Available online at www.sciencedirect.
com
ScienceDirect
Biotechnological upcycling of plastic waste and other
non-conventional feedstocks in a circular economy
Lars Mathias Blank1, Tanja Narancic2, Jörg Mampel3, Till Tiso1 and
Kevin O’Connor2
The envisaged circular economy requires absolute carbon
efficiency and in the long run abstinence from fossil feedstocks,
and integration of industrial production with end-of-life waste
management. Non-conventional feedstocks arising from
industrial production and societal consumption such as CO2
and plastic waste may soon enable manufacture of multiple
products from simple bulk chemicals to pharmaceuticals using
biotechnology. The change to these feedstocks could be faster
than expected by many, especially if the true cost, including the
carbon footprint of products, is considered. The efficiency of
biotechnological processes can be improved through
metabolic engineering, which can help fulfill the promises of the
Paris agreement.
Addresses
1
iAMB – Institute of Applied Microbiology, ABBt – Aachen Biology and
Biotechnology, RWTH Aachen University, Worringer Weg 1,
52074 Aachen, Germany
2
BEACON SFI Bioeconomy Research Centre and School of
Biomolecular and Biomedical Science, University College Dublin,
Belfield, Dublin 4, Ireland
3
BRAIN AG, Darmstädter Str. 34-36, 64673 Zwingenberg, Germany
Corresponding author: Blank, Lars Mathias (lars.blank@rwth-aachen.de)
Current Opinion in Biotechnology 2020, 62:212–219
This review comes from a themed issue on Energy biotechnology
Edited by Joe Shaw and Kirsten Benjamin
https://doi.org/10.1016/j.copbio.2019.11.011
0958-1669/ã 2019 Published by Elsevier Ltd.
Introduction
The envisaged circular economy requires the utilization
of every side stream to minimize waste and ultimately
CO2 formation. The chemical industry today relies on
crude oil and other fossil resources as carbon (in
2018 about 880 Mt [1]) and energy feedstocks for about
90% of its products. By comparison, in crop year 2016/
2017 2.2 billion tons of grain has been produced [2]. In the
EU in crop year 2014/2015 approximately 24% of the
grain was used for direct human consumption, while
about 62% was produced for animal feed [3]. Strong
market growth makes the chemical industry the largest
Current Opinion in Biotechnology 2020, 62:212–219
driver for future increase in global fossil oil demand
already by the year 2030 [4]. Kätelhön et al. presented a
CO2-only scenario for the chemical industry, replacing all
current fossil feedstocks. While such a scenario is in
principle feasible, the required CO2-free produced elec-
trons sum-up to a staggering 18.1 PWh, representing 55%
of the world’s electricity generated in 2030 [5]. The likely
scenario will be a chemical industry shifting to a mixture
of carbon sources, including for biotechnological pro-
cesses both conventional (e.g. starch, molasses) and
non-conventional feedstocks (e.g., CO2, methane, glyc-
erol, agro and urban waste streams) [6].
While the search for even cheaper feedstocks is at least a
century old (i.e. ABE fermentation was switched from
starch to cheaper molasses in the 1920’s), the minimization
of the carbon footprint in a circular economy adds another
motivation. Non-conventional feedstocks have a battery of
constraints to fulfill, including price and its fluctuation,
availability, and ease of use, to name a few. Methanol (and
methane) was prominently worked on in the 1970’s and 80’s
and is now complemented by formate [7] as a carbon source
that can be readily synthesized from CO2 via (electro)
chemical catalysis [8]. Acetate can be synthesized by aceto-
gens from CO2 and H2 [9], in combination with glucose at
rates of about 30 mmol/gCDW/h [10], while the direct (elec-
tro)chemical route might also be feasible in the future.
While glycerol is discussed for more than a decade as a
feedstock originating from biodiesel production [11], also
new plastic pyrolysis oils [12] and plastic hydrolysates [13]
are complementing the potential substrate range but their
usage still requires further development. With more than
350 million tons per year of newly produced plastic [14], a
market growth rate of 5+%, and a staggering 4.8 billion tons
of plastic in landfills [15], the possibilities for these non-
conventional feedstocks are excellent, especially when
considering the combination of production with end-of-
life management [16].
These side and waste streams can only be considered as
non-conventional feedstocks because of the bow-tie
structure of many microbial metabolic networks (see
graphical abstract). A multitude of catabolic pathways
exist that merge in the central carbon metabolism,
defined by the reactions synthesizing the 12 metabolites
required for biomass synthesis [17]. These anabolic
reactions can be exploited for biocatalysis of valuable
products or extended by new-to-nature synthesis
pathways [18,19].
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 Upcycling of non-conventional feedstocks Blank et al. 213

Here, we benchmark a collection of non-conventional
feedstocks against sugars regarding their theoretical yield,
review the fast upcoming ‘plastic waste to plastic value’
biotech community, and elucidate the possibilities of
modular utilization of CO2 in combination with other
non-conventional feedstocks.
Non-conventional feedstocks for maximizing
product yields
While sugars from starch and molasses are the preferred
carbon sources in industry, the theoretical yield of Cmol
(mole of carbon of the metabolite in question, for exam-
ple, 1 mol glucose has 6 Cmol) metabolite per Cmol
feedstock is higher for most non-conventional feedstocks
(Table 1). Only terephthalic acid from plastic waste and
acetate display generally lower yields. To cautiously
generalize, highly reduced feedstocks (e.g., dodecane,
methanol, ethanol, with degrees of reduction [20] of six
and higher) have higher yields, compared to more oxi-
dized feedstocks like terephthalic acid and acetate
(degrees of reduction of 3.75 and 4, respectively). The
extra electrons in the highly reduced feedstocks are
available for CO2 reduction to ultimately funnel addi-
tional carbon into products, for example via the anaplero-
tic activity of the pyruvate carboxylase into the reductive
TCA cycle. This additional carbon guarantees higher
yields for TCA cycle intermediates such as oxaloacetate,
oxoglutarate, and succinyl-CoA. Examples are malate
production from glycerol at a concentration of about
200 g/L [21] and succinate production with a yield of
0.91 molsuccinate/molglycerol [22]. This high theoretical
yield is also exploited for 2,3-butanediol production from
glycerol using cyanobacteria [23] and 1,2-propanediol
production by Saccharomyces cerevisiae [24]. Yaguchi
et al. review engineering approaches to enable yeasts
for growth on feedstocks such as acetate, aromatics,
and fats [25], while the uptake of C1 feedstocks using
bacteria has been reviewed by Clomburg et al. [6].
Efficient metabolization of ethylene glycol from plastic
waste has recently been reported with Pseudomonas putida
[26,27]. In industry, carbon mixtures are often applied,
for example, in wheat straw-based processes. Well-
designed combinations of non-conventional feedstocks
allow even higher theoretical yields, while imposing extra
challenges such as sequential feedstock use [28].
To highlight the importance of pathway use for the
theoretical yield, here the ‘isomerase pathway’ and the
‘Weimberg pathway’ for xylose catabolism are exemplar-
ily compared. Indeed, while the feedstock is identical, the
theoretical yields calculated differ substantially (Table 1).
The Weimberg pathway for xylose utilization was intro-
duced into a number of microorganisms, including Escher-
ichia coli [29,30], S. cerevisiae [31], and P. putida [32], while
being naturally present in other pseudomonads [33].
Examples for synthetic pathways with improved stoichi-
ometry are 2,4-dihydroxybutyric acid production from
homoserine [34], acetyl-CoA-based biocatalysis via the
designed EP-bifido pathway [35], and novel routes for
formate [36] and CO2 utilization [19,37]. Tools for the
prediction of enzyme activities in these synthetic
approaches are truly required [38].
Upcycling of plastic waste
The ‘take-make-dispose’ culture associated in particular
with single-use plastics poses serious challenges for sus-
tainability given the undesirable consequences of post-
consumer plastic. Globally, 80% of all plastic produced,

Table 1

Theoretical yields determined by flux balance analysis. The yields are expressed in Cmol metabolite per Cmol feedstock and were
calculated using the genome scale model iJN1411 of P. putida [39]. The chosen molecules represent the 12 metabolites of central carbon
metabolism that are required for the synthesis of all biomass constituents, and hence represent proxies for the multitude of potential
upcycling products (each column corresponds to one metabolite). The feedstocks (each row corresponds to one feedstock) comprise the
benchmarks glucose and xylose, glycerol as side-product from biodiesel production, ethanol and acetate, which are envisioned to be
synthesized from CO2 with acetogens, and methanol as example for a C1 feedstock via (electro)chemical catalysis also from
CO2. Ethylene glycol and terephthalic acid originate from PET plastic waste by (bio)chemical depolymerisation, while via pyrolysis
polyolefins can be converted into oils (here dodecane as proxy). The shading represents the value of the yield: the darker the grey, the
higher the yield

G6P F6P R5P E4P G3P 2PG PEP PYR AcCoA OAA OGA SucCoA mean
Glucose 0.90 0.90 0.89 0.86 0.81 0.98 0.98 1.00 0.67 1.33 0.83 0.79 0.91
XyloseIsomerase 1.00 1.00 1.00 0.97 0.91 1.00 1.00 1.00 0.67 1.33 0.83 0.82 0.98
XyloseWeimberg 0.60 0.60 0.60 0.60 0.60 0.60 0.60 0.60 0.40 0.80 1.00 0.80 0.65
Glycerol 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 0.67 1.33 0.83 0.81 1.00
Ethanol 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 1.00 1.00 0.83 1.00 0.82
Acetate 0.67 0.67 0.67 0.67 0.67 0.75 0.75 0.75 0.86 1.00 0.83 0.89 0.77
Methanol 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 0.67 1.33 0.83 0.77 0.97
Ethylene glycol 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.50 1.00 0.63 0.67 0.73
Terephthalic acid 0.54 0.54 0.54 0.54 0.54 0.56 0.56 0.56 0.50 0.75 0.63 0.72 0.58
Dodecane 0.75 0.75 0.75 0.75 0.75 0.75 0.75 0.75 1.00 1.00 0.83 1.00 0.82

G6P – glucose-6-phosphate; G3P – glyceraldehyde-3-phosphate; R5P – ribulose-5-phosphate; E4P – erythrose-4-phosphate; 2PG – 2-phospho-
glycerate; AcCoA – acetyl-CoA; OAA – oxaloacetate; OGA – oxoglutarate; SucCoA – succinyl-CoA; PYR – pyruvate; F6P – fructose-6-phosphate;
PEP – phospho-enol-pyruvate.

www.sciencedirect.com Current Opinion in Biotechnology 2020, 62:212–219

214 Energy biotechnology
roughly equivalent to 3.5 billion tons of carbon (based on
approximately 60% carbon content of plastics), has not
been re-used by society due to the lack of recycling and
re-use technologies for these materials [15]. The circu-
lar economy of plastics focuses on the retention of value in
the material cycle through recycling and re-use of plastic
materials. However, most currently used plastics are
not efficiently recycled, leading to environmental pro-
blems [40]. There is a loss of 95% of the material value of
plastics after just one use, amounting up to s110 billion
being lost to the economy each year [41]. A current gap in
the circular economy concept with respect to plastics is
the role of biodegradable plastics. Upcycling of plastic
waste to biodegradable plastic and chemicals as a means
of valorization of post-consumer plastic should also be
part of this new thinking.
Recently, partially deconstructed petroleum-derived poly-
ethylene terephthalate (PET) was combined with renewably
sourceable monomers (fumarate, malate, and muconate) to
produce high-value fiber-reinforced plastics [42]. This inno-
vative approach in plastic upcycling offers alternatives to
previously used approaches in partial deconstruction of PET
and consequent use in the production of epoxy resins,
polyurethanes, and unsaturated polyesters [42].
We have reported on the combination of pyrolysis and
microbiology to convert non-degradable plastics such as
PET, polyethylene (PE), and polystyrene (PS) into
biodegradable counterparts, namely polyhydroxyalkano-
ates (PHA), offering an unconventional route to
connect technical and biological parts of the circular
economy [12,43–45].
Enzymatic hydrolysis of plastics has great potential
given that there are many enzymes that degrade complex
polymers in nature [13]. In 2016, Yoshida et al. reported a
newly isolated bacterium, Ideonella sakaiensis 201-F6, with
the unusual ability to degrade PET and assimilate its
monomers [46]. While demonstrating the potential of
nature to degrade plastics, the capacity of this bacterium
for PET degradation is limited. However, this has opened
the door for the characterization and engineering of the I.
sakaiensis key enzymes involved in PET degradation,
designated as PETase and MHETase [47,48,49] with
great potential for further engineering. The main chal-
lenge of the enzymatic hydrolysis of PET is the need for
higher incubation temperatures due to the high glass
transition temperature, leading to physical aging of
PET [50]. Efforts such as rational protein engineering
of I. sakaiensis for more efficient PET degradation under
mild conditions, that is, at 40 C are paramount for achiev-
ing biodegradation of PET at biologically relevant
conditions [49].
The generated monomers terephthalic acid and ethylene
glycol from PET can be used as feedstock for
Current Opinion in Biotechnology 2020, 62:212–219
biodegradable plastics and as chiral building blocks
[26,44,51], exploiting the bow-tie structure of microbial
metabolism. The use of enzymes for biodegradation and
microbes or enzymes for product formation would repre-
sent a completely biological recycling of plastics in a
circular economy [52]. The integration of hydrolytic
enzymes into a microbial biosynthesis chassis would
result in a custom microbial platform capable of convert-
ing carbon trapped in plastic into valuable products using
a single organism [53,54].
The environmental impact of biotechnological products
arising from plastic recycling must be considered as early
as possible in the design of those technologies [16]. Our
investigation [16] clearly shows the need to manage
rather than release the biodegradable plastics into the
environment, and that the management of these plastics
will enable much more efficient access to the carbon
trapped in these biodegradable polymers providing for
far greater resource/carbon efficiency. The learnings from
enzymatic hydrolysis of fossil-based plastics can also be
applied to more readily biodegradable plastics like poly-
lactic acid (PLA), so that they can be used as carbon
sources for biotechnology – in the envisaged circular
economy.
Flexibility in feedstock use
Upcycling of abundant CO2 vitally depends on
abundance of renewable energy
Anthropogenic production of CO2 outnumbers the
demand of a future carbon-reduced chemical industry
by a factor of 10 [5]. Together with some 2–3000 Gt of
atmospheric CO2 and direct air capture technologies, it is
clear that atmospheric CO2 as feedstock enables almost
unlimited scalability for commodities. However, in order
to be neutral or positive in Life Cycle Assessment (LCA),
the chemical reduction of CO2 has to rely on renewable
energy sources (green energy), a challenging task. Molec-
ular hydrogen (H2) is of special interest and technologies
under development use even seawater as substrate
for electrolysis [55]. Moreover, H2 can be stored and
transported. Energy storage enables balancing of the
immanently fluctuating green energy supply, a key
requirement toward a carbon-neutral circular economy.
Three major modes for upcycling of CO2 exist: (electro)
chemical CO2 reduction, combined chemical–biological
processes, and biological CO2 fixation. Co-polymerization
of CO2 with highly energetic molecules is frequently used
[56], but only the direct electrochemical reduction of CO2
enables the synthesis of fully CO2-derived polymers.
Whereas selective carbon chain elongation is a major
challenge in organic chemistry, cells synthesize mole-
cules with carbon chain lengths up to C40, but are notori-
ously slow in the activation of CO2, with some exceptions
[57]. Combining chemical CO2 reduction with biological
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 Upcycling of non-conventional feedstocks Blank et al. 215

chain elongation is thus an attractive option [58]. Pioneer-
ing concepts of coupled chemical–biological CO2 conver-
sions were recently introduced [59] and lately gained
momentum using formate as a versatile platform mole-
cule [7]. A variety of naturally evolved microorganisms
use formate as sole source of carbon and energy, but
synthetic biologists have started to redesign central met-
abolic pathways in more robust platform organisms to
convert formate into value-added products [36]. Besides
formate, methanol and carbon monoxide (CO) are further
prime platform molecules that fit into coupled chemical–
biolocigal CO2 upcycling concepts [60,61,62,63].
The third option to harness CO2 as feedstock relies
exclusively on microorganisms that either partially or
completely incorporate CO2 in biomass and fermentation
products via carboxylation reactions or autotrophic
metabolism, respectively. Autotrophic organisms employ
three different modes for CO2 activation. While the use of
light by phototrophic microorganisms or the use of
reduced chemical compounds (e.g. H2) by chemolithoau-
totrophic bacteria is well established, the direct use of
electricity by exoelectrogenic bacteria (microbial electro-
synthesis, MES) is a novel concept, which is still under
debate [64,65]. Chemolithoautotrophic bacteria gain
reductive energy from gaseous substrates and produce
short chain alcohols and acids as products, by native or
engineered pathways [66]. Because of the sensitivity of
the organisms toward oxygen, ATP-intensive production
pathways are constrained. The metabolic flexibility of
these organisms allows mixotrophic growth that can
relieve some of these constraints [67], but aerobic

Figure 1

anoxic conditions

(a)
green energy
acetogens
O2 oxic conditions

homoacetogens

(b)

A. woodii E. coli
Y. lipolytica

- 400

- 200
formate
0
methanol
(c) ethanol (d) +200
Δ 1.23 V
ethylene glycol +400

lactate +600

C6 sugars +800

Current Opinion in Biotechnology

Upcycling of CO2 via the versatile acetate-platform.

Renewable energy (green energy) provides CO2-free electrons to generate the reductive power ([H]) that is needed to activate CO2. (a) Acetogens
convert CO2 or CO into one major product (P1) along with side products either via natural or recombinant pathways. (b) Homoacetogenic bacteria
like Acetobacterium woodii convert CO2 into acetate as the single product, achieving high carbon and Faradaic efficiency (i.e., efficiency of
electron transfer into products). (c) A. woodii also employs homoacetogenesis to metabolize via its bow-tie structure metabolism a variety of
reduced carbon species to acetate as single product, thus enabling feedstock flexibility which is economically highly attractive. Acetate is further
metabolized into value-added products under oxic conditions by a second heterotrophic microorganism (MO2) of choice (natural or recombinant)
that uses oxygen as terminal electron acceptor. A variety of acetate assimilating organisms (bacteria, yeast, fungi) produce a broad variety of
products (P1, P2 . . . ) via natural or recombinant pathways. (d) The availability of molecular oxygen as terminal electron acceptor harnesses the
full energetic potential (1.23 V) of the reductive energy thereby enabling the biosynthesis of products that are not amenable by purely anaerobic
(i.e., in absence of exogenous electron acceptors) processes.

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216 Energy biotechnology
chemolithoautotrophic bacteria overcome energetic lim-
itations more effectively. The synthesis of highly reduced
products like alkanes or terpenes [68] with engineered
Cupriavidus necator stains demonstrates the potential of
this concept.
Acetate as flexible platform for highly efficient
valorization of CO2
Acetogens are a diverse group of anaerobic bacteria that
rely on the Wood–Ljungdahl pathway (WLP) for the
formation of acetyl-CoA, a universal metabolic hub for
biosynthesis. The WLP being the only pathway that
couples energy conservation with CO2-fixation, is the
most energy efficient of the seven natural and two syn-
thetic CO2-fixation pathways [69] and presumably also
is present in aerobic bacteria [70]. Acetobacterium woodii is
the model organism for a group of acetogens that form
acetate as the exclusive product of CO2 reduction gaining
only 0.3 mol of ATP from the reduction of 2 mol of CO2
[71]. Consequently, > 94% of CO2 is converted into
acetate. Only under oxic conditions acetate can serve
as sole source for carbon and energy and is metabolized
via the citric acid cycle (to gain energy) or the glyoxylate
shunt (to form biomass). The interplay of these
two pathways and the respiratory chain enables the bio-
synthesis of a broad range of valuable bio-products (Sup-
plementary Table). Several such concepts have been
demonstrated recently for the production of various com-
pounds, including alkanes and triacylglycerides (Supple-
mentary Table). Feedstock flexibility is a specific advan-
tage of acetate-based upcycling concepts (Figure 1).
Homoacetogenic bacteria metabolize formate, methanol,
ethanol, and glycols as well as sugars, to form acetate as
the sole product. Notably, acetogens can convert 1 mol of
glucose into 3 mols of acetate, which matches the carbon-
efficiency of the recently designed synthetic NOG-bifido
[72] and EP-bifido [35] pathways. Moreover, acetate is an
abundant feedstock aerobically produced from glucose
(by acetic acid bacteria) or as a by-product of hydrolysis
of lignocellulosic biomass. Recently, a ‘cyborgian
bacterium’ (i.e. Moorella thermoacetica decorated with cad-
mium sulfide nanoparticles) was introduced that provided
proof of concept for the photosynthetic WLP-mediated
formation of acetate – energetically the most appealing
solution and a highly stimulating concept for future CO2
upcycling efforts [73,74].
Conclusions
Non-conventional feedstocks are being used and will
increasingly contribute to the development of the envis-
aged circular economy. While the bow-tie metabolism of
microbes allows a myriad of feedstocks, exploited for
decades in such engineered environments as waste-water
treatment and biogas synthesis, the here reviewed dedi-
cated use of non-conventional feedstocks with engi-
neered microbes has only few industrially implemented
examples until now. The challenges range from basic
Current Opinion in Biotechnology 2020, 62:212–219
biochemistry, as details of degradation pathways are
unknown, via classic microbiology, to the metabolic engi-
neering challenges of TRY (titer, rate, and yield), and
further to markets. The transfer to market is a true
challenge for industry, as cost, performance, and risk of
a new technology is often unfavorable when compared to
depreciated chemical plants, explaining the gap between
the many academically driven proof-of-concept projects
and the limited commercial exploitation. To complain
about the oil price is not helpful, while the price fluctua-
tions of many non-conventional feedstocks are directly or
indirectly influenced by that of oil. In our opinion, a
substantial CO2 regulatory consideration and incentive
via, for example, appropriately priced CO2 certificates,
would catalyze the development and implementation of
many technologies. That the material integration of such
different markets as fine chemicals, food, and plastics
becomes a reality seems obviously beneficial, but the
question is when it will be done. We are very optimistic
about the many roles biotechnological conversion of non-
conventional feedstocks will play in the circular economy,
not least to help the chemical industry to switch to
sustainable resource utilization and re-utilization.
Conflict of interest statement
Nothing declared.
Acknowledgements
Lars M. Blank and Kevin O’Connor received funding from the European
Union’s Horizon 2020 research and innovation program under grant
agreement no. 633962 for the project P4SB. Lars M. Blank was supported
by the Biotechnology Research and Information Network AG (BRAIN AG)
and by the German Federal Ministry of Education and Research (BMBF) as
part of the Strategic Alliance Zero Carbon Footprint (Grant no. FKZ
031A217F). The laboratory of Lars M. Blank was also partially funded by
the Deutsche Forschungsgemeinschaft (DFG, German Research
Foundation) under Germany’s Excellence Strategy within the Clusters of
Excellence TMFB 236 and FSC 2186 ‘The Fuel Science Center’. He also
acknowledges the Ministry of Culture and Science within the framework of
the NRW Strategieprojekt BioSC (No. 313/323-400-002 13) in the project
PlastiCycle. Kevin O’Connor and Tanja Narancic are supported by Science
Foundation Ireland research centre Grant No. SFI/16/RC/3889.
Appendix A. Supplementary data
Supplementary material related to this article can be
found, in the online version, at doi:https://doi.org/10.
1016/j.copbio.2019.11.011.
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