Enzymatic transformations Wheat bran - ValBiom · Pr. Sandrine BOUQUILLON, Université de Reims...

Pr. Sandrine BOUQUILLON, Université de Reims6èmes Rencontres de la Biomasse

Gembloux – 21 octobre 2009

1

PentoraffineriePentoraffinerie et chimie des pentoseset chimie des pentoses

Sandrine BOUQUILLONSandrine BOUQUILLON

Wheat bran

Enzymatic transformations

Pentoses :D-xylose

L-arabinose

Arabino-xylo-oligosaccharides

Pentoses syrup



2

Enzymatic hydrolysis of hemicelluloses from wheat bran

Wheat bran

Xylanase (Thermobacillus xylanilyticus)

Arabino-xylo-oligosaccharidesX2, X3, X4A, > DP

Xylosidases Arabinosidase (Thermobacillus xylanilyticus)

Structural resolution of the arabinosidasefrom T. xylanilyticus

Arabinose + xylose

Production and characterization of new xylosidases from Bacillus halodurans

Improving thermostability of xylanase

from T. xylanilyticus

2008. Biochemistry 2008, 47, 7441.

Appl. Microbiol. Biotechnol. 2006, 73, 582.

Bioresource Biotechnology 2009, 100, 338.

UMR FARE INRAC. RémondM. O’Donohue

Enzymatic hydrolysis of hemicelluloses from wheat bran

Hydrolysis of wheat bran with an xylanase at laboratory scale (g)

(UMR FARE)

Hydrolysis of wheat bran with an xylanase at laboratory scale (g)

(UMR FARE)

Transposition at semi-pilot and pilot scales

(ARD)

Transposition at semi-pilot and pilot scales

(ARD)

Xylanase Tx-Xyl40 % of xylans are hydrolysed into XOs

Enzymatic hydrolysis versus acid hydrolysis: high purity of pentoses syrup, no furfural molecules produced

Yields of enzymatic hydrolysis of xylans are identical (40 %) at the different scales

ARDF. Martel




3

Pentoses

D-xyloseL-arabinose

Pentosides

Surfactants

Bran sirup

Enzymatic and chemical transformations

Enzymatic synthesis of alkyl arabinosides

pNP α-L-Araf

or X4A Alcools

Arabinosidasede T. Xylanilyticus

Methyl α-L-arabinoside, 55 %Ethyl α-L-arabinoside, 54 %

isopropyl α-L-arabinoside, 24 %n-butyl α-L-arabinoside, 42 %2-butyl α-L-arabinoside, 41 %

Allyl α-L-arabinoside, 33 %

OHO

OHHO

On

+

Tetrahedron Lett. 2002, 43, 9653.

Carbohydr. Res., 2004, 339, 2019.

Alkyl arabinosides

First arabinosidase described for the enzymatic synthesis of arabinose-based molecules

ICMR UMR CNRSR. Plantier RoyonC. PortellaC. Bliard




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Enzymatic synthesis of alkyl arabinosidesUMR FARE INRAC. RémondM. O’Donohue

ICMR UMR CNRSR. Plantier RoyonM. Muzard

Alkyl xylosides

pNP β-D-Xylp Alcools

Xylosidase GH39 de Bacillus halodurans

+ OH

OHO

HO On

Methyl β-D-xyloside, 10,7 %Ethyl β-D-xyloside, 9 %

n-propyl β-D-xyloside, 23,4 %n-butyl β-D-xyloside, 14,9 %n-pentyl β-D-xyloside, 2,5 %

Wheat bran

20 g

(8 g xylans) Xylanase

Tx-Xyl11

OH

OHO

HO

OH

OHO

OOH

OH

OHO

HO

OH

OHO

O

OH

OHO

OOH

+ arabino-xylooligosaccharides

3,15 g

Xylosidase

XylBH39

OHO

HO

OH

O

Propyl β-D-xylopyranoside

965 mg

n- propanol 5%

A two-steps synthesis of propyl β-D-xyloside from wheat bran

J. Mol. Catal. B: Enzymatic, 2009, 58, 1.

ARD WheatoleoB. Estrine

Wheat Bran and Straw

- Plant oils : coco, pko , palm ,- Sugar beet, wheat…

Xylose ArabinoseGlucose

Hydrophilic head Lipophilic tail

Glycosidic bond

Synthesis of surfactants



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ARD WheatoleoB. Estrine A complete range of A complete range of ““greengreen”” products in order to meet all products in order to meet all

formulatorformulator’’s needs (hydrotropes, surfactants and emulsifiers)s needs (hydrotropes, surfactants and emulsifiers)

Short chain products (C4-C8)

Solubilizer / HydrotropeR/6505, …..

Wetting / Foaming Degreasing SurfactantsR/6781, …

EmulsifiersR/6669, …

Middle chain products (C8-C14)

Long chain products (C14-C22)


ARD WheatoleoB. Estrine RadiaRadia®®EasysurfEasysurf : : Properties and applications

Glucose Glucose basedbased SS. . Pentose basedPentose based SS..

HO O

HOOH

OCnH2n+1

OH

m

R1O

HOOH

O (CH2)n CH3

R2

m'

Glucosides(wheat - starch origin)

Biodegradable

Mildness

Pentosides(Wheat bran and straw origin)

Biodegradable

Mildness

HLB

CMC

Solubility

Degreasing

γ O/W

Wetting

Can’t be solubilized in oils Oil solubility




6

RadiaRadia®®EasysurfEasysurf: building : building pentosespentoses based based surfactant according a surfactant according a biorefinerybiorefinery conceptconcept

BiorefineryBiorefinery conceptconcept : : valorization of non food agricultural byvalorization of non food agricultural by--products rich in products rich in pentosespentoses..

Alkyl Poly Alkyl Poly PentosidesPentosides : : new generation of sugar based surfactantsnew generation of sugar based surfactants

-- low environmental impactlow environmental impact-- rapid and complete biodegradabilityrapid and complete biodegradability-- low low ecotoxicecotoxic behavior behavior -- powerful performance capabilitiespowerful performance capabilities-- ideal candidates for a broad range of applicationsideal candidates for a broad range of applications

ARD WheatoleoB. Estrine RadiaRadia®®EasysurfEasysurf : : Properties and applications


2 processes (homogeneous catalysis)

Telomerization of butadiene with pentoses

French Patent CNRS 2001, N° 0116363, 2003, PCT WO 03053987.Eur. J. Org. Chem. 2004, 2914.Green Chem. 2005, 7, 219.J. Mol. Cat. 2005, 244, 93.

Sugar / Pd = 150PPh3 / Pd = 3Butadiene/Pd = 900DMF, 75 °C, 1 hHigh [pentose]

up to 95% conversion of sugar≈ 70 % monooctadienyl-xylosides

Sugar / Pd = 150Amine / Pd = 150TPPTS / Pd = 3H2O, 80°C, 1 hHigh [pentose]

up to 99% conversion of sugar≈ 70 % monooctadienyl-xylosidesOr ≈ 60 % dioctadienyl-xylosides

OOH

HO OH

OO

HO

HO

OH+ isomers

HOPd(OAc)2 / PPh3

DMF

OOH

HO OHO

O

HO O

H2O

HO

HO+ isomers

Pd(acac)2TPPTS

C12H25NMe2

EtNPr2i

D-xyloseL-arabinose


ICMR UMR CNRSS. BouquillonJ. Muzart



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Surfactant behavior - monooctadienyles : “medium”- dioctadienyles : “good”- crude mixture : “good”

Crude mixture :6% Xylose, 62% Monooctadienyles, 27% Dioctadienyles, 5% Trioctadienyles

Carbohydr. Res. 2006, 341, 1938.

Useless separations

Application to bran syrup

(bran syrup = xylose + arabinose + glucose + …)

Composition of the mixture1% Bran syrup67% Monooctadienyles31% Dioctadienyles1% Trioctadienyles

20,00

25,00

30,00

35,00

40,00

45,00

50,00

55,00

60,00

65,00

70,00

1,00 10,00 100,00 1000,00

concentration (mg/L)

surf

ace

tens

ion

(mN

/m)

“Good” behavior as surfactant



Glycosylations

Optimal conditions (n = 0, 5, 6) : PTSA, alcohol, 20 h, 70% ≤ yields ≤ 80%

Simple and rapid syntheses of surfactants- monocatenar- bolaform

From free D-xylose

Metatheses

C10, C18, C20

Grubbs I [Ru] 7% mol 3 h, 40°Cn

n = 1, 5, 6O

O

OH

HO

HOO

OOO

OH

HO

HO

HO

OH

OH

n nα or β

αα or ββ 57-90%

OOH

HO OH

HO + HO "H+"O

O

HO OH

HOn n

n = 1, 5, 645ºC-80ºC, 24 h

"H+" = Amberlite IR 120 resin, PTSA, AcCl

Yields = 18-81%

Carbohydr. Res. 2007, 342(2), 154.





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Pentoses

D-xyloseL-arabinose

« Fine chemicals »

OHO

OHHO OH

Chemical transformations

Preparation of useful building-blocks

Objective: to develop transformations leading to common intermediates from either of the sugars

D-xylose

L-arabinose

CHOOH

HOOH

CH2OH

CHOOH

HOHO

CH2OH

4

4

OP2P3O

O

O

OP2OP3

OP1X

O

OP2OP3

OP1X

An interestingfunctionnalized intermediate

Reductiveelimination

- Nucleophilic addition

- Olefination

- Intramolecular reactions

- Reductive amination

suppression of the chiral character of C-4

ICMR UMR CNRSC. PortellaR. Plantier Royon



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« Domino » Reactions with organometallic species

O

BnO

I

O

O

OHBnO

HOR

solvent, 0°C

RLi ou RMgBr

~90%

R = Me, n-Bu, iPr, Allyl, Ph

dr = 4/1 to 7/1

OHBnO

O

BnO

I

O

O Y Z

Me3Si LiOHBnO

+OHBnO

OHBnO

OHBnO

Y

Z

S

S

SMe

SMe

SPh

52 - 86%

Large variety of building blocksSimilar results from xylose or arabinose derivatives

J. Org. Chem. 2007, 72, 2271.Carbohydr.Res. 2007, 342, 2450.

ICMR UMR CNRSC. PortellaR. Plantier Royon

Synthesis of heterocyclic compoundsICMR UMR CNRSC. PortellaR. Plantier Royon

O

ORRO ORRO

YYCH2P(O)(OEt)2Et3N / LiCl

N

R

Y

OBn

OBn

*

Fonctionnalization

,Objective: New glycosidase inhibitors

D-xyloseand/or

L-arabinose THF



10

Tetrahedron lett. 2009, 50(17), 1902.

Extension to 2nd and 3rd generations and to L-arabinose

O

OHHOOH

O

OAcAcO O

OH

HO AcO2 steps (60 %)

S

Cs2CO3 (4 eq. / Cl atom)O CH

N NMe

PS

ClClN3P3

6

O

AcO OAcO

OAc

O CHN N

Me

PS O

N3P3O

AcO OAc

OOOAc

6

THF, 24 h, 0°C to RT

S (2 eq. / Cl atom)

MeONa (0.5 M)MeOH/CH2Cl2, RT, 12h

O

AcO OHO

OH

O CHN N

Me

PS O

N3P3O

HO OH

OOOH

6

1st generationQuantitative

Synthesis of Phosphorous-based GlycodendrimersICMR UMR CNRSS. Bouquillon

PN P N

PN OO

O O

OO

N

N

NN

N

N

N

NN

N

N

N P

P

PP

P

PS

OO

O

O

S O

OS

O S

SO

S

O

O

O

O

N

N

NN

NN

N

N

N

NN

NN

N

N

N

N

NN

N

N

N

N

N

P

P

P

PP

P

P

P

P

P P

P

SO

O

O

O

S

S OO

SO

O

O

S

O

O

S

O

O S

OO

S

O O

S

O O

S

O

OO

S

S O

O O

O

O

O

OO

O

O

OO

O

O

O

O

OO

OO

OR

ORRO

OR

OR

O

ROOR

OR

RO OR

OR

ROOR

OR

O

O

O

O

O

O

O

O

O

O

OO

OO

O

O

O

O

O

OO

O

O

O

O

O

O

O

OR

OR

OR

OROR

ORRO

ROOR

OR

OR

OR

RO

ROOR

OR

OROR

RO

RO

RO

ORRO

RO

ORRO

RO

ORRO

RO

ORRO

RO

ORRO

RO

RO

RO

RO

ORRO

RORO

RO

RO

ORRO

RO

RO

ROOR

OR

OR

OR

O O

ROOR

OR

Generation 2

Applications :CatalysisNanoencapsulation

R = H, OAc

Synthesis of Phosphorous-based GlycodendrimersICMR UMR CNRSS. Bouquillon



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J. Org. Chem. 2009, in press.

Extension to 2nd and 3rd generations

Si O

OO

O

Si

Si

O

O

O

O

OO

O

O

OO

Ph

3

NNN

O S"

NNN

OS"

N NN

OS"

NN

NO

S"

NN

NO S"

NN

NO S"

N NN

O S"

N NN

OS"

NN

NO

S"

S' (2 eq. / branch)

O

OAcO

AcO

OAc

N3

O

HO

HO

OH

Cu(SO4)2 . 5H2O ( 4 eq. / branch)

Sodium ascorbate (8 eq. / branch)Si O

OO

OO

Si

Si

O

O

O

O

OO

O

O

OO

Ph

3

OH

5 steps (40 %)

S'

S" =O

AcO

AcO

OAc

THF / water, RT, 12 h

59%

"Click" Chemistry

Synthesis of Silicium-based GlycodendrimersICMR UMR CNRSS. Bouquillon

J. Org. Chem. 2009, 74, 5071

Applications :CatalysisNanoencapsulation

N NN

OO

O

N NN

N NN

O

O

Si

N NN

OO

O

N NNN N

N

O

O

Si

N NN OO

O

N NNN N

N

O

O

Si

OSi

N NN O

O

O

N NN

N NN

O

O

Si

N NN

OO

O

N NN

N NN

O

OSi

NN

N

O

O

O

NN N

NN

N

OO Si

O Si

NN N

O

O

O

NN N

NN N

O O Si

NN N

O

OO

NN N

NN N

O

OSi

NN N

O

OO

NN N

NN N

O

OSi

OSi

NN N

O

O ONN N

NN N

O

O

Si

NN

N

O

O ON

NN

NN

N

O

O

Si

NN

N

O

O O

NN

N

NN

N

O

O

Si

O

Si

NN

N

O

OO

NN

N

NN

N

O

O

Si

NNN

OO

O

NNN

NNN

O

O

Si

NNN

OO

O

NNN

NNN

O

O

Si

O

Si

NNN

O O

O

NNN NN

N

O

O

Si

NNNO O

O

NNN

NNN

O

O

Si

NNN

OO

O

NNN

NNN

O

OSi

OSi

NN

N

O

O

O

NN

N

NN

N

OOSi

NNN

O

O

O

NNN

NNN

OOSi

NNN

O

O

O

NNN

NNNO

OSi

OSi

NNN

O

O

O

NNN

NNNO

OSi

NNN

O

OO

NNN

NNN

O

OSi

NNN

O

OO

NNN

NNN

O

O

Si

O

Si

NN

N

O

OONN

N

NN

N

O

O

Si

NN

N

O

OO

NN

N

NN

N

O

O

Si

NN

N

OO

O

NN

N

NN

N

O

O

Si

O

Si

RO

RO

RO

RO

RORO

RORO

RO

RORO

RORO RO RO RO RO RO

OROR

OROR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OR

OROR

OROR

OROROROR

RORO

RORO

RORO RO RO RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

RO

O

OAcO

AcO

OAc

RO

Generation 3

Synthesis of Silicium-based GlycodendrimersICMR UMR CNRSS. Bouquillon



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Conclusions

Wheat bran

Pentoses :D-xylose

L-arabinose

Arabino-xylo-oligosaccharides

Pentoses syrup « Fine chemicals »

Pentosides

Surfactants

ICMR UMR CNRSARD WheatoleoUMR FARE INRA

Enzymatic transformations Wheat bran - ValBiom · Pr. Sandrine BOUQUILLON, Université de Reims...

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Transcript of Enzymatic transformations Wheat bran - ValBiom · Pr. Sandrine BOUQUILLON, Université de Reims...