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A collaborative MOOC-Project by: A collaborative MOOC-Project by: part of the part of the Sustainable biopolymers Production, applications and future directions Lars M. Blank A collaborative MOOC-Project by: part of the Perspectives


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A collaborative MOOC-Project by: part of the A collaborative MOOC-Project by: part of the

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A collaborative MOOC-Project by: part of the

Sustainable biopolymers

Production, applications and future directions

Lars M. Blank

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A collaborative MOOC-Project by: part of the

Lars M. Blank & Nick Wierckx

Perspectives concerning biopolymers

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Worldwide polymer/plastic production 2015 2017 2010

Adapted from Jambeck et al., Science 2015; Data origin: 1 Plastics Europe, „Plastics – The Facts 2013“; 2 Cozar et al., 2014; Erikson et als., 2014; 3 European Bioplastics, nova-Institute 2015; 4 Plastics Europe, „Plastics – The Facts 2016“; 5 European Bioplastics, nova-Institute 2017

6,350-245,000 t 2

estimated plastic waste floating at the ocean surface

393 Mt

global polymer production3

275 Mt

total plastic waste1

31.9 Mt

coastal mismanaged plastic waste

2.05 Mt

bio-plastic production5

322 Mt

global plastic production4

8 Mt

plastic waste goes into the see

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Sustainable biopolymers

  • up-cycling of plastic waste

bacteria degrading fossil-based plastic and producing biodegradable plastic from monomers

  • new bio-based polymers with higher performance and value

e.g. PLA and PHA

  • bio-based drop-in polymers

à create “bio-versions” of existing monomers and polymers e.g. bio-PET and bio-PE

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From 30 % to 100 % bio-based PET

  • currently, 13 – 30 % bio-based PET available
  • main bottleneck: bio-TPA

source: Coca Cola Company, Danone

Nestlé & Danone

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A collaborative MOOC-Project by: part of the

Alternative: PEF – polyethylene furanoate

terephthalic acid (TPA) O O HO OH

  • based on 2,5-furandicarboxylic acid (FDCA)
  • bio-based alternative to terephthalate
  • PEF has better barrier properties than PET
  • compatible with PET recycling (<5%)

FDCA O O HO OH O

www.synvina.com/products/pef/ source: Corbion, Synvina

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A collaborative MOOC-Project by: part of the

How did we get to bio-based FDCA?

FDCA

?

  • via 5-hydroxymethyl furfural (HMF)
  • product of thermochemical treatment of cellulose-rich biomass
  • common inhibitor in lignocellulosic hydrolysate

HMF O O HO OH O O HO O

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A collaborative MOOC-Project by: part of the

Search for HMF-degrading bacteria

  • enrichment cultures
  • isolate HMF-degrading bacteria from nature
  • identified isolated bacteria

→ Cupriavidus basilensis HMF14

Wierckx et al. Microbial Biotechnol. 3:336-343. Patent WO2011026906

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Characterized HMF degradation pathway

  • identified hmf genes
  • characterized HMF pathway

→ intermediate: FDCA!

  • HMF oxidase HmfH enables FDCA production

Koopman, Wierckx et al. Bioresource Technol. 101:6291-6296 Patent WO2011026913; WO2012064195

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Whole-cell biocatalyst

HmfH HMF HMF acid FDCA HMF hmfH FDCA endogenous dehydrogenases HmfH

  • P. putida S12
  • whole-cell biocatalyst
  • cofactor integration
  • easy enzyme production
  • solvent-tolerant Pseudomonas putida
  • reactive substrate
  • H2O2 by-product
  • versatile metabolism
  • faster conversion due to dehydrogenases

O O HO OH O O O HO OH O O HO O O HO O O HO OH O

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A collaborative MOOC-Project by: part of the

Fed-batch conversion of HMF

5 10 15 20 25 30 35 30 60 90 120 150 biomass, HMF acid, FDCA (g/l) time (h)

FDCA biomass HMF acid

start HMF feed decrease C- source feed decrease HMF feed increase C- source feed

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Purify the product

boil centrifuge pH→0.5 extraction with THF biomass water Impurities impurities 99.4% pure FDCA

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Timeline: from bacterial isolation to purified product

  • May 2007: collect soil and water samples
  • Oct 2007: identified Cupriavidus basilensis
  • Jun 2008: characterized HMF degradation route
  • Nov 2008: constructed FDCA production host
  • Patent applications
  • Sep 2009: produced 1st 10 grams of pure FDCA
  • Mar 2011: produced 1st 500 grams of pure FDCA
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A collaborative MOOC-Project by: part of the A collaborative MOOC-Project by: part of the