEMUF845-6-CHEM

UF845-6-CHEM

UF846-1-CHEMUF846-1-CHEM

UF846-1-CHEMUF846-2-CHEM

UF846-2-CHEM

UF846-5-CHEM

UF846-5-CHEM

UF846-6-CHEM

UF846-6-CHEM

UF847-1-CHEM

UF847-1-CHEM

UF847-2-CHEM

UF847-2-CHEM

UF847-5-CHEM

UF847-5-CHEM

UF848-1-CHEM

UF848-1-CHEM

UF848-2-CHEMUF848-2-CHEM

UF848-5-CHEM

UF848-5-CHEMUF848-6-CHEMUF848-6-CHEM

UF849-1-CHEM

UF849-1-CHEM

UF849-2-CHEMUF849-2-CHEM

UF849-5-CHEM

UF849-5-CHEM

UF849-6-CHEMUF849-6-CHEM

UF850-1-CHEM

UF850-1-CHEM

UF850-2-CHEM

UF850-2-CHEM

UF850-5-CHEM

UF850-5-CHEM

UF850-6-CHEM

UF850-6-CHEM

UF852-1-CHEM

UF852-1-CHEM

UF852-2-CHEMUF852-2-CHEM

UF852-5-CHEM

UF852-5-CHEM

UF852-6-CHEM

UF852-6-CHEM

UF853-1-CHEM

UF853-1-CHEM

UF853-2-CHEMUF853-2-CHEM

UF853-5-CHEM

UF853-5-CHEM

UF853-6-CHEMUF853-6-CHEM

UF854-1-CHEM

UF854-1-CHEM

UF854-2-CHEMUF854-2-CHEM

UF854-5-CHEM

UF854-5-CHEM

UF854-6-CHEM

UF854-6-CHEM

UF855-1-CHEMUF855-1-CHEM

UF855-2-CHEM

UF855-2-CHEMUF855-5-CHEM

UF855-5-CHEM

UF855-6-CHEMUF855-6-CHEM

UF856-1-CHEM

UF856-1-CHEM

UF856-2-CHEM

UF856-2-CHEM

UF856-5-CHEM

UF856-5-CHEM

UF856-6-CHEMUF856-6-CHEM UF857-1-CHEM

UF857-1-CHEM

UF857-2-CHEMUF857-2-CHEM

UF857-5-CHEM

UF857-5-CHEM

UF857-6-CHEMUF857-6-CHEM

UF858-1-CHEM

UF858-1-CHEMUF858-2-CHEMUF858-2-CHEM

UF858-5-CHEM

UF858-5-CHEMUF858-6-CHEMUF858-6-CHEM

UF859-1-CHEM

UF859-1-CHEM

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PC1

PC2 Scores

Sykes et al, Novaes et al

• HTP compositional analysis– Glucan and xylan content

– Correlates with wet chemistry

• 24 samples in 4 hours- single reactor plate– 2-3 days by wet chemistry

– Expandable to multiple plates

• Native or pretreated biomass

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

0.00 20.00 40.00 60.00 80.00

% C

arbo

hydr

ate-

HTP

% Carbohydrate-Wet Chemistry

prtx glucan prtx xylanstd glucan std xylan

Poplar Association Study-1st 2 Sets

• Error bars represent standard deviation of triplicate runs – reps equally distributed throughout plate; 24 samples per plate; – red bars are samples repeated in multiple plates

• All plate normalized to BESC poplar standard– 8 standard runs per plate

• Data didn’t look very encouraging at first……

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

16.0 18.0 20.0 22.0 24.0 26.0 28.0

suga

r rel

ease

-g/g

bio

mas

s

% Lignin

Total Sugar

Glucose

Xylose

Poplar Association Study-755

•No apparent impact of lignin content on sugar release after pretreatment/enzyme hydrolysis

Selig et al

0.0

0.1

0.1

0.2

0.2

0.3

0.3

0.4

0.4

0.5

0.5

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5

suga

r rel

ease

-g/g

bio

mas

s

S/G Lignins

Glucose

Xylose

Poplar Association Study- 755

•Glucan and xylan conversions show trend with S/G

•Xylan conversion is somewhat less scattered

Selig et al

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…B

ES

…

Com

bine

d Su

gar R

elea

se (g

/g b

iom

ass)

run #1run #2

Poplar 4-CL Study- 100

• System consistently identifies outliers within small & large sample sets

Voelker et al

Danish Wheatstraw- 1130

• 1100 sample set – 25 separate experimental sets; strains, harvest conditions…etc• Collaboration with Claus Felby, U. Copenhagen

• Consistent overlap between plates • Variation between sets and not plates

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0 240 480 720 960

Co

mb

ine

d S

uga

r R

ele

ase

(g/

g b

iom

ass)

Noble Foundation Switchgrass-58

• 58 samples included controls and replicates

• SSF carried out at ORNL• Jonathon Mielenz lab

• First demonstration of fermentation/HTP pipeline correlation

0.000

0.100

0.200

0.300

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0.500

0.600

mg/

mL

suga

rSugar Release - Pretreatment + Saccharification

glucose xylose

200

220

240

260

280

300

320

340

360

0.150 0.200 0.250 0.300 0.350 0.400

mg

etha

nol/g

dry

bio

mas

s

recalcitrance screening glucose release in mg/g dry biomass

SSF vs. Recalcitrance Screening

Summary

• First high-throughput thermochemical pretreatment/enzyme digestion pipeline– We have run dozens of campaigns – ~10,000 samples analyzed

• Solved numerous obstacles– Biomass processing and dispensing– Extractives and starch variability– Pressure and evaporation control– Rapid sugar analysis – HTP chemical analysis

• Severe pretreatment conditions possible– High temperatures (150-200oC), pressures, low pH

• HTP Analytical chemistry for lignin/carbohydrate content– MBMS for total lignin, S/G ratios– HTP compositional analysis for cellulose and xylan content– Starch analysis is possible

• Data is comparative within sample sets– Determine correlations of recalcitrance with environmental factors– Measure effects of genetic variation– Find outliers in large sample populations

• Capacity of 100s of samples/day– Current bottleneck is sample preparation

Acknowledgments

• Stephen Decker• Melvin Tucker• Mike Himmel• Rob Sykes• Kristen Gracom• Crissa Doeppke• Melissa Carlile• Cody Law• Geoffrey Turner• Mark Davis

• Charley Wyman• Michael Studer• Jaclyn DeMartini• Heather McKenzie

This work was supported by the U.S. Department of Energy under Contract No. DE-AC36-08-GO28308 with the National Renewable Energy Laboratory.The BioEnergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

• Martin Keller• Brian Davison• Gerry Tuskan• Jonathon Mielenz

• Rick Nelson• Fang Chen

• William Everitt• http://aspenmachining.com

• Claus Felby

References• Somerville, C., S. Bauer, G. Brininstool, M. Facette, T. Hamann, J. Milne, E. Osborne, A. Paredez, S.

Persson, T. Raab, S. Vorwerk, H. Youngs, Toward a systems approach to understanding plant cell walls.Science 306, 2206-2211 (2004).

• Sykes, R., Yung, M., Novaes, E., Kirst M., Peter,G., Davis, M., “High Throughput Screening of Plant CellWall Composition Using Pyrolysis Molecular Beam Mass Spectroscopy” in Biofuels: Methods andProtocols, Methods in Molecular Biology, 581, Mielenz, J.R.,(ed ) Humana Press pp 169-183 (2009).

• Novaes,E., Osorio,L., Drost, D.R., Miles, B.L., Boaventura-Novaes C.R.D.,Benedict,C., Dervinis, C., Yu, Q., Sykes R., Davis, M., Martin, T.A., Peter, G.F., Kirst, M., “Quantitative genetic analysis of biomass and wood chemistry of Populus under different nitrogen levels”, New Phytologist , 182 (4), 878-890 (2009).

• Voelker, S. L., B. Lachenbruch, F. C. Meinzer, M. Jourdes, C. Ki, A. M. Patten, L. B. Davin, N. G. Lewis, G. A. Tuskan, L. Gunter, S. R. Decker, M. J. Selig, R. Sykes, M. E. Himmel, P. Kitin, O. Shevchenko, and S. H. Strauss. 2010 (in press). “Antisense down-regulation of 4CL expression alters lignification, tree growth and saccharification potential of field-grown poplar”. Plant Physiol.

• Selig, M. J., M. P. Tucker, R. W. Sykes, K. L. Reichel, R. Brunecky, M. E. Himmel, M. F. Davis, and S. R. Decker. (2010). “Biomass Recalcitrance Screening by Integrated High Throughput Hydrothermal Pretreatment and Enzymatic Saccharification”. Ind. Biotechnol. 6:104-111

• Decker, S. R., R. Brunecky, M. P. Tucker, M. E. Himmel, and M. J. Selig. (2009). “High Throughput Screening Techniques for Biomass Conversion”. Bioenergy Research. 2:79-192.