Synthetic analogues of biopolyesters - present and future

Marek M. Kowalczuk

MONOMERS

OLIGOMERS

POLYMERS

3-HB (3-hydroxy - butyric acids)

Amino acids

Monosacharides

Nucleotides

cP(3-HB) (channel component)

Oligopeptides Oligosacharides Oligonucleotides

sP(3-HB) (cf. storage)

Polypeptides (enzymes, silk)

Polysacharides (cf. cellulose, starch)

Polynucleotides (cf. DNA, RNA)

BIOPOLYMERS

Bacterial synthesis of Poly(3-hydroxyalkanoate)

Short-chain-length PHA Medium-chain-length PHA

Thermoplasts (Tm: 120 – 180°C)

•High crystallinity (40-80%)

•Density >1.2 g cm-3

•Elongation at break 6-10%

•Industrial production: +

•Elastomeric, thermoplastic, duroplastics or

fluidoplastic properties (Tm: n.d. – 80°C)

•Low crystallinity (0-40%)

•Density: ca. 1.05 g cm-3

•Elongation at break: Adjustable (>100%)

High Levels of Bioplastic Are Produced in Fertile Transplastomic Tobacco Plants Engineered with a Synthetic Operon for the Production of Polyhydroxybutyrate

Plant Physiology, April 2011, Vol. 155, pp. 1690–1708

Plastid-encoded expression of the PHB multigene pathway can produce high levels of PHB in tobacco without affecting its fertility and that the engineered pathway is stably transferred to the next generation.

Downstream processing of mcl-PHA

M. Zinn et al., Acta Biotechnol. 23, 309-316 (2003) Endotoxicity < 20 EU/device

B. Wampfler et al., 2010. Biomacromolecules, 11:2716-2723.

passage

over charcoal

OUTLINE

• SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS - TAILORING OF POLYMER ARCHITECTURE

• CONTROLLED DELIVERY SYSTEMS CONTAINING SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS

• SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS FOR BIOMATERIALS • SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS FOR COMPOSTABLE PACKAGES

structure

BIOMIXED ®

properties

HBA-CoA

n

n

HOC* H CH2CCoA

CH3

O

OC* H CH2C

O

CH3 H3C

O

O

-butyrolactone

enzyme initiator

PHB / poly(3-hydroxybutyric acid)

R.W. Lenz, Z. Jedliński, Macromol. Symposia 107, 149 (1996).

Z. Jedliński, M. Kowalczuk, P. Kurcok, G. Adamus, A. Matuszowicz, W. Sikorska,

R.A. Gross, J. Xu, R.W. Lenz, Macromolecules 29, 3773 (1996).

Novel polymerization of β-butyrolactone initiated by potassium naphthalenide in the presence of a crown ether or a cryptand

Zbigniew Jedlinski , Marek Kowalczuk , Wojciech Glowkowski , Janusz Grobelny , Michael Szwarc Macromolecules, 1991, 24 (2), pp 349–352

Poly[(R,S)-3-hydroxybutyrate]

anionic catalyst

e.g. ROM

supramolecular catalyst

e.g. ROM activated by 18crown6

no polymerization

O

CH3 On

PBL

BL

n

O

O

H3C

Activation of anionic species

Regioselectivity

Stereoselectivity

Propagation on carboxylate active centers

DEVELOPMENT OF THE BASES OF NEW

TECHNOLOGIES OF BIODEGRADABLE POLYMERS

MANUFACTURING BASED ON CLEAN PROCESSING OF

COAL

John W. Kramer, Emil B. Lobkovsky, and Geoffrey W. Coates

Org. Lett., 8, 2006, 3709-3712

CH3

O O OH

O R O O

R

O O OH

O R O O

R R

OR

O

Initiator: CH 3COOK / 18-Crownn-6

or CH

3COONBu

4

n

x

x

i) initiator

ii) H+

R = CH3-O-CH

2-; MOMPL

R = C2H

5-O-CH

2-; EOMPL

400 600 800 1000 1200 1400 1600 1800 2000

m/z

0

20

40

60

80

100

Rela

tive A

bundance

1243,5

1127,6 1359,5

1011,6

1475,4

1591,4 895,6

1707,4

779,6 1823,4

1939,4 719,6 663,6 487,3

New monomers

G. Adamus, M. Kowalczuk Biomacromolecules 2008

Copolyesters of

-butyrolactone with etoksymethyl--propiolactone

Random Copolyester

G. Adamus, Macromolecules, 2009, 42, 4547-4557

CH3

O O OH

O R O O

R

OC

2H

5-O-CH

2

O

OCH

3

O

n

2n-1

i)

ii) H+

R = CH3-; 3HBR = C2H5-O-CH2-; EOMPL

n+CH3COONBu4

OC

2H

5-O-CH

2

O

OCH

3

O

CH3

O O ONBu4

O CH3

O O

CH3

CH3

O O O

O CH3

O O

CH2

O

C2H

5

OH

CH2

O

O

C2H

5

m

i) n

CH3COONBu4

ii)

iii) H+

Step 1

Step 2

n-1

n m-1

Diblock Copolyester

ESI-MS/MS fragmentation spectrum

of [3-HB5/EMPL4 +Na]+

m/z 300 400 500 600 700 800 900 1000

0

10

20

30

40

50

60

70

80

90

100

Rela

tive

Ab

un

da

nce

973,2

887,2

715,2 801,2

1033,3

903,3 773,2

629,2 643,2

671,1 757,2 557,1 427,1 513,1

987,1 368,8

CH3

O O O O O O

O CH3

O CH3

O O CH2

O CH2

O CH2

O

O

C2H

5

O

C2H

5

O

C2H

5

O OH

CH2

O

C2H

5

OCH3

m/z 715 or m/z 629 or 543

m/z 903

m/z 801m/z 887m/z 973

-60 -146 -232

m/z 773

-130-260

+ Na

+

3

-390

m/z 643

-520

m/z 513

- 318 or -404 or- 490

ESI-MS/MS

fragmentation spektrum

of the molecular ion

at m/z 1033

selected from ESI-MS

spectrum of diblock

copolyester

Theoretical

fragmentation pathway

of the molecular ion at

m/z 1033 selected from

dibock copolyester

Diblock Copolyester

ESI-MS/MS fragmentation spectrum

ESI-MS/MS fragmentation spectrum of

the molecular ion at m/z 1033

selected from ESI-MS spectrum of

diblock copolyester

(m/z 1033)

(m/z 947) (m/z 903)

- 86 - 130

- 86 - 130

(m/z 861) (m/z 773)

- 86 - 130

-- 86- 86 -

- 86 -130- 86- 86 --- -

(m/z 817)

- 86 -- 86- 86 -130 - 130--- -

3HB5EMPL4+ Na

3HB4EMPL4 3HB5EMPL3

3HB2/EMPL4 3HB3EMPL33HB4EMPL2 3HB5EMPL

3HB3EMPL4 3HB4EMPL33HB5EMPL2

- 86

(m/z 775) (m/z 687)(m/z 731) (m/z 643)

- 86 - 130- 86 -- 86- 86 -130 - 130--- -

3HB2/EMPL3 3HB3EMPL2 3HB4EMPL

- 86

(m/z 689) (m/z 601)(m/z 645) (m/z 557)

3HB5

(m/z 513

- 130- 86

-60

BL5EMPL4

-86 -130

3HB4EMPL4 3HB5EMPL3

(m/z 887)

(m/z 973)

(m/z 843)

3HB3EMPL4 3HB4EMPL3 3HB5EMPL2

-86 -86 -130-130

(m/z 801) (m/z 757) (m/z 713)

3HB2EMPL4 3HB3EMPL3 3HB4EMPL2 3HB5EMPL

(m/z 715) (m/z 671) (m/z 627) (m/z 583)

-86 -86 -86 -130-130-130

3HBEMPL4

3HB2EMPL3 3HB3EMPL2 3HB4EMPL 3HB5

(m/z 629)(m/z 585) (m/z 541

(m/z 497) (m/z 453)

-86 -86 -86 -86 -130-130-130-130

3HBEMPL4

(m/z 1033)

(m/z 947) (m/z 903)

- 86 - 130

- 86 - 130

(m/z 861) (m/z 773)

- 86 - 130

-- 86- 86 -

- 86 -130- 86- 86 --- -

(m/z 817)

- 86 -- 86- 86 -130 - 130--- -

3HB5EMPL4+ Na

3HB4EMPL4 3HB5EMPL3

3HB2/EMPL4 3HB3EMPL33HB4EMPL2 3HB5EMPL

3HB3EMPL4 3HB4EMPL33HB5EMPL2

- 86

(m/z 775) (m/z 687)(m/z 731) (m/z 643)

- 86 - 130- 86 -- 86- 86 -130 - 130--- -

3HB2/EMPL3 3HB3EMPL2 3HB4EMPL

- 86

(m/z 689) (m/z 601)(m/z 645) (m/z 557)

3HB5

(m/z 513

- 130- 86

-60

BL5EMPL4

-86 -130

3HB4EMPL4 3HB5EMPL3

(m/z 887)

(m/z 973)

(m/z 843)

3HB3EMPL4 3HB4EMPL3 3HB5EMPL2

-86 -86 -130-130

(m/z 801) (m/z 757) (m/z 713)

3HB2EMPL4 3HB3EMPL3 3HB4EMPL2 3HB5EMPL

(m/z 715) (m/z 671) (m/z 627) (m/z 583)

-86 -86 -86 -130-130-130

3HBEMPL4

3HB2EMPL3 3HB3EMPL2 3HB4EMPL 3HB5

(m/z 629)(m/z 585) (m/z 541

(m/z 497) (m/z 453)

-86 -86 -86 -86 -130-130-130-130

3HBEMPL4

(m/z 1033)

(m/z 947) (m/z 903)

- 86 - 130- 86- 86 - 130

- 86 - 130- 86- 86 - 130

(m/z 861) (m/z 773)

- 86 - 130- 86- 86 - 130

-- 86- 86 -

- 86- 86 -130- 86- 86 --- -

(m/z 817)

- 86- 86 -- 86- 86 -130 - 130--- -

3HB5EMPL4+ Na

3HB4EMPL4 3HB5EMPL3

3HB2/EMPL4 3HB3EMPL33HB4EMPL2 3HB5EMPL

3HB3EMPL4 3HB4EMPL33HB5EMPL2

- 86

(m/z 775) (m/z 687)(m/z 731) (m/z 643)

- 86 - 130- 86- 86 - 130- 86- 86 -- 86- 86 -130 - 130--- -

3HB2/EMPL3 3HB3EMPL2 3HB4EMPL

- 86

(m/z 689) (m/z 601)(m/z 645) (m/z 557)

3HB5

(m/z 513

- 130- 86

-60

BL5EMPL4

-86 -130

3HB4EMPL4 3HB5EMPL3

(m/z 887)

(m/z 973)

(m/z 843)

3HB3EMPL4 3HB4EMPL3 3HB5EMPL2

-86 -86 -130-130

(m/z 801) (m/z 757) (m/z 713)

3HB2EMPL4 3HB3EMPL3 3HB4EMPL2 3HB5EMPL

(m/z 715) (m/z 671) (m/z 627) (m/z 583)

-86 -86 -86 -130-130-130

3HBEMPL4

3HB2EMPL3 3HB3EMPL2 3HB4EMPL 3HB5

(m/z 629)(m/z 585) (m/z 541

(m/z 497) (m/z 453)

-86 -86 -86 -86 -130-130-130-130

3HBEMPL4

300 400 500 600 700 800 900 1000m/z

0

10

20

30

40

50

60

70

80

90

100

Rela

tive A

bundance

973,2

843,2

903,3757,2

887,2 1033,2

817,2671,2

687,2 801,2

601,2

541,1

497,1455,2325,0 411,0 974,1

947,3

861,2

773,2

713,2

300 400 500 600 700 800 900 1000m/z

0

10

20

30

40

50

60

70

80

90

100

Rela

tive A

bundance

973,2

843,2

903,3757,2

887,2 1033,2

817,2671,2

687,2 801,2

601,2

541,1

497,1455,2325,0 411,0 974,1

947,3

861,2

773,2

713,2

Theoretical fragmentation

pathway of the molecular ion at

m/z 1033 of random copolyester

of [3-HB5/EMPL4 +Na]+

O C

O

O

CH

O

CH

2

C

O

O

O C

O

O

CH

O

CH

2

C

O

O

O C

O

O

CH

O

CH

2

C

O

O

CH

CH

2

C

O

O

CH3

O C

CH3

O

-butyrlactone

-butoxymethylpropiolactone -phenoxymethylpropiolactone -benzoxymethylpropiolactone

CH3

O O OH

O R O O

R

OCH

3

O

O C

O

O2n-1

i)

ii) H+n+

CH3COONBu4

x

400 600 800 1000 1200 1400 1600 1800 m/z

0

20

40

60

80

100

Re

lative

Ab

un

da

nce

1136.6

1221.6 1307.6

1049.5 1393.6 963.5

877.6 1500.5

857.5

1585.4 772.4

1605.4

1778.3 686.2 1863.2

599.6 367.5

950 1000 1050 1100 m/z

0

20

40

60

80

100

Rela

tive A

bundance 1049.6

963.6 1050.7

964.7 943.5 1029.5

1069.5

983.7

1090.3 1010.1 924.5

86 Da

86 Da

20Da

20Da

20Da

HB10

HB8BHB

HB6BHB2

C H

C H 2

C

O

O

C H 3

C H

O

C H 2

C

O

O

x = or x=

HB; M = 86 BHB; M = 192

950 1000 1050 1100 0

20

40

60

80

100

Rela

tive A

bundance

1049.6 963.6

1050.7

964.7 943.5 1029.5

1069.5

983.7

1090.3 1010.1 924.5

86 Da

86 Da

20Da

20Da

20Da

HB10

HB8BzHB

HB6BzHB2

HB9BzHB

HB8BzHB

-60 Da

m/z

C H

C H 2

C

O

O

C H 3

C H

O

C H 2

C

O

O

x = or x=

HB; Mcz 86 BzHB; Mcz 192

300 400 500 600 700 800 900

0

20

40

60

80

100

Re

lative

Ab

un

da

nce

963.2

903.2

817.1

731.1 877.2

645.1

791.2 705.1 559.1 599.2

513.1 771.1

453.1 426.9 366.9 -192 Da

-86Da

-86 Da

OUTLINE

CONTROLLED DELIVERY SYSTEMS CONTAINING SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS

86 n-1

- K+

S CH3

CH3

CH

O O

N

H H

O

C N

O

HH H

CH CH2 C

OCH3

O CH

CH3

CH2 C

O

OK

_

β - Lactame antibiotics

G. Adamus, M. Kowalczuk,

Rapid Commun. Mass Spectrom., 14, 195, 2000

400 600 800 1000 1200 1400

m/z

20

40

60

80

100

Re

lati

ve

Ab

un

da

nc

e 849.1

935.1 763.2

1021.1

677.2 1107.0

1193.0 591.2

505.2 1279.0

419.2 1364.9

Oligo(3-hydroxybutanoate) conjugates with acetylsalicylic acid

European Journal of Medicinal Chemistry 43 (2008) 1785

Cytotoxicity of different doses of ASA, ASA bound to oligomer (ASAeOHB)

and ASA mixed with oligomer

Synthesis and antiproliferative properties of ibuprofen–oligo(3-hydroxybutyrate) conjugates

B. Zawidlak-Węgrzyńska, M. Kawalec, I. Bosek, M. Łuczyk-Juzwa, G. Adamus, A. Rusin, P. Filipczak,

M. Głowala-Kosińska, K. Wolańska, Z. Krawczyk, P. Kurcok European Journal of Medicinal Chemistry 45 (2010) 1833

O

O

CH3

O O

O CH3 O

HS

S

n

ni) inicjator

ii) H+

inicjator:

rozpuszczalnik: DMSO

O-

O

S

S K+

Immobilization of active cosmetic components

Lipoic acid

OUTLINE

SYNTHETIC ANALOGUES OF ALIPHATIC BIOPOLYESTERS FOR

BIOMATERIALS

structure

properties

BLENDS

BIOMIXED ®

a-PHB/PLGL

Blends

2007-01-30_13;03;40_Y1_01.vdt: Refractive Index

-6.00

5.67

17.35

29.02

40.70

52.37

64.05

75.72

87.39

Re

fra

ctiv

e In

de

x (

mV

)

Retention Volume (mL)

0.10 2.24 4.37 6.51 8.64 10.78 12.91 15.05 17.19 19.32 21.46 23.59 25.73 27.86 30.00

Mn 82 600

Mw/Mn 1.2

a-PHB

BIOMIXED ®

structure

properties

BLENDS

MISCIBILITY OF BLENDS OF LACTIDE HOMO AND

COPOLYMERS WITH ATACTIC POLY(3-HYDROXYBUTYRATE)

Patent RP P-320208

M. Focarete, M. Scandola,

P. Dobrzynski, M. Kowalczuk,

Macromolecules 2002, 35, 8472

M. Gazzano, M. Focarete,

Ch. Riekel, M. Scandola,

Biomacromolecules 2004, 5, 553

Fibers prepared from a-PHB/PLGL

blend

Artificial 3D scaffolds aPHB-10k/nPHB -80/20

SEM photographs of fibers obtained after electrospinning

of the solutions more viscous (left) less viscous (middle)

and the least viscous (right) forming particles due to

electrospray

a-PHB

COATINGS OF ARTIFICIAL HEART EXTERNAL PROTESIS

Obraz żywych oraz nekrotycznych fibroblastów w hodowli z materiałem a-PHB C01856 - komórki nekrotyczne wykazują czerwoną fluorescencję (barwienie jodkiem propidyny). Powiększenie 10x a-PHB - 17,3 ± 2,08 komórek POLVAD-IMPL

OH OH

R O

n

CHCl3/H

2O

O O O

R O R O R O

m

K+

OCH

3

O

H+

O O O O

R O R O R O CH3

O

m

H

y

PHA

KOH/18-crown-6

/ 18-crown-6

(i) y

(R,S) BL

(ii)

PHA-block-(a-PHB)

Novel block copolymers of atactic PHB with natural PHA

BIOCOP ®

European Polymer Journal 48 (2012) 621–631

Controlled depolymerization of aliphatic polyesters from natural resources

R O

OHOH

n

O O

O O

p

OH

O O

O O

r

OH O OOH

O O R

m

O

O

OHs

OH

transesteryfication/glycol

hydrolysis/acid

hydrolysis/basic

reduction/sodium borohydride

PHA-block-(a-PHB) COPOLYMERS

BIOCOP ®

H

y

(PHA)-block-(a-PHB)

yO

O

H3C ( )R, S BL

CHCH

CO

CHCH2

CO

CHCH2

CO

-

RR O O OR

x

/18-crown-6K+

CHCH

CO

CHCH2

CO

CHCH2

C

RR O O OR

x

OCH

CH2C

O

OCH3

PHB: R = CH3

PHBV: R = CH3, C2H5

PHO: R = C3H7, C5H11,C7H15

H+

(i) (ii)

Average number of bond cleavages per

polymer molecule (N) as a

function of hydrolytic degradation time:

natural PHB; PHB-block-(a-PHB)

copolymer; plain a-PHB

Synthesis and processing

LABORATORY OF BIODEGRADABLE MATERIALS

38 38

GENERAL VIEW OF PLA FILM

EXTRUSION STAND

a) Extrusion of sheet film.

b) IR film thickness measurement.

c) Thermoforming.

39 39

THE PLA FILM EXTRUSION STAND

40 40

SPECIAL TESTING STAND FOR VACUUM

THERMOFORMING OF PLA

Nr umowy POIG.01.03.01-00-018/08-00

Zdjęcia makroskopowe rolek folii

PLA i PLA/a-PHB

PLA/a-PHB

MARGEN

COMPOSTING

The first advice on rational composting in Poland: “The Farmstead” by Anzelm Gostowski, 1563

http://naturemill.com/products.html

1644

Innovative value chain development for sustainable plastics in Central Europe

Case study 2B

SYSTEMIC APPROACH FOR SUSTAINABLE

PRODUCTION FOR BIOPLASTICS

COMPOSTING

Innovative value chain development for sustainable plastics in Central Europe

Case study 2B

Schematic diagram of the organic

recycling of packaging materials

Innovative value chain development for sustainable plastics in Central Europe

http://www.modellprojekt-kassel.de/eng/downloads/kassel-project_brochure.pdf

Case study 2B Case study 2B

Innovative value chain development for sustainable plastics in Central Europe

Case study 2B

Innovative value chain development for sustainable plastics in Central Europe

Pile composting plant of organic wastes as well as other composting

plant (with preliminary compost in containers and maturation of

compost on piles) started in 1999 in Zabrze

The pile consisted of 40% leaves,

30% branches and 30% grass. The containers consisted

approximately of 40%

kitchen organic waste, 20%

leaves, 20% branches and

20% grass.

Case study 2B

Innovative value chain development for sustainable plastics in Central Europe

Case study 2B

Diagram of composting process in container system

Degradation under industrial composting conditions

Innovative value chain development for sustainable plastics in Central Europe

Cage with the samples

The cage with polymeric materials

was placed in composting pile at

the depth of one meter under the

surface

Case study 2B

Digital photographs of samples before degradation, after 7 and 21 days degradation

in container system

PLA

0 7 21 days

98/2

0 7 21 days

Degradation under industrial composting conditions

Innovative value chain development for sustainable plastics in Central Europe

Case study 2B

Growth of (a) barley and (b) cress in soil after 183 days of degradation of:

(1) a-PHB/n-PHB/ - and n-PHB and (2) a-PHB/PLLA - and PLLA films

Growth of cress and barley measured as percent

of dry weight against the control

Innovative value chain development for sustainable plastics in Central Europe

Case study 1B

Testing of markers for easy identification of

biodegradable plastics in the waste stream

LABORATORY OF BIODEGRADABLE MATERIALS

III : Piotr Kurcok, Michał Sobota, Grażyna Adamus, Lilianna Miga, Michał Kwiecień II : Wanda Sikorska, Joanna Rydz, Łukasz Rudnick, Magdalena Maksymiak I : Marta Musioł, Iwona Romanowska, Katarzyna Wolna-Stypka, Barbara Zawidlak

THANK YOU FOR YOUR KIND ATTENTION