An Auspicious Application of Laccase and Hydrogen ... ISWPCf.pdf · Recalcitrant Paper Dyes ......
Transcript of An Auspicious Application of Laccase and Hydrogen ... ISWPCf.pdf · Recalcitrant Paper Dyes ......
An Auspicious Application An Auspicious Application of Laccase and of Laccase and
Hydrogen Hydrogen PeroxidasesPeroxidasesfor Biobleaching of for Biobleaching of
Recalcitrant Paper DyesRecalcitrant Paper Dyes
Kristina Knutson and Arthur Ragauskas, Kristina Knutson and Arthur Ragauskas, Institute of Paper Science and Institute of Paper Science and
Technology, Atlanta, GATechnology, Atlanta, GA
•81 million tons of waste fiber in US in 1994
>90% linerboard
>80% newsprint
>60% white office ledger
<30% mixed office waste
Recycling Continues to Increase
0
20
40
60
1987 1994 1998 2003 2012
% R
ecyc
led
Research ObjectivesResearch Objectives
Test effectiveness of virgin pulp Test effectiveness of virgin pulp biobleaching technologies on color biobleaching technologies on color removal from dyed pulpsremoval from dyed pulps
+ Enzyme
TodayToday’’s Talks Talk
Dyes Direct Yellow 11 & Basazol 46LDyes Direct Yellow 11 & Basazol 46LMeasurement of ColorMeasurement of ColorLaccase & Laccase & peroxidasesperoxidasesPeroxidasesPeroxidases experimental resultsexperimental resultsLaccase experimental resultsLaccase experimental resultsSummarySummary
Recycling Colored PapersRecycling Colored Papers
20.220.222.822.840.540.515.015.0GoldenrodGoldenrod71.071.061.561.572.372.354.354.3File StockFile Stock
79.779.769.069.080.780.765.565.5Colored Colored LedgerLedger
81.881.866.766.781.581.562.162.1Mixed Mixed Office Office WasteWaste
Hydrosulfite Hydrosulfite (Dithionite) (Dithionite)
2%2%
Hydrogen Hydrogen Peroxide Peroxide
1%1%
FormamidineFormamidinesulfinic acid sulfinic acid (FAS) 0.5% (FAS) 0.5%
Original Original DeinkedDeinked
Raw Raw Material Material TypeType
Brightness After TreatmentBrightness After Treatment
Reference: Ferguson, L.D. 1996 TAPPI Deinking Short Course “Bleaching Wastepaper”
Structure Direct Yellow 11Structure Direct Yellow 11
Structure most problematical dye commonly used for office paper
Structure Basic Dye Structure Basic Dye BaszolBaszol 46L46LUnited States Patent Number 4,256,458 March 17, 1981
Methine Dyes for Paper and Amonically-Modified FibersPatent applies to methine dyes of the formula
•A is an organic or inorganic ion•R is hydrogen, methyl or ethyl•Q groups are the same or different substituted or unsubstituted aryl or hetaryl radicals•X represents a bridge member selected from the group consisting of aliphatic,
aromatic and heteroaromatic radicals.
2A-
Structure of most common yellow dye used in directory paper
Example SpectraExample Spectra
ph8.5 No PEG+B40 Con
-0.2
0
0.2
0.4
0.6
0.8
1
200 300 400 500 600
Wavelength
Abs
orba
nce
Calculation of Dye Removal IndexCalculation of Dye Removal IndexDistance from Ideal Bleach Point = R2
R2 = a2 + b2 + (100-L) 2
Amount of Color Removal = -∆R2
∆R2 = R22 – R1
2
∆R2 = (a22 – a1
2) + (b22 – b1
2) + [(100-L2) 2 – (100-L1) 2]
Dye Removal Index (DRI) = -100 [∆R2 / R1
2] DRI = Percent Color Removed
by the Treatment
L=100 a=b=0
R2
R1
a = b = L = 0
Potential of Biobleaching Potential of Biobleaching Recycled FiberRecycled Fiber
N OHC
O
H3C
MEDIATORMEDIATOR
Oxidation of Oxidation of ßß--OO--4 lignin 4 lignin dimerdimer I I by laccase/HBT systemby laccase/HBT system
Peroxidase Catalytic CyclePeroxidase Catalytic Cycle
Clear applications for color destruction that have not yet been examined
Horseradish Peroxidase SystemHorseradish Peroxidase System
2 hour Reaction at 452 hour Reaction at 45°°C C 100mM Acetate buffer, 100mM Acetate buffer, pH 3.8pH 3.811--mM Veratryl alcohol mM Veratryl alcohol 10ppm Dye10ppm Dye5 Units Horseradish 5 Units Horseradish peroxidase /ml peroxidase /ml 0.1 mM Hydrogen 0.1 mM Hydrogen peroxideperoxide
Peroxidase Decolorizes Basic DyePeroxidase Decolorizes Basic Dye
HRP Treatment
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
280 330 380 430 480 530 580
Wavelength (nm)
Abs
orba
nce
Basazol 46L
Treated Basl 46L
Direct Yellow 11
Treated Yellow11
Soybean Peroxidase SystemSoybean Peroxidase System
2 hour Reaction at 452 hour Reaction at 45°°C C 100mM Acetate buffer, pH 3.8100mM Acetate buffer, pH 3.811--mM Veratryl alcohol mM Veratryl alcohol 0.1M Calcium chloride0.1M Calcium chloride10ppm Dye10ppm Dye1 Units Soybean peroxidase 1 Units Soybean peroxidase /ml /ml 0.1 mM Hydrogen peroxide0.1 mM Hydrogen peroxide
Comparison of SBP and HRPComparison of SBP and HRP
-20
0
20
40
60
80
100
Direct Yellow11
Basazol 46L
% R
educ
tion
in A
bs A
rea
SBP HRP
SBP Effective Over Wide Ph RangeSBP Effective Over Wide Ph Range
89.489.489.789.789.089.08.58.5
90.690.687.487.489.789.74.54.5
86.986.985.385.385.185.13.83.8
67.367.335K35K
60.260.210K10K
69.269.2NonNon
ee2.42.4Basazol Basazol
46L46L
% % DecDec
..PEGPEG% Dec.% Dec.PEGPEG
% % DecDec
..PEGPEGpHpHSBP TreatSBP Treat
LaccaseLaccase--Mediator SystemMediator SystemFor Treating Dyed PulpFor Treating Dyed Pulp
2 hour Reaction at 452 hour Reaction at 45°°C C 10bar Oxygen pressure10bar Oxygen pressure100mM Acetate buffer, 100mM Acetate buffer, pH 4.5pH 4.5Mediator 0.005Mediator 0.005--5mM 5mM 50 50 µµkatkat/mL Laccase/mL Laccase10ppm Direct Yellow 11 10ppm Direct Yellow 11 oror3% pulp consistency3% pulp consistency
Laccase MediatorsLaccase Mediators
ABTSABTS2,2'azinobis2,2'azinobis--33--ethylethyl--benzthiazolinebenzthiazoline--55--sulfonatesulfonate
Violuric acidVioluric acid
HBTHBT11--Hydroxy Hydroxy benzotriazolebenzotriazole
Mediator StructureMediator StructureMediator NameMediator Name
S
N
S
N
H2C CH3 CH2H3C
N N
SO3HHO3S
HN NH
O
OO
NOH
N
N
N
OH
Dissolved Direct Yellow 11 + LMSDissolved Direct Yellow 11 + LMS
0
0.5
1
1.5
2
2.5
3
200 300 400 500 600
Wavelength, nm
20 ppm Direct Yellow 11
Abs
orba
nce
Dye, 1 mM HBT, Lac, to
Dye, HBT, Lac, 2hr 45°C, 10bar O2
Effectiveness of LMS Decolorization Effectiveness of LMS Decolorization Depends on MediatorDepends on Mediator
Mediator Comparison
0
10
20
30
40
50
60
70
Lac+HBT Lac+VA Lac+ABTS
DR
I (D
ye R
emov
al In
dex)
ABTS Effective at Low ABTS Effective at Low ConcentrationConcentration
Dose Dependence
010203040506070
0 0.005 0.01 0.05 0.5 5
mM ABTS
DR
I (D
ye R
emov
al In
dex)
PostPost--LMS Reaction Liquid PhaseLMS Reaction Liquid Phase
Effluents from Treated Pulps
0
0.2
0.4
0.6
0.8
1
240 280 320 360 400 440 480
Wavelength (nm)
Abs
orba
nce
White Pulp +Lac+0.1mM ABTSDyed Pulp +Lac+0.1mM ABTS
Laccase + ABTS
Direct Yellow 11 + Laccase + ABTSAfter 2hr at 10bar O2, 45 ºC
Purified Direct Yellow 11
ConclusionConclusion
Laccase/ABTS system can decolorize stilbene Laccase/ABTS system can decolorize stilbene dye Direct Yellow 11dye Direct Yellow 11PeroxidasesPeroxidases are ineffective for Direct Yellow 11are ineffective for Direct Yellow 11PeroxidasesPeroxidases do decolorize basic dye Basazol 46Ldo decolorize basic dye Basazol 46LLaccase decolorizes Basazol 46LLaccase decolorizes Basazol 46L