Bioresource Technology 344 (2022) 126307

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Strategies for Biosynthesis of C1 Gas-derived Polyhydroxyalkanoates:

A review
Jihee Yoon , Min-Kyu Oh *
Department of Chemical and Biological Engineering, Korea University, Seongbuk-gu, Seoul 02841, Republic of Korea

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Bioplastics production using cheaper

carbon source is crucial for cost
reduction.
• Various strategies to improve PHA pro­
duction from C1 gases were
accomplished.
• Autotrophs, acetogens, and methano­
trophs were optimized and engineered
for PHA.
• Process optimization and PHA copol­
ymer production from C1 gases were
carried out.
• C1 refinery to bioplastics is prospective
methodology to ease environmental
issues.

A R T I C L E I N F O A B S T R A C T

Keywords: Biosynthesis of polyhydroxyalkanoates (PHAs) from C1 gases is highly desirable in solving problems such as
Polyhydroxyalkanoates climate change and microplastic pollution. PHAs are biopolymers synthesized in microbial cells and can be used
Bioplastic as alternatives to petroleum-based plastics because of their biodegradability. Because 50% of the cost of PHA
Carbon dioxide
production is due to organic carbon sources and salts, the utilization of costless C1 gases as carbon sources is
Carbon monoxide
Methane
expected to be a promising approach for PHA production. In this review, strategies for PHA production using C1
gases through fermentation and metabolic engineering are discussed. In particular, autotrophs, acetogens, and
methanotrophs are strains that can produce PHA from CO2, CO, and CH4. In addition, integrated bioprocesses for
the efficient utilization of C1 gases are introduced. Biorefinery processes from C1 gas into bioplastics are pro­
spective strategies with promising potential and feasibility to alleviate environmental issues.

1. Introduction
Petroleum-based plastics, including poly(ethylene terephthalate)
(PET), high-density poly(ethylene) (HDPE), low-density poly(ethylene)
(LDPE), poly(vinyl chloride) (PVC), poly(propylene) (PP), and poly
(styrene) (PS), have desirable physical properties that include lightness,
resistance, rigidity, and inexpensive manufacture. The convenience and
ease have spurred the increased demand for plastics. In 2019, the global
production of plastics reached 368 million tons (da Costa, 2021). Plastic
waste can be handled by recycling, incineration, and landfill deposition,

* Corresponding author.
E-mail address: mkoh@korea.ac.kr (M.-K. Oh).

https://doi.org/10.1016/j.biortech.2021.126307
Received 26 September 2021; Received in revised form 4 November 2021; Accepted 5 November 2021
Available online 9 November 2021
0960-8524/© 2021 Elsevier Ltd. All rights reserved.
J. Yoon and M.-K. Oh
or being leaked. Globally in 2016, 16% of plastic waste was collected for
recycling, 25% incinerated, 40% reclaimed, and 19% dumped or leaked
unmanageably (Hundertmark et al., 2018). While the incineration of
plastics can be utilized to produce energy, it emits harmful byproducts in
the form of carbon dioxide and toxic pollutants such as dioxins, furans,
polychlorinated biphenyls and hydrochloric acid (Meereboer et al.,
2020; Verma et al., 2016). In addition, only half of the incinerated
plastic can be converted into energy in South Korea (Jang et al., 2020).
Thus, much of the plastic waste was not recycled. Even worse, plastic
waste that is not disposed of properly can enter watercourses. One
consequence is the accumulation of garbage in the ocean, such as the
Great Pacific garbage patch (Lebreton et al., 2018). These threaten the
marine ecosystem and return to humans through the food chain in a
microplastic form (Auta et al., 2017). Microplastics can accumulate in
the human body, especially in the liver, as demonstrated by in vivo ex­
periments (Im et al., 2021; Shi et al., 2021). This has spurred research
concerning the use of bioplastics as alternatives to petroleum-based
plastics.
Bioplastics are plastics that are biologically-based and/or, biode­
gradable. Bioplastics having both characteristics include poly­
hydroxyalkanoates (PHAs), poly(lactic acid) (PLA), poly(butylene
succinate) (PBS), and starch-based biopolymers (Abioye et al., 2018).
PHA is regarded as a promising alternative to petroleum-based plastics
since it rapidly and completely degrades in terrestrial and marine en­
vironments. The market size of PHAs in 2020 was estimated to be $62
million USD and is projected to reach $121 million USD by 2025
(Market, 2019). Especially, it is noteworthy that PHA production using
greenhouse gases (GHGs) is being commercialized by Newlight Tech­
nologies (Huntington Beach, CA, USA).
PHA is polymerized in vivo using various hydroxyalkanoyl-CoAs.
More than 160 different monomers have been used for PHA polymeri­
zation. The composition of the monomers determines the polymer
properties. Thus, polymers with desired properties can be produced
(Bhatia et al., 2021; Choi et al., 2020). A major limitation to the use of
Fig. 1. Closed-loop bioplastic cycle from C1 gases to PHAs.
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Bioresource Technology 344 (2022) 126307
PHA is the manufacturing expense. Approximately 50% of the PHA
production cost is spent on raw materials, such as carbon sources, other
nutrients, and salts (Li & Wilkins, 2020). To reduce this cost, the use of
cheaper and discarded resources, such as lignocellulose, wastewater, or
C1 gases, has been explored (Favaro et al., 2019; Morgan-Sagastume
et al., 2014; Tamis et al., 2014; Vigneswari et al., 2021). In particular,
C1 gases can be obtained at essentially no cost from industrial and
agricultural sources, and from household waste. Furthermore, since C1
gases are the main causes of global warming, their use as microbial
feedstocks for PHA production could help alleviate the environmental
crises of global warming and plastic waste.
In this review, recent advances in PHA production strategies from
carbon dioxide (CO2), carbon monoxide (CO), and methane (CH4) are
discussed. The metabolic characteristics of autotrophs, acetogens, and
methanotrophs for the utilization of C1 gases are described. Strategies to
facilitate PHA production, such as fermentation or genetic engineering,
are discussed. These methodologies allow the establishment of a com­
plete closed-loop PHA production from C1 gases and its degradation to
C1 gases (Fig. 1).
2. Metabolic conversion of C1 gases to feedstocks
Fossil fuel-based industries have been developed, resulting in air
pollution and global warming. The global average temperature in 2020
has increased by 1.3℃ compared to that at pre-industrial temperatures
(Hansena et al., 2021). Various attempts have been made to alleviate
atmospheric environmental issues. These include CO2 capture, renew­
able energy development, transportation improvements, and biorefinery
of C1 gases. Biorefinery using C1 gases as microbial feedstocks for the
production of value-added compounds has been accomplished using
autotrophs, acetogens, and methanotrophs. To utilize C1 gases as carbon
sources by those strains, stirred-tank reactor (STR) has been commonly
used with mixed gases (CO2/CO/CH4/H2/N2) injected into the culture
broth. Because C1 gases have very low solubility to water, additional
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

Fig. 2. Metabolic pathways for C1 gas-utilizing microorganisms. (A) Metabolism of autotrophic bacteria, such as cyanobacteria and the non-photoautotrophic
bacterium, C. necator, which utilize the Calvin-Benson-Bassham (CBB) cycle. (B) Metabolism of autotrophic bacteria, such as A. woodii, which utilize the Wood-
Ljungdahl pathway (WLP). This pathway also fixes CO, as denoted in red letters. (C) Metabolism of type II methanotrophic bacteria, which utilize serine cycle
and EMCP for methane assimilation. Abbreviations; PS (photosystem), 3PGA (3-phosphoglyceric acid), 1,3-BPG (1,3-bisphosphoglyceric acid), G3P (glyceraldehyde
3-phosphate), F1,6P (fructose 1,6-bisphosphate), F6P (fructose 6-phosphate), Ru5P (ribulose 5-phosphate), RuBP (ribulose 1,5-diphosphate), PEP (phosphoenol­
pyruvate), Pyr (pyruvate), PHB (poly-3-hydroxybutyrate), OAA (oxaloacetate), EMCP (ethylmalonyl-CoA pathway).

3
J. Yoon and M.-K. Oh
Fig. 2. (continued).
process modifications have been attempted, such as high rotational
speed of impeller, microbubble sparger, or increased partial pressure of
gases for enhancing gas–liquid mass transfer (Bae et al., 2021). Addi­
tional energy sources other than C1 gases, such as light and/or H2 are
provided to the reactor when autotrophs or acetogens are cultured. This
section mainly discusses the metabolic features of each microorganism
when utilizing C1 gas.
2.1. CO2 as a microbial feedstock
CO2 is a major contributor to global warming, and the amount of
emission is steadily increasing. Several microbial metabolic pathways
can fix CO2. These include the Calvin-Benson-Bassham (CBB) cycle,
Wood-Ljungdahl pathway (WLP), reductive tricarboxylic acid (TCA)
cycle, reductive glycine pathway, dicarboxylate/4-hydroxybutyrate
cycle, 3-hydroxypropionate-4-hydroxybutyrate cycle, and 3-hydroxy­
propionate cycle (Jiang et al., 2021). Since PHA-producing autotrophs
usually utilize the CBB cycle or WLP, these pathways are the focus in this
section.
Owing to the stable molecular structure of CO2, autotrophs that
utilize CO2 as a carbon source receive energy from light (photoauto­
trophs) or other compounds, such as H2 or formate (lithoautotrophs)
(Duerre & Eikmanns, 2015; Gleizer et al., 2019; Lau et al., 2015). Cya­
nobacteria are typical photoautotrophic bacteria that utilize CBB cycle
(Fig. 2A). These bacteria fix CO2 through the carboxylation reaction
with ribulose 1,5-biosphosphate (RuBP) catalyzed by ribulose 1,5-
bisphosphate carboxylase/oxygenase (RuBisCO), forming 3-phospho­
glycerate (3-PGA). 3-PGA can be converted to phosphoenolpyruvate
(PEP), which enters the central metabolism and is converted to biomass,
and also be converted to glyceraldehyde 3-phosphate (G3P), which
drives the CBB cycle (Humphreys & Minton, 2018; Sarma et al., 2016).
When light is provided, the energy for the CBB cycle is supplemented by
the oxidation of water catalyzed by photosystem II. Electrons pass
through the electron transport chain composed of the cytochrome
complex and photosystems and generate NADPH (Lai & Lan, 2015;
Vermaas, 2001). Inorganic compounds, such as H2, can also be utilized
as an energy source. A typical CBB cycle-metabolizing non-photoauto­
trophic bacterium is Cupriavidus necator (formerly Ralstonia eutropha).
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Bioresource Technology 344 (2022) 126307
C. necator obtains energy for CO2 fixation by oxidizing H2 to two H+
atoms and two electrons catalyzed by several membrane-bound [Ni-Fe]
hydrogenases (Burgdorf et al., 2005; Schäfer et al., 2013). Electrons are
transported through [Fe-S] clusters in the respiratory chain, and NADPH
or NADH is produced (Panich et al., 2021). Both cyanobacteria and
C. necator naturally produce PHA.
Acetogens are strictly anaerobic bacteria that produce acetate as the
major product when grown on CO2 through the WLP. Clostridium sp. and
Acetobacterium sp. belong to acetogen (Vidra & Németh, 2018). WLP is
the most energy efficient pathway for CO2 fixation (Jiang et al., 2021)
(Fig. 2B). The WLP consists of a methyl branch and a carbonyl branch. In
the methyl branch, CO2 is reduced to formate by formate dehydrogenase
(FDH). In turn, formate is converted to methyl-corrinoid iron-sulfur
protein (methyl-CoFeSP) through a series of reactions involving tetra­
hydrofolate (THF). In the carbonyl branch, CO2 is converted to CO in a
reaction catalyzed by CO dehydrogenase/acetyl-CoA synthase (CODH/
Acs). The reactions in the two branches convert methyl-CoFeSP and CO
are to acetyl-CoA as catalyzed by the bifunctional enzyme CODH/Acs.
Membrane-bound Rnf complex and flavin-based electron bifurcation
provide NAD(P)H and reduced ferredoxin (Fd2-) as reducing power by
oxidizing H2 when fixing CO2 (Schoelmerich & Müller, 2019; Schuch­
mann & Müller, 2014). Instead of hydrogen, electrotrophic metabolism
has been verified by supplying electrons through the electrode (Lovley,
2011).
2.2. CO as a microbial feedstock
An enormous amount of CO is released from industrial processes,
such as thermal power generation and steel manufacture. The CO can be
combusted to CO2 for energy generation. However, the efficiency of this
process is low due to inert gases. Also, CO can increase the risk of ex­
plosion (Choi et al., 2017). Thus, the utilization of CO as a microbial
feedstock is a sustainable and environmentally-friendly strategy. Recent
studies have examined CO utilization for production of biochemicals
(Fernández-Naveira et al., 2016; Moreira et al., 2021; Wilkins & Atiyeh,
2011). Utilization of CO can damage biological functions that include
oxygen transport and is toxic to most organisms (Ernst & Zibrak, 1998).
Only some bacteria, such as acetogens, can utilize CO as a carbon source.
J. Yoon and M.-K. Oh
As mentioned above, acetogen metabolism of CO is accomplished
through the WLP. While CO can be utilized directly in the carbonyl
branch, CO is oxidized to CO2 by CODH in the methyl branch, providing
reducing power (Fig. 2B). Rhodospirillum rubrum is a typical producer of
PHA from CO (Do et al., 2007). Also, it is accessible to convert CO into
soluble substrates, such as acetate or formate, through acetogens for
synthesis of PHA (Fei et al., 2021; Hwang et al., 2020).
2.3. CH4 as a microbial feedstock
CH4 is a GHG with a global warming potential 25 times greater than
that of CO2 (Ford et al., 2012). More than 60% of emitted CH4 is from
anthropogenic sources that include animal husbandry, wastewater
treatment, natural gas refining, and composting (La et al., 2018). In
addition, natural gas is composed of CH4, which makes it easy to secure
(Liu et al., 2020). Utilization of CH4 as a microbial feedstock is an
optimistic strategy because of its’ pronounced reduction potential,
which is higher than that of glucose, serving improvement of product
yield (Lee et al., 2016). Utilization of CH4 as microbial feedstock is
available both anaerobically and aerobically by methanotrophs (Duerre
& Eikmanns, 2015). Mostly, in anerobic conditions, enrichment cultures
are utilized, while pure cultures are used in aerobic conditions (Gambelli
et al., 2018; Zhu et al., 2012).
Methanotrophs utilize CH4 by oxidizing it to methanol as catalyzed
by methane monooxygenase (MMO). A soluble form (sMMO) and a
particulate form (pMMO) existed (Fig. 2C). Methanol is then converted
into formaldehyde by periplasmic pyrroloquinoline quinone (PQQ)-
dependent methanol dehydrogenase. Formaldehyde can be assimilated
through two pathways that divide methanotrophs into two groups. Type
I methanotrophs belonging to γ-proteobacteria utilize the RuMP
pathway. Type II methanotrophs belonging to α-proteobacteria have a
serine cycle and ethylmalonyl-CoA pathway (EMCP) for formaldehyde
assimilation (Jiang et al., 2021). Because type II methanotrophs natu­
rally produce PHA, many studies have focused on the type II microor­
ganisms in the production of PHA from methane. In type II
methanotrophs, formaldehyde is passed through the assimilative or
dissimilative pathways. It is first oxidized to formate and finally into CO2
by FDH, providing NADH for energy. Formaldehyde can also be con­
verted into methylene-H4F, which enters the serine cycle in the assim­
ilative pathway. Serine is then converted into PEP in a series of
sequential reactions and is used for biomass production. Glyoxylate is
required for the continuous operation of the serine cycle. To regenerate
glyoxylate, type II methanotrophs use the EMCP anaplerotic reaction
that involves various CoA esters. Methanotrophs can even produce poly
(3-hydroxybutyrate) (PHB) through EMCP intermediates, acetoacetyl-
CoA, and 3-hydroxybutyryl-CoA.
3. Strategies for production of PHAs from C1 gases
3.1. Biosynthetic pathway of PHAs
PHAs are biodegradable polyesters synthesized inside cells using
various hydroxyalkanoyl-CoAs as monomers. The monomers determine
properties of PHAs, such as stiffness, tensile strength, and melting
temperature. PHAs are polymerized via the PHA synthetic pathway
starting with acetyl-CoA as catalyzed by 3-ketothiolase, acetoacetyl-CoA
reductase, and PHA synthase (Fig. 2). The major physiological role of
PHAs is storage of carbon and energy when carbon is in excess and when
other nutrients, such as nitrogen or phosphorus, are limited (Tan et al.,
2014). When nutrients are abundant, acetyl-CoA is used to operate the
TCA cycle to obtain energy and grow. Coenzyme A (CoA) produced by
the TCA cycle interferes with PHA synthesis. Conversely, when nutrients
are lacking, the low CoA level does not interfere with PHA synthesis, and
acetyl-CoA is converted to PHA. The best-known PHA producer is
C. necator under both autotrophic and heterotrophic conditions
(Vigneswari et al., 2021). The phaCAB operon in C. necator is involved in
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Bioresource Technology 344 (2022) 126307
PHA synthesis. The operon harbors genes that encode PHA synthase,
acetoacetyl-CoA reductase, and 3-ketothiolase.
An important advantage of PHAs is that various monomers can be
copolymerized with 3-hydroxybutyrate (3HB, C4) with the shortest R
group in its basic structure. Depending on the number of carbon atoms in
the copolymerized monomer, it comprises short chain length (SCL; C3-
C5) and medium chain length (MCL; C6-C14) PHAs. PHB is the most
well-known and commercially available SCL-PHA. Two acetyl-CoA
molecules condense to acetoacetyl-CoA, which is involved in 3-ketothio­
lase encoded by phaA. Acetoacetyl-CoA is then hydrated to 3-hydroxy­
butyryl-CoA by NADPH-dependent acetoacetyl-CoA reductase encoded
by phaB. 3-Hydroxybutyryl-CoA is polymerized as catalyzed by PHA
synthase encoded by phaC. The phaCAB operon was initially demon­
strated in C. necator (Choi et al., 2020; Schubert et al., 1988). PHB is
arranged in an (R)-configuration with a right-handed double helix,
resulting in a high degree of crystallinity, stiffness, and melting tem­
perature (Anjum et al., 2016; D’Amico et al., 2012; Marlina et al., 2018;
Reichardt & Rieger, 2011). To improve the properties of PHB, copoly­
merization or blends have been attempted. The typical SCL-PHA
copolymer is poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV).
To produce PHBV, precursors that include propionate or valerate are
usually supplemented to support 3-hydroxyvalerate (3HV) (Berezina &
Yada, 2016; Matsumoto et al., 2011). Otherwise, some wild type bac­
teria including archaea can produce PHBV without any feeding pre­
cursors (Koller, 2019). Genetic engineering for PHBV has also been
performed in the absence of precursors (Orita et al., 2014; Yoon et al.,
2021). For example, Methylorubrum extorquens is a model strain of
methylotroph that is capable of serine cycle and EMCP metabolism using
propionyl-CoA as a major metabolite. To utilize propionyl-CoA, which is
a precursor of 3HV, for copolymerization, the essential gene phaA for
EMCP was deleted and the alternative gene bktB from C. necator was
introduced (Yoon et al., 2021).
MCL-PHAs are typically produced from Pseudomonas sp. The poly­
merization of MCL-PHAs is dependent on PHA synthases (Ray & Kalia,
2017). There are four groups of PHA synthases (Class I–IV) based on
their primary structure, substrate specificity, and subunit composition.
Class I has a homodimer structure with a PhaC subunit and is specific to
SCL-hydroxyalkanoyl-CoAs (Sagong et al., 2018). C. necator has a class I
PHA synthase and mainly produces PHB and copolyesters with 3HV and
4-hydroxybutyrate (4HB). Class II PHA synthase also has a homodimer
structure with the PhaC1 or PhaC2 subunits. It has broad substrate
specificity and is capable of polymerizing MCL-hydroxyalkanoyl-CoAs.
P. aeruginosa, P. putida, P. resinovorans, and some other bacteria have
Class II PHA synthases. When volatile fatty acids are supplemented, they
produce MCL-PHAs, such as poly(3-hydroxybutyrate-co-3-hydroxyhex­
anoate) (PHBHHx), or other PHAs that include 3-hydroxyheptanoate
(3HHp), 3-hydroxyoctanoate (3HO), 3-hydroxynonanoate (3HN), or 3-
hydroxydecanoate (3HD) (Cerrone et al., 2014; Mezzolla et al., 2018).
Class III and IV are heterodimers consisting of PhaC/PhaE and PhaC/
PhaR, respectively. Both are specific for SCL-hydroxyalkanoyl-CoAs
(Pradani et al., 2020). To produce MCL-PHAs, recombinant strains
were constructed by introducing a Class II PHA synthase. For example,
phaC for Class I PHA synthase of C. necator was replaced by phaC1 and
phaC2 from Rhodococcus aetherivorans. Recombinant C. necator can
produce PHBHHx from plant oil. Furthermore, phaJ involved in
β-oxidation was overexpressed, resulting in increased 3-hydroxyhexa­
noate (3HHx) portion in PHA (Budde et al., 2011).
3.2. Strategies for PHA synthesis from CO2
3.2.1. Photoautotrophic synthesis of PHB
Diverse cyanobacteria that grow photoautotrophically have been
tested for their PHB-producing ability using CO2 and light. Synecho­
coccus sp., Nostoc muscorum, Aulosira fertilissima, Scytonema sp., and
others produce PHB within 10% per dry cell weight (DCW) (Bhati et al.,
2010; Tarawat et al., 2020) (Table 1). To improve PHB production,
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

Table 1
Strategies of PHB production from CO2.
Strain Strategies Cultivation Cell PHA PHA PHA Reference
period mass titer content productivity

Photoautotrophic bacteria in autotrophic condition

Anacystis nidulans Cultivation of diverse cyanobacteria under 7 days n.d. n.d. 4.4% (w/ n.d. (Bhati et al., 2010)
photoautotrophy w)
Aulosira fertilissima 14 days n.d. n.d. 6.5% (w/ n.d.
w)
Calothrix sp. 28 days n.d. n.d. 6.8% (w/ n.d.
w)
Nostoc muscorum 21 days n.d. n.d. 8.5% (w/ n.d.
w)
Csytonema sp. 21 days n.d. n.d. 7.4% (w/ n.d.
w)
Tolypothrix distorta TISTR Cultivation of diverse cyanobacteria under 20 days n.d. n.d. n.d. 2.2 mg/ (Tarawat et al.,
8985 photoautotrophy (gDCW∙day) 2020)
Nostoc muscorum TISTR 20 days n.d. n.d. n.d. 0.8 mg/
8871 (gDCW∙day)
Synechocystis sp. PCC Introduction of heterologous phaABEC genes from 8 days 0.153 0.0106 7% (w/ 0.0000552 g/ (Hondo et al., 2015)
6803GT P. aeruginosa and cultivation with nitrogen g/L g/L w) (L∙h)
limitation
Synechocystis sp. PCC Deletion of pta and ach and introduction of xfpK 40 days ~ 2 g/L 0.232 12% (w/ 0.000304 g/ (Carpine et al., 2017)
6803 from Bifidobacterium breve g/L w) (L∙h)
Synechocystis sp. PCC Random mutation using UV lightCultivation 500 h 2.11 g/ 0.78 g/L 37% (w/ 0.00154 g/ (Kamravamanesh
6714 without N and P L w) (L∙h) et al., 2018)
Synechocystis sp. PCC Knock-out of exoD 14 days 2.06 g/ ~ 0.33 ~ 16% n.d. (Mittermair et al.,
6714 mutant Mt_a24 L g/L (w/w) 2021)
S. elongatus UTEX2973 Introduction of heterologous phaCAB genes from 10 days 1.22 g/ 0.278 21.1% 0.00116 g/ (Roh et al., 2021)
C. necator and cultivation with nitrogen limitation L g/L (w/w) (L∙h)
Mixed wastewater-borne Cultivation depending on light cycle and nutrient 8 days n.d. 0.104 5.7% (w/ 0.000542 g/ (Arias et al., 2018)
cyanobacteria deficiency (N/P) g/L w) (L∙h)
Mixed culture of Separation of Bio-module I (CO2 → sucrose) and 16 days n.d. 0.156 n.d. 0.000992 g/ (Löwe et al., 2017)
Synechococcus Bio-module II (sucrose → PHA) g/L (L∙h)
elongatus cscB and
Pseudomonas putida
cscAB
Mixed culture of Separation of Bio-module I (CO2 → sucrose) and Observation n.d. n.d. 31% (w/ 0.00118 g/ (Weiss et al., 2017)
Synechococcus Bio-module II (sucrose → PHB) for 5 months w) (L∙h)
elongatus PCC 7942
and Halomonas
boliviensis
Photoautotrophic bacteria in mixotrophic condition
Synechocystis sp. PCC Deletion of pirC (regulatory protein of intracellular 28 days n.d. n.d. 81% (w/ n.d. (Koch et al., 2020)
6803 glycogen and PHB pool) and introduction of phaAB w)
from C. necatorCultivation in CO2 + light with
supplementation of acetate
Synechocystis sp. PCC Deficiency of P with supplement of acetate and 10 days n.d. n.d. 38% (w/ n.d. (Panda & Mallick,
6803 fructose w) 2007)
Synechocystis sp. PCC Adjustment of nutrient and light 12 days n.d. n.d. 21.8% n.d. (Monshupanee &
6803 intensityCultivation with deficiency of N and P and (w/w) Incharoensakdi,
supplement of glucose 2014)
Scytonema geitleri Adjustment of pH, temperature and carbon sources 28 days n.d. n.d. 7.12% n.d. (Singh et al., 2019)
Bharadwaja such as sucrose and acetate (w/w)
Non-photoautotrophic bacteria in autotrophic condition
C. necator Development of a bench-plant scale, recycled-gas, 40 h 91.3 g/ 61.9 g/L 67.8% 1.55 g/(L∙h) (Tanaka et al., 1995)
closed-circuit culture system by maintaining L (w/w)
oxygen concentration in the gas phase below the
limit for a gas explosion (6.9%)
C. necator Cultivation in air-lift fermenter by addition of 96 h 69.3 g/ 56.4 g/L 81.4% 1.02 g/(L∙h) (Taga et al., 1997)
sodium carboxymethylcellulose to culture medium L (w/w)
C. necator Set up of mathematical model based on mass 120 h 60 g/L 49.2 g/L 82% (w/ 0.41 g/(L∙h) (Mozumder et al.,
balance and cultivation in continuous stirred-tank w) 2015)
reactor (CSTR) with an air-lift fermenter
C. necator H16 Cultivation using low content of H2 and nitrogen 144 h 0.39 g/ 0.27 g/L 70% (w/ 0.00188 g/ (Miyahara et al.,
limitation L w) (L∙h) 2020)
C. eutrophus B-10646 Chemostat batch culture using third stage, which 80 h 45 ± 5 36 g/L 80 ± 5% 0.45 g/(L∙h) (Volova et al., 2013)
stages are depending on amount of nitrogen supply g/L (w/w)
C. necator DSM545 Two-stage cultivation system composed of phase About 63 h 21 g/L 16 g/L 74% (w/ 0.252 g/(L∙h) (Garcia-Gonzalez
for heterotrophic cell mass growth and phase for w) et al., 2015)
autotrophic PHB production below explosion limit
of oxygen concentration 5%(vol)
Ideonella sp. O-1 Gas component: H2:O2:CO2 = 7:1:1Cultivation in 24 h 6.75 g/ 5.26 g/L 77.9% 0.219 g/(L∙h) (Tanaka et al., 2011)
minimal medium containing 1.0 g/L (NH4)2SO4 L (w/w)
P. denitrificans High-cell density fermentation with nitrogen About 132 h About About 4 57.3% About 0.0304 (Tanaka et al., 2016)
NBRC13301 limitation 7 g/L g/L (w/w) g/(L∙h)

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J. Yoon and M.-K. Oh
environmental conditions of pH and temperature were adjusted (Singh
et al., 2019), as were the intensity and cycle of lighting (Arias et al.,
2018; Monshupanee & Incharoensakdi, 2014). In addition, because PHB
tends to accumulate when nutrients, such as nitrogen (N) and phos­
phorus (P), are limited, two-stage cultivation methods have often been
used. The first stage promoted cell growth with sufficient supplemen­
tation of N and P. The second stage induced PHB production in nutrient-
deficient medium. (Arias et al., 2018; Kamravamanesh et al., 2018;
Miyahara et al., 2020; Roh et al., 2021; Tanaka et al., 2016; Volova et al.,
2013).
To improve photoautotrophic PHB production, the use of mixotrophs
or mixed cultures have also been attempted. A mixotrophic culture in­
volves cultivation with other heterotrophic substrates and autotrophic
substrates. This strategy can provide sufficient energy and carbon,
leading to improved cell growth and PHB production. Compared to
glucose, acetate, and fructose as heterotrophic substrates, acetate is a
good nutrient for increase of PHB production (Koch et al., 2020; Mon­
shupanee & Incharoensakdi, 2014; Panda & Mallick, 2007; Singh et al.,
2019). Acetate may be easily converted to acetyl-CoA, which is the
precursor of PHB. A mixed culture of cyanobacteria with other bacteria
is another strategy for PHB production from CO2 (Arias et al., 2018). The
cyanobacterium Synechococcus elongatus exports sucrose using fixed CO2
with a high efficiency of up to 85%. Co-culture of heterotrophic bacteria
for PHB production using exported sucrose can increase PHB produc­
tivity. For example, in mixed cultures S. elongatus cscB produced sucrose
from CO2 photoautotrophically and P. putida cscAB produced PHA from
sucrose. By applying this system, 156 mg/L of PHB was produced (Löwe
et al., 2017). Likewise, the co-culture of S. elongatus PCC 7942 and
Halomonas boliviensis enabled PHB production from CO2 by separating
the reaction of CO2 with sucrose and that of sucrose to PHB (Weiss et al.,
2017).
Genetic engineering has been used to improve PHB productivity in
cyanobacteria. A typical way to improve PHB production is to introduce
heterologous PHB synthesis pathways to strengthen PHB synthesis
fluxes. Thus, phaCAB and phaABEC from C. necator and Microcystis aer­
uginosa NIES-843, respectively, were introduced into Synechococcus sp.,
resulting in an increase in PHB content (PHB per DCW (w/w)) by 12-fold
compared to the control (Hondo et al., 2015; Roh et al., 2021). Another
example is the enhancement of the acetyl-CoA pool in Synechocystis sp.
PCC 6803, given that acetyl-CoA is a precursor of PHB. For this, phos­
photransacetylase encoded by pta and acetyl-CoA hydrolase encoded by
ach were knocked out, and xfpK from Bifidobacterium breve was intro­
duced (Carpine et al., 2017). Furthermore, the regulatory protein of PHB
synthesis was engineered in Synechocystis sp. PCC 6803. PirC is a regu­
lator involved in the conversion of intracellular glycogen and PHB pools
under nutrient-limiting conditions. When pirC was deleted and phaAB
from C. necator was introduced, the PHB content per DCW reached 81%
with acetate supplement (Koch et al., 2020). In cyanobacteria, over­
expression of a high copy number is achieved via plasmid or integration.
However, the approach is not cost-effective in large-scale reactors
because it requires the addition of antibiotics. To solve this problem, an
antibiotic-free stable expression plasmid system was developed by
constructing a cyanobacterial lethal recA null mutant and recA-
harboring plasmid. Using this system, PHB accumulation was improved
by stably expressing phaCAB (Akiyama et al., 2011).
Random mutations can be an effective strategy to improve produc­
tivity. Random mutations in Synechocystis sp. PCC 6714 using ultraviolet
light resulted in PHB accumulation of 37% per DCW (Kamravamanesh
et al., 2018). In this mutant, further rational engineering with the
deletion of exoD, which is involved in exopolysaccharide (EPS) pro­
duction, reduced EPS production and increased PHB production (Mit­
termair et al., 2021). Techno-economic analysis for PHB production
using CO2 has not yet been reported. However, the synthesis of cyano­
bacterial PHB is expected to reduce cost by up to approximately 50%
when compared to the synthesis of heterotrophic bacterial PHB (Costa
et al., 2018).
7
Bioresource Technology 344 (2022) 126307
3.2.2. Non-photoautotrophic synthesis of PHB from CO2
Several studies have addressed PHB production from CO2 using non-
photoautotrophs that use inorganic compounds, such as H2, as an energy
source. C. necator is a typical chemolithotroph and PHA producer. PHA
production by C. necator in both autotrophic and heterotrophic condi­
tions has been studied (Cavalheiro et al., 2009; Dalsasso et al., 2019).
Since C. necator also tends to accumulate PHB when mineral elements,
such as N or P, are limited, two-stage cultivation methods and further
three-stage cultivation methods that control the supply of N or P were
suggested to induce PHB production using CO2. For example, a three-
stage chemostat batch culture was set up for C. necator. In the first
stage, the amount of N for sufficient growth was 100 mg/g cells. The
PHB content per DCW was 8% at that stage. In the second stage, nitrogen
supplementation was limited to 60 mg/g cells. The content of accumu­
lated PHB was 47%. Finally, in the third stage, N was not supplied. The
PHB content reached 80% (Volova et al., 2013). A mathematical model
based on mass balance was applied to predict PHB production under
various conditions, such as oxygen, nitrogen stress, and gas composition
(O2/H2) (Mozumder et al., 2015). H2 is an energy source for CO2 fixation
by oxidization catalyzed by membrane-bound [Ni-Fe] hydrogenases.
However, it has a risk of explosion. To reduce the risk, a low H2 content
of 3.6% was adopted to produce PHB in C. neactor (Miyahara et al.,
2020). Similar efforts were made to lower the risk of explosion when
culturing using O2, another combustible element, by cultivating
C. necator with a lowered oxygen concentration in gaseous substrates
below 6.9% (Garcia-Gonzalez et al., 2015; Tanaka et al., 1995).
Chemolithotrophs other than C. necator have also been used for PHA
production. Paracoccus denitrificans reportedly produced 57.3% PHB per
DCW from CO2 and H2 under N-limited conditions (Tanaka et al., 2016).
Ideonella sp. O-1 also produced 77.9% PHB per DCW from CO2 and H2,
and displayed higher tolerance to CO compared to that of C. necator,
leading to the expectation that industrial exhaust gas containing CO
would be utilized directly (Tanaka et al., 2011). The strategy of con­
verting CO2 into PHB using hydrogen-oxidizing bacteria is one of the
most promising and feasible production platforms, given that it achieved
high concentration of PHB with competitive productivity (Table 1).
3.2.3. Synthesis of PHA copolymers using CO2
PHA copolymers have various properties depending on the monomer
composition. Several strategies, such as feeding precursors or genetic
engineering, have been explored for the production of PHA copolymers
using CO2. Table 2 lists native producers of PHA copolymers under
autotrophic conditions without any supplement of precursors (Table 2).
Oscillatoria okeni can produce MCL-PHA PHBHHx with 3HHx 5.5%(mol)
in PHA under N-limited conditions (Taepucharoen et al., 2017) and
Anabaena spiroides TISTR 8075 produces PHBV (Tarawat et al., 2020). In
addition, PHA copolymers can be produced by adding precursors
(Table 2). γ-Butyrolactone, valerate, and hexanoate are utilized to pro­
duce 4HB, 3HV, and 3HHx, respectively, in C. necator (Park et al., 2014;
Volova et al., 2013).
In the absence of supplementation of precursors, metabolic engi­
neering can enable the synthesis of PHA copolymers. In C. necator,
heterologous expression of phaCs from Pseudomonas sp. and supple­
mentation with acrylic acid, which inhibits β-oxidation, caused the
production of MCL-PHAs with C4-C6 monomers (Nangle et al., 2020). In
addition, overexpression of phaAB and bktB and introduction of phaC1
from Pseudomonas sp. 61–3 yielded high levels of PHA per DCW with
monomers, 3HV 1.2%(mol) and 3-hydroxy-4-methylvalerate (3H4MV)
1.2%(mol) in autotrophic conditions (Miyahara et al., 2020).
3.3. PHA synthesis from CO
3.3.1. Native CO-utilizing bacteria
Synthetic gas (syngas) is produced through the thermochemical
process of gasification. In this process, the materials are not completely
oxidized and create gases that contain a large amount of CO (Drzyzga
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

Table 2
Strategies of PHA copolymers from CO2.
Strain Strategies for PHA copolymer production Substrates PHA composition Reference

Oscillatoria okeni Cultivation with nitrogen limitation CO2 + light PHBV (3HV 5.5% [mol]) (Taepucharoen et al.,
TISTR 8549 2017)
Anabaena spiroides Cultivation with various carbon sources CO2 + light PHBV (3HV 43.2% [mol]) (Tarawat et al.,
TISTR 8075 CO2 + light + acetate PHBV (3HV 24.9% [mol]) 2020)
CO2 + light + PHBV (3HV 17.6% [mol])
propionate
CO2 + light + valerate PHBV (3HV 51.1% [mol])
Supplement of precursors
C. necator ATCC Supplement of valeric acid CO2 + H2 + valerate PHBV (3HV 46% [mol]) (Park et al., 2014)
17,697
C. necator DSM 545 Mixotrophic cultivation and supplement of valeric acid Glucose + CO2 + H2 + PHBV (3HV 50% [mol]) (Ghysels et al., 2018)
valerate
C. eutrophus B-10646 Autotrophic Supplement of PHA precursors such as valerate, CO2 + H2 + valerate PHBV (3HV 85.1% [mol]) (Volova et al., 2013)
hexanoate, and γ-butyrolactone after cell growth CO2 + H2 + hexanoate PHBHHx (3HHx 20.0% [mol])
CO2 + H2 + P(3HB-co-4HB) (4HB 51.3% [mol])
γ-butyrolactone
Metabolic engineering
C. necator H 16 Expression of heterologous phaCs and the addition of acrylic CO2 + H2 MCL-PHAs (4 ≤ chain length ≤ 14) (Nangle et al., 2020)
acid (β-oxidation inhibitor)
C. necator H 16 Introduction of phaC1 from Pseudomonas sp. 61–3 and CO2 + H2 PHA terpolymer with 3HV 1.2% (Miyahara et al.,
overexpression of phaAB and bktB and 3H4MV 1.2% [mol] 2020)

et al., 2015). Gas composition is usually 40% CO, 40% H2, 10% CO2, and grown on CO, CO dehydrogenase and CO-insensitive hydrogenase
10% N2 (Karmann et al., 2019). R. rubrum produces PHB from CO as a convert CO to CO2 and H2. Although R. rubrum harbors a CBB cycle, 13C
carbon and energy source under anaerobic conditions (Table 3). When analysis and gene expression analysis have demonstrated that

Table 3
Strategies for PHA production from CO.
Strain Strategies Cultivation Cell mass PHA titer PHA content PHA Reference
period productivity

Native CO-utilizing bacteria

Rhodospirillum rubrum Fed-batch cultivation with three About 180 h 5.32 g/L PHB 1.6 g/ PHB 30.0% (w/w) About PHB (Karmann
different growth phases under nutrient L 0.0089 g/(L∙h) et al., 2019)
(C, P)-limited conditions with
supplement of acetate
Rhodospirillum rubrum Optimization of culture method with 5 days 0.45 g/L PHB 0.036 PHB 8% (w/w) PHB 0.0003 g/ (Mongili &
supplement of acetate and fructose g/L (L∙h) Fino, 2021)
Rhodospirillum rubrum Cultivation with supplement of acetate 5 days n.d. n.d. PHB 28% (w/w) n.d. (Revelles
and analysis of central carbon et al., 2016)
metabolites using 13C-labelling acetate
in co-substrates condition
Rhodospirillum rubrum Optimization of gasifier and supplement n.d. 0.00708 n.d. PHBV 38% (w/w) with PHBV 0.00247 (Do et al.,
of acetate g/(L∙h) monomer composition of g/(L∙h) 2007)
86%(mol) 3HB and 14%
(mol) 3HV
Rhodospirillum rubrum Deletion of endogenous phaC1 and 5 days 1 g/L P (3HD-co- P (3HD-co-3HO) 6.7% P (3HD-co-3HO) (Heinrich
phaC2 and introduction of phaC1, phaG, 3HO) (w/w) with monomer 0.000558 g/ et al., 2016)
and gene number PP_0763 encoding 0.067 g/L composition of 42.8% (L∙h)
CoA ligase from P. putida (mol) 3HO and 57.0%
(mol) 3HD
Seliberia Optimization of CO concentration and 56 h 20.2 g/L 12.3 g/L PHB 61–63% (w/w) 0.22 g/(L∙h) (Volova
carboxydohydrogena limitation of N and sulfur et al., 2015)
Z-1062
Clostridium Construction of PHB-producing 24 h 0.482 g/L PHB PHB 5.61% (w/w) PHB 0.00113 g/ (Lemgruber
autoethanogenum recombinatn by introducing phaCAB 0.02704 g/ (L∙h) et al., 2019)
from C. necator and comparison of PHB L
content depending on H2 supply
Clostridium coskatii Construction of PHB-producing 48 h OD 0.43 n.d. PHB 1.12% (w/w) n.d. (Flüchter
recombinant by introducing thlA, hbd, et al., 2019)
crt, phaJ and phaEC from Clostridium
species
Non-native CO-utilizing bacteria
C. necator Introduction of cox subcluster (genes for 21 days 2.62 g/L PHB 1.30 PHB 49.7% (w/w) PHB 0.00258 g/ (Heinrich
carbon monoxide dehydrogenase and g/L (L∙h) et al., 2018)
the maturation of CODH) from
Oligotropha carboxidovorans
C. necator In vitro conversion of CO into CO2 72 h 33.8 g/L PHB 14.2 PHB 42% (w/w) 0.197 g/(L∙h) (Shin et al.,
through nanoscaled cellulose particles g/L 2021)
with enzyme complex (carbon
monoxide dehydrogenase, carbon
monoxide binding unit, and carbonic
anhydrase)

8
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

carboxylases other than RuBisCO incorporate CO2 from syngas to

biomass (Revelles et al., 2017; Revelles et al., 2016). Acetate supple­

(Lagoa-Costa
et al., 2017)

et al., 2020)

et al., 2018)
(Al Rowaihi
Reference

(Fei et al.,
mentation can permit sufficient production of PHB from CO using

(Hwang
R. rubrum (Do et al., 2007; Mongili & Fino, 2021). Limiting nutrients,

2021)
such as P, can lead to a PHB content per DCW of up to 30% (Karmann
et al., 2019). Another study engineered R. rubrum to produce MCL-PHA

6.5% (w/w)PHB 1.57 g/L with

PHB 0.5 g/L with content per

3HB 1.02 g/L with yield 0.26
copolymers from CO. To incorporate MCL monomers, endogenous

content 46.6% (w/w) when

PHB content per DCW 24%

PHB 0.1 g/L with content

phaC1 and phaC2 were knocked out, phaG (3-hydroxyacyl carrier pro­

using purchased formate

tein thioesterase), phaC1 (MCL-PHA synthase), and gene number

DCW 33.3% (w/w)

PP_0763 (MCL-fatty acid coenzyme A ligase) from P. putida were
expressed under the control of a CO-inducible promoter. This recombi­
nant R. rubrum accumulated up to 7.1% PHA per DCW with a monomer

Product
composition of 42.8%(mol) 3HO and 57.0%(mol) 3HD (Heinrich et al.,

(w/w)

g/g
2016). Seliberia carboxydohydrogena Z-1062 also produces PHB from CO.
Adjustment of the CO concentration and limiting N and sulfur reportedly
led to a PHB content of 62.8% with a productivity rate of 0.22 g/(L∙h)

Cultivation using acetate solution from Process I

Fed-batch cultivation inducing consumption of

expressing phaAB from C. necator and pct from

acetate and ethanol in N limitation condition
(Volova et al., 2015).

Clostridium beijerinckiiCultivation in complex

productionFed-batch cultivation in minimal

with supplement of fructose 30 g/L for cell

Introduction of 3HB synthesis pathway by
Engineering of Clostridium sp. to produce PHB is an alternative

Overexpression of ftfL and phaCAB for

improvements of cell growth and PHB
strategy to utilize CO (Table 3). Clostridium species can grow on CO
through the WLP. However, the bacteria are non-native producers of
PHB. Therefore, some Clostridium species with PHB production capacity

medium with yeast extract

were constructed by heterologous expression of the PHB synthesis
pathway from C. necator and Pseudomonas sp. (Flüchter et al., 2019;
Lemgruber et al., 2019). In particular, metabolomics, transcriptomics,
and genome-scale metabolic models have shown that cellular redox
states, such as ATP availability, limit PHB production, rather than the

Strategies

medium
acetyl-CoA pool, in C. autoethanogenum capable of producing PHB pro­

harvest
duction (Lemgruber et al., 2019).
A techno-economic cost analysis was conducted to determine
whether the production of PHB by R. rubrum using syngas is economi­

Engineered E. coli
cally viable. The estimated cost was $1.65/kg (Choi et al., 2010), which

Methylorubrum
Enriched PHA
accumulating
is economically feasible. Further research on the utilization of syngas is

consortium

Engineered
Process II

extorquens

C. necator
anticipated.
Strain

3.3.2. Synthetic CO-utilizing bacteria

Synthetic CO-utilizing C. necator was developed for PHB production
from syngas (Table 3). CO oxidation was facilitated by expressing the

Formate 53.1 mM
LEthanol 2.22 g/
Acetate 4.976 g/

cox subcluster consisting of CODH and accessory proteins for CODH

L2,3-butanediol

Acetate 3.2 g/L

Acetate 4 g/L

maturation from Oligotropha carboxidovorans. This allowed the conver­

1.632 g/L

sion of CO to CO2 and growth in the presence of syngas. Engineered

Product

C. necator produced up to 49.7% PHB per DCW (Heinrich et al., 2018). In

another study, nanoscale cellulose particles which were attached with
CODH, carbon monoxide binding unit, and carbonic anhydrase were
conditions such as pH, CO concentrations and

Cultivation of gas mixture of CO2:H2 = 15:85

(acetate production) and solventogenic phase
Consisting of 2 phases with acidogenic phase

immobilized on the surface of C. necator. The complex catalyzed the

under elevated pressure for H2 solubility
(ethanol and 2,3-butanediol production)

conversion of CO to CO2 in vitro. C. necator utilizing CO2 as carbon

biocatalystOptimization of catalytic

source produced 14.2 g/L PHB with PHB content of 42% per DCW (Shin
Conversion process with whole-cell

et al., 2021). The findings indicated the potential for PHB production
Combined processes for Syngas into PHAs through intermediates

from syngas.
Cultivation using syngas

3.3.3. Process development of syngas for PHA production: Combining two

bioconversion processes
Among syngas components, CO is toxic and the production of PHA
Strategies

from CO is usually poor. Processes combining the two bioconversion

buffer

processes have been developed to overcome these limitations. Inte­

grated bioconversion processes consisting of Process I (CO → organic
substrate) and Process II (organic substrate → PHAs) are effective in
terms of CO fixation, H2 consumption, substrate cost, and safety (Garcia-
autoethanogenum

Gonzalez & De Wever, 2018) (Table 4). For conversion of syngas in

Acetobacterium

Process I, Clostridium species were cultivated to produce acetate, which

Clostridium

Clostridium

biocatalyst
Whole cell
liungdahlii
Process I

A. woodii

was used to synthesize PHB or monomer 3HB through a PHB-

Strain

woodii

accumulating consortium or engineered Escherichia coli, respectively,

in Process II (Fei et al., 2021; Lagoa-Costa et al., 2017). Furthermore,
combined processes using formate as an intermediate were developed.
Formate
Syngas →

Syngas →

Syngas →
Acetate

Acetate

Acetate
→ PHB

→ PHB

→ PHB
→ 3HB

In Process I, syngas was converted into formate using a whole-cell bio­

product
Table 4

CO2 →
Final

catalyst, and in Process II, engineered M. extorquens produced PHB

(Hwang et al., 2020). Similarly, two bioconversion processes were

9
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

applied to convert CO2 to PHB through acetate (Table 4). A. woodii and
C. necator were cultivated to convert CO2 into acetate and convert ac­
etate into PHB (Al Rowaihi et al., 2018).
3.4. PHA synthesis from CH4
3.4.1. Enrichment culture to utilize waste CH4 and produce PHB
Enrichment cultures provide diverse metabolic activities and make
particular microorganisms to favor growth in certain surroundings. Such
cultures are usually used for biorefining of waste CH4 emitted from
landfills and in anaerobic digestion processes (Table 5). Methanotrophic
consortia from landfill biomass, compost soils, or anaerobic digester
sludge reportedly produced PHB from waste CH4, with the accumulation
of 47.8–51% (w/w) PHB per DCW (Chidambarampadmavathy et al.,
2015; Fergala et al., 2018). While most methanotrophs prefer low
temperatures (<30 ◦ C), thermophilic methanotrophs produce PHBs at
55–58 ◦ C. For example, cultivation of methanotrophic consortia domi­
nated by thermophilic methanotroph Methylcaldum szegediense OR2
resulted PHB production (Luangthongkam et al., 2019a). When using
methanotrophic enrichment culture, a two-stage cultivation method
with growth phase (sufficient nutrients) and PHB production phase
(nutrient limitation) was also used to induce PHB accumulation in cells
(Helm et al., 2008). In addition to the limited supply of N, P, sulfur, or
iron, diverse factors, including CH4/O2 ratio, CH4 mass transfer,

Table 5
Strategies for PHB production from CH4.
Strain Strategies Cultivation Cell mass PHB PHB PHB Reference
period titer content productivity

Enriched culture
Methylocystis sp.- Two-stage cultivation method separating 48 h n.d. n.d. 33.6% n.d. (Helm et al., 2008))
dominated growth phase and PHB accumulation (w/w)
methanotrophic phase with potassium deficiency
enrichment
Methylocystis-dominated Influence of temperature during culture 19 days Specific n.d. 35.1% n.d. (Pérez et al., 2019)
methanotrophic enrichment and cultivation with nitrogen growth (w/w)
enrichment limitation rate 0.05
h− 1
Methylocystis, Methyldopa, Effect of CH4/O2 ratios on PHB n.d. About 0.8 About 8.5% n.d. (Karthikeyan et al., 2015)
and production g/L 0.068 g/ (w/w)
Pseudoxanthomonas- L
dominated consortium
Methylocystis-dominated Demonstration of stability for 29 months 29 months n.d. n.d. 46.2% 1.13 g/(L∙h) (Helm et al., 2006)
enriched culture in an open system with methane (w/w)
Methylophilus, New setup for PHB production by n.d. n.d. n.d. 59.4% n.d. (Salem et al., 2021)
Methylocystis- recycling the biomass after PHB phase (w/w)
dominated enriched
culture
Methanotrophic Comparison of PHB capacity between 10 days OD 0.316 n.d. 47.88% n.d. (Chidambarampadmavathy
consortium from two methanotrophic consortium (w/w) et al., 2015)
landfill top-cover and
compost soils
Methanotrophs Development of accelerating CH4 mass 7 days OD 0.7 n.d. PHB n.d. (Myung et al., 2016a)
transfer without agitation (emulsion- 32% (w/
based fermentation) w)
Methanotrophic Production from anaerobic digester 70 days Specific n.d. 51% 0.0095 g/ (Fergala et al., 2018)
enrichment culture sludge growth (L∙h)
rate 0.078
h− 1
Methanotrophic- Optimization of Cu concentration and 72 h n.d. n.d. 48.7% n.d. (Zhang et al., 2018)
heterotrophic culture CH4:O2 ratio and cultures derived from (w/w)
seed sludge after about 320 days of
enrichment
Thermophilic Optimization of temperature (55℃ or n.d. Specific n.d. 2.4% n.d. (Luangthongkam et al.,
methanotrophic 58℃) and supplement of acetate growth (w/w) 2019a)
enrichment rate
1
0.0104 h−
Pure culture
Methylosinus trichosporium Optimization of medium, air inlet, and 120 h 18 g/L 9 g/L 50% (w/ 0.075 g/(L∙h) (Shah et al., 1996)
OB3b agitation speed w)
Methylosinus trichosporium Optimization of carbon source (methane n.d. 0.0928 g/L 0.0487 52.5% n.d. (Zaldívar Carrillo et al.,
OB3b or methanol), nitrogen source g/L (w/w) 2018)
(ammonium or nitrate), and nitrogen-to-
carbon ratioCultivation in mixotrophic
condition with methanol
Methylocystis sp. GB 25 Separation of growth phase and PHB 24 h n.d. n.d. 51% n.d. (Wendlandt et al., 2001)
DSMZ 7674 production phase
Methylocystis hirsuta Effect of magnesium and 144 h 8.0 g/L 5.87 g/L 73.4% 0.0408 g/ (Ghoddosi et al., 2019)
phosphorusCultivation in mixotrophic (L∙h)
condition with methanol and ethanol
Methylocystis hirsuta CSC1 Effect of O2:CH4 ratio, temperature, and 16 days n.d. n.d. 45% (w/ n.d. (Rodríguez et al., 2020a)
nitrogen source w)
Methylocystis hirsuta Application in a novel gas-recycling About 68 About 5 g/ n.d. 34.6% n.d. (García-Pérez et al., 2018)
bubble column bioreactor with limitation days L (w/w)
of potassium, manganese, and nitrogen
Methylocystis hirsuta Optimal operation of bubble column 65 days 3.0 g/L n.d. 14.5% n.d. (Rodríguez et al., 2020b)
bioreactor (w/w)

10
J. Yoon and M.-K. Oh Bioresource Technology 344 (2022) 126307

temperature, and Cu concentration, have been optimized (Karthikeyan
et al., 2015; Myung et al., 2016b; Pérez et al., 2019; Zhang et al., 2018).
In particular, novel methods for PHB production have been developed.
For example, water-in-oil emulsions were used to stimulate CH4 mass
transfer without agitation, given that the solubility of CH4 is higher in oil
than in water (Myung et al., 2016b). Further, a new setup for meth­
anotrophic enrichment culture for higher PHB production was estab­
lished by recycling the biomass after the PHB phase. In this system, the
PHB content reached 59.4% DCW (Salem et al., 2021). This methano­
trophic enrichment was operated stably in an open system for 29
months, with repeated multiple continuous growth and polymer for­
mation processes. The productivity of PHB from CH4 was 1.13 g/(L∙h)
(Helm et al., 2006).
A techno-economic evaluation estimated the cost of PHB production
from methane as $4.1–6.8/kg. Furthermore, thermophilic methano­
trophs could reduce costs by $3.2–5.4/kg, because refrigerants were not
required. Compared to sugar-based PHB production processes, with an
estimated cost of $7.5/kg, the estimated costs were lower. This is
because the cost of raw material was estimated to be only 22% when
using CH4, while it was approximately 50% when using sugars (Levett
et al., 2016).
3.4.2. Pure culture of methanotrophs for PHB production
Pure cultures of type II methanotrophs can be used because they are
native PHB producers via the serine cycle and EMCP (Table 5). Similar to
other PHB production strategies, two-stage cultivation methods sepa­
rating the growth phase and PHB production phase have been applied to
methanotrophs (Wendlandt et al., 2001). Optimization of the medium,
air supply, agitation, carbon source, nitrogen ratio, O2:CH4 ratio, and
temperature was accomplished for cell growth and PHB production in
Methylocystis hirsuta or Methylosinus trichosporium (Ghoddosi et al.,
2019; Rodríguez et al., 2020a; Shah et al., 1996; Zaldívar Carrillo et al.,
2018). In addition, mixotrophic conditions by supplementing with
methanol or ethanol were applied to increase PHB production (Ghoddosi

Table 6
Strategies for PHA copolymers production from CH4.
Strain Strategies for PHA copolymer production Substrates PHA content and its Reference
composition

Supplement of precursors
Methylosinus-dominated Supplement of propionic acid and valeric acid CH4 + Propionic PHBV 3.5% (w/w) (3HV (Luangthongkam
methanotrophic acid 22.6% (mol)) et al., 2019b)
enrichment CH4 + Valeric acid PHBV 14.1% (w/w) (3HV
65.0% (mol))
Methanotrophic Supplement of valeric acid CH4 + Valeric acid PHBV 52% (w/w) (3HV 33% (Fergala et al., 2018)
enrichment culture (mol))
Thermophilic Supplement of propionic acid and valeric acid CH4 + Propionic PHBV about 3% (w/w) (3HV (Luangthongkam
methanotrophic acid 85% (mol)) et al., 2019a)
enrichment CH4 + Valeric acid PHBV about 10% (w/w) (3HV
75%(mol))
Methylocystis parvus OBBP Supplement of propionic acid and valeric acid CH4 PHB 50% (w/w) (Myung et al.,
CH4 + Propionic PHBV 32% (w/w) (3HV 8% 2016a)
acid (mol))
CH4 + Valeric acid PHBV 54% (w/w) (3HV 22%
(mol))
Methylocystis parvus OBBP Supplement of ω-hydroxyalkanoate monomers CH4 PHB 42% (w/w) (Myung et al., 2017)
CH4 + Butyrate PHB 55% (w/w)
CH4 + 3- PHB 59% (w/w)
hydroxybutyrate
CH4 + 4- P(3HB-co-4HB) 50% (w/w)
hydroxybutyrate (4HB 9.5% (mol))
CH4 + Valerate PHBV 54% (w/w) (3HV 25%
(mol))
CH4 + 5- P(3HB-co-3HV-co-5HV) 48%
hydroxyvalerate (w/w) (3HV 1.4% (mol) and
5HV 3.6% (mol))
CH4 + Hexanoate PHB 56% (w/w)
CH4 + 6- P(3HB-co-4HB-co-6HHx) 48%
hydoxyhexanoate (w/w) (4HB 1% (mol) and
6HHx 1.4% (mol))
Methylocystis sp. WRRC1 Supplement of valerate or n-pentanol CH4 PHB 30% (w/w) (Cal et al., 2016)
CH4 + Valerate PHBV 60% (w/w) (3HV 50%
(mol))
Methylocystis hirsuta Supplement of versatile fatty acids Biogas PHB 43.1% (w/w) (López et al., 2018)
Biogas + Acetic acid PHB 52.3% (w/w)
Biogas + Propionic PHBV 47.9% (w/w) (3HV 2%
acid (mol))
Biogas + Butyric PHB 52.2% (w/w)
acid
Biogas + Valeric PHBV 53.8% (w/w) (3HV 25%
acid (mol))
Methylobacterium Supplement of citrate or propionate CH4 + Citrate P(3HB-co-3HV-co-3HO) (3HV (Zuñiga et al., 2013)
organophilum CZ-2 35% (mol) and 3HO 10%
(mol))
CH4 + Propionate PHBV (3HV 25% (mol))
Metabolic engineering
Methylosinus trichosporium Introduction of 4HB synthesis pathway by expressing sucD (CoA- CH4 P(3HB-co-4HB) 7.01% (w/w) (Nguyen & Lee,
OB3b dependent succinate semialdehyde dehydrogenase), 4hbD (4- (4HB 3.08% (mol)) 2021)
hydroxybutyrate dehydrogenase) from Porphyromonas gingivalis
or yqhD (NADPH-dependent succinate semialdehyde reductase)
from E. coliTwo-stage cultivation without N deficiency

11
J. Yoon and M.-K. Oh
et al., 2019; Zaldívar Carrillo et al., 2018). Furthermore, for M. hirsuta, a
mechanistic model was applied to determine the optimal O2:CH4 ratio
(Chen et al., 2020), and a novel gas-recycling bubble column reactor was
developed and optimized (García-Pérez et al., 2018; Rodríguez et al.,
2020b).
3.4.3. Synthesis of PHA copolymers from CH4
The simplest way to produce PHA copolymers is to add related fatty
acids, such as propionate or valerate (Table 6). PHBV was synthesized
when propionate or valerate was added to both enrichment cultures and
pure cultures (Cal et al., 2016; Fergala et al., 2018; López et al., 2018;
Luangthongkam et al., 2019a; Luangthongkam et al., 2019b; Myung
et al., 2016a; Nguyen & Lee, 2021). Supplementation of 5-hydroxyvaler­
ate (5HV) and 6-hydroxyhexanoate (6HHx) resulted in the synthesis of
PHA terpolymers P(3HB-co-3HV-co-5HV) and P(3HB-co-4HB-co-6HHx),
respectively, in Methylocystis parvus OBBP (Myung et al., 2017). Further
supplementation with citrate resulted in P(3HB-co-3HV-co-3HO) in
Methylobacterium organophilum (Zuñiga et al., 2013).
Recently, metabolic engineering of M. trichosporium OB3b success­
fully synthesized P(3HB-co-4HB) using CH4 as the sole carbon source.
For the intracellular synthesis of 4HB, sucD encoding CoA-dependent
succinate semialdehyde dehydrogenase and 4hbD encoding 4-hydroxy­
butyrate dehydrogenase from Porphyromonas gingivalis were intro­
duced. Furthermore, to increase the succinyl-CoA pool, isocitrate
dehydrogenase encoded by icd was overexpressed, resulting in the
synthesis of P(3HB-co-4HB) with a 4HB portion of 3.08% (mol) using
CH4 (Nguyen & Lee, 2021).
4. Future perspectives
C1 gases are the main culprits of global warming. Recently, to ach­
ieve carbon neutrality, various bioproducts involving biopolymers,
biochemicals, and biofuels have been produced from C1 gases. Espe­
cially, PHAs, biodegradable polyesters, are promising products due to
their complete biodegradability in compost and marine conditions, of­
fering the way to solve the problem of petroleum-based plastics. Many
strategies for utilizing CO2, CO, and CH4 have been carried out to
improve PHA production. Along with optimization of culture conditions,
metabolic engineering for upregulation of PHA synthesis pathway
contributed to the increase in PHA productivity. Also, it facilitated
synthesis of various PHA copolymers such as PHBV, P(3HB-co-4HB),
PHBHHx, and MCL-PHAs. However, there are still some drawbacks. i)
PHA content per DCW is usually low. When growing on C1 gases,
assimilative fluxes are accelerated, which means that acetyl-CoA, a
precursor of PHA, is mainly used for growth rather than production of
PHA. Given that mixotrophic culture with acetate supplement serves for
improvement of PHA content per DCW, genetic engineering for increase
of intracellular acetyl-CoA pool can serve the solution. Adaptive labo­
ratory evolution or random mutation might be another way to improve
PHA content, as high-throughput screening using fluorescent dye as
BODIPY or Nile red is available. ii) Although growth has been improved
through many studies on culture optimization, there is still a limit to
high cell density. This may be improved through mixotrophic cultiva­
tion with balance of carbon sources. Further, as gene manipulation
techniques and synthetic biology tools have been improved, strain
development is being performed more efficiently and systematically.
Therefore, engineering to improve the assimilative carbon flux from C1
gases and increase PHA production would be accelerated. iii) It is
necessary to produce various PHA copolymers because the physical
properties of PHB, such as stiffness and high melting point, are not
proper as commercial plastics. So far, most studies depend on supple­
ment of precursors when producing PHA copolymers, which is not cost-
effective. With the developed genetic engineering technologies, such as
CRISPR-Cas system, it potentializes the production of non-natural
polyesters, which contain monomers such as phenyllactate, glycolate,
or 2-hydroxyisovalerate, as well as SCL-, and MCL-PHAs. Studies for
12
Bioresource Technology 344 (2022) 126307
PHA copolymer production having better physical and chemical prop­
erties through metabolic engineering would be one of the important
points for commercializing PHA copolymer produced from C1 gases.
Overall, PHA production using C1 gas is influential and significant in
terms of providing the closed-loop bioplastic system, where produced
bioplastics are degraded to produce new bioplastics. When PHAs are
processed through aerobic biodegradation or incinerated after use, CO2
or CO is generated. When the bioplastics are anaerobically digested, CH4
is produced. These gases can be used to produce PHAs using the strains
reviewed in this manuscript. These recycling system of PHAs can be
sustainable and environmentally benign. It is certain that improvement
of PHA productivity and diversification of PHA species using C1 gas
would make it a competitive platform.
5. Conclusion
PHAs are attractive alternatives to conventional plastics since they
are biodegradable. However, manufacturing costs are high. Reducing
these costs by the use of inexpensive raw materials is desirable. C1 gases,
such as CO2, CO, and CH4, are cheap or cost-free substrates, as they can
be obtained from industrial or agricultural processes, landfills, and
household waste. This review introduces the strategies for PHA pro­
duction using C1 gases, including cultivation optimization, metabolic
engineering, and development of novel processes. As PHA production
processes using GHGs are being commercialized, this review will inform
the recent progress of the PHA production platforms.
CRediT authorship contribution statement
Jihee Yoon: Conceptualization, Formal analysis, Investigation, Data
curation, Writing – original draft, Writing – review & editing. Min-Kyu
Oh: Conceptualization, Writing – original draft, Writing – review &
editing, Supervision, Project administration, Funding acquisition.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
This research was supported by C1 Gas Refinery Program through
the National Research Foundation of Korea (NRF) funded by the Min­
istry of Science and ICT (2015M3D3A1A01064919).
References
Abioye, O.P., Abioye, A.A., Afolalu, S.A., Ongbali, S.O., 2018. A review of biodegradable
plastics in Nigeria. International Journal of Mechanical Engineering and Technology
(IJMET) 9 (10).
Akiyama, H., Okuhata, H., Onizuka, T., Kanai, S., Hirano, M., Tanaka, S., Sasaki, K.,
Miyasaka, H., 2011. Antibiotics-free stable polyhydroxyalkanoate (PHA) production
from carbon dioxide by recombinant cyanobacteria. Bioresource technology 102
(23), 11039–11042.
Al Rowaihi, I.S., Kick, B., Grötzinger, S.W., Burger, C., Karan, R., Weuster-Botz, D.,
Eppinger, J., Arold, S.T., 2018. A two-stage biological gas to liquid transfer process
to convert carbon dioxide into bioplastic. Bioresource Technology Reports 1, 61–68.
Anjum, A., Zuber, M., Zia, K.M., Noreen, A., Anjum, M.N., Tabasum, S., 2016. Microbial
production of polyhydroxyalkanoates (PHAs) and its copolymers: a review of recent
advancements. International journal of biological macromolecules 89, 161–174.
Arias, D.M., Uggetti, E., García-Galán, M.J., García, J., 2018. Production of
polyhydroxybutyrates and carbohydrates in a mixed cyanobacterial culture: effect of
nutrients limitation and photoperiods. New biotechnology 42, 1–11.
Auta, H.S., Emenike, C., Fauziah, S., 2017. Distribution and importance of microplastics
in the marine environment: a review of the sources, fate, effects, and potential
solutions. Environment international 102, 165–176.
Bae, J., Song, Y., Lee, H., Shin, J., Jin, S., Kang, S., Cho, B.-K., 2021. Valorization of C1
gases to value-added chemicals using acetogenic biocatalysts. Chemical Engineering
Journal 131325.
J. Yoon and M.-K. Oh
Berezina, N., Yada, B., 2016. Improvement of the poly (3-hydroxybutyrate-co-3-
hydroxyvalerate) (PHBV) production by dual feeding with levulinic acid and sodium
propionate in Cupriavidus necator. New biotechnology 33 (1), 231–236.
Bhati, R., Samantaray, S., Sharma, L., Mallick, N., 2010. Poly-β-hydroxybutyrate
accumulation in cyanobacteria under photoautotrophy. Biotechnology journal 5
(11), 1181–1185.
Bhatia, S.K., Otari, S.V., Jeon, J.-M., Gurav, R., Choi, Y.-K., Bhatia, R.K., Pugazhendhi, A.,
Kumar, V., Banu, J.R., Yoon, J.-J., 2021. Biowaste-to-bioplastic
(polyhydroxyalkanoates): Conversion technologies, strategies, challenges, and
perspective. Bioresource Technology 124733.
Budde, C.F., Riedel, S.L., Willis, L.B., Rha, C., Sinskey, A.J., 2011. Production of poly (3-
hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil by engineered Ralstonia
eutropha strains. Applied and environmental microbiology 77 (9), 2847–2854.
Burgdorf, T., Lenz, O., Buhrke, T., Van Der Linden, E., Jones, A.K., Albracht, S.P.,
Friedrich, B., 2005. [NiFe]-hydrogenases of Ralstonia eutropha H16: modular
enzymes for oxygen-tolerant biological hydrogen oxidation. Journal of molecular
microbiology and biotechnology 10 (2–4), 181–196.
Cal, A.J., Sikkema, W.D., Ponce, M.I., Franqui-Villanueva, D., Riiff, T.J., Orts, W.J.,
Pieja, A.J., Lee, C.C., 2016. Methanotrophic production of polyhydroxybutyrate-co-
hydroxyvalerate with high hydroxyvalerate content. International journal of
biological macromolecules 87, 302–307.
Carpine, R., Du, W., Olivieri, G., Pollio, A., Hellingwerf, K.J., Marzocchella, A., dos
Santos, F.B., 2017. Genetic engineering of Synechocystis sp. PCC6803 for poly-
β-hydroxybutyrate overproduction. Algal research 25, 117–127.
Cavalheiro, J.M., de Almeida, M.C.M., Grandfils, C., Da Fonseca, M., 2009. Poly (3-
hydroxybutyrate) production by Cupriavidus necator using waste glycerol. Process
biochemistry 44 (5), 509–515.
Cerrone, F., Choudhari, S.K., Davis, R., Cysneiros, D., O’Flaherty, V., Duane, G.,
Casey, E., Guzik, M.W., Kenny, S.T., Babu, R.P., 2014. Medium chain length
polyhydroxyalkanoate (mcl-PHA) production from volatile fatty acids derived from
the anaerobic digestion of grass. Applied microbiology and biotechnology 98 (2),
611–620.
Chen, X., Rodríguez, Y., López, J.C., Muñoz, R., Ni, B.-J., Sin, G.r. 2020. Modeling of
Polyhydroxyalkanoate Synthesis from Biogas by Methylocystis hirsuta. ACS
Sustainable Chemistry & Engineering, 8(9), 3906-3912.
Chidambarampadmavathy, K., Karthikeyan, O.P., Heimann, K., 2015. Biopolymers made
from methane in bioreactors. Engineering in Life Sciences 15 (7), 689–699.
Choi, D., Chipman, D.C., Bents, S.C., Brown, R.C., 2010. A techno-economic analysis of
polyhydroxyalkanoate and hydrogen production from syngas fermentation of
gasified biomass. Applied biochemistry and biotechnology 160 (4), 1032–1046.
Choi, E.S., Min, K., Kim, G.-J., Kwon, I., Kim, Y.H., 2017. Expression and characterization
of Pantoea CO dehydrogenase to utilize CO-containing industrial waste gas for
expanding the versatility of CO dehydrogenase. Scientific reports 7, 44323.
Choi, So Young, Rhie, Mi Na, Kim, Hee Taek, Joo, Jeong Chan, Cho, In Jin, Son, Jina,
Jo, Seo Young, Sohn, Yu Jung, Baritugo, Kei-Anne, Pyo, Jiwon, Lee, Youngjoon,
Lee, Sang Yup, Park, Si Jae, 2020. Metabolic engineering for the synthesis of
polyesters: A 100-year journey from polyhydroxyalkanoates to non-natural
microbial polyesters. Metabolic engineering 58, 47–81.
Costa, J.A.V., Moreira, J.B., Lucas, B.F., Braga, V.d.S., Cassuriaga, A.P.A., Morais, M.G.d.
2018. Recent advances and future perspectives of PHB production by cyanobacteria.
Industrial Biotechnology, 14(5), 249-256.
D’Amico, D.A., Manfredi, L.B., Cyras, V.P., 2012. Relationship between thermal
properties, morphology, and crystallinity of nanocomposites based on
polyhydroxybutyrate. Journal of Applied Polymer Science 123 (1), 200–208.
da Costa, J.P. 2021. The 2019 Global pandemic and plastic pollution prevention
measures: playing catch-up. Science of The Total Environment, 145806.
Dalsasso, R.R., Pavan, F.A., Bordignon, S.E., de Aragão, G.M.F., Poletto, P., 2019.
Polyhydroxybutyrate (PHB) production by Cupriavidus necator from sugarcane
vinasse and molasses as mixed substrate. Process Biochemistry 85, 12–18.
Do, Y.S., Smeenk, J., Broer, K.M., Kisting, C.J., Brown, R., Heindel, T.J., Bobik, T.A.,
DiSpirito, A.A., 2007. Growth of Rhodospirillum rubrum on synthesis gas: conversion
of CO to H2 and poly-β-hydroxyalkanoate. Biotechnology and Bioengineering 97 (2),
279–286.
Drzyzga, O., Revelles, O., Durante-Rodríguez, G., Díaz, E., García, J.L., Prieto, A., 2015.
New challenges for syngas fermentation: towards production of biopolymers.
Journal of Chemical Technology & Biotechnology 90 (10), 1735–1751.
Duerre, P., Eikmanns, B.J., 2015. C1-carbon sources for chemical and fuel production by
microbial gas fermentation. Current opinion in biotechnology 35, 63–72.
Ernst, A., Zibrak, J.D., 1998. Carbon monoxide poisoning. New England journal of
medicine 339 (22), 1603–1608.
Favaro, L., Basaglia, M., Casella, S., 2019. Improving polyhydroxyalkanoate production
from inexpensive carbon sources by genetic approaches: a review. Biofuels,
Bioproducts and Biorefining 13 (1), 208–227.
Fei, P., Luo, Y., Lai, N., Wu, H., 2021. Biosynthesis of (R)-3-hydroxybutyric acid from
syngas-derived acetate in engineered Escherichia coli. Bioresource Technology
125323.
Fergala, A., AlSayed, A., Khattab, S., Ramirez, M., Eldyasti, A., 2018. Development of
methane-utilizing mixed cultures for the production of polyhydroxyalkanoates
(PHAs) from anaerobic digester sludge. Environmental science & technology 52
(21), 12376–12387.
Fernández-Naveira, Á., Abubackar, H.N., Veiga, M.C., Kennes, C., 2016. Efficient
butanol-ethanol (BE) production from carbon monoxide fermentation by Clostridium
carboxidivorans. Applied microbiology and biotechnology 100 (7), 3361–3370.
Flüchter, S., Follonier, S., Schiel-Bengelsdorf, B., Bengelsdorf, F.R., Zinn, M., Dürre, P.,
2019. Anaerobic production of poly (3-hydroxybutyrate) and its precursor 3-
13
Bioresource Technology 344 (2022) 126307
hydroxybutyrate from synthesis gas by autotrophic clostridia. Biomacromolecules 20
(9), 3271–3282.
Ford, H., Garbutt, A., Jones, L., Jones, D.L., 2012. Methane, carbon dioxide and nitrous
oxide fluxes from a temperate salt marsh: Grazing management does not alter Global
Warming Potential. Estuarine, Coastal and Shelf Science 113, 182–191.
Gambelli, L., Guerrero-Cruz, S., Mesman, R.J., Cremers, G., Jetten, M.S., Op den
Camp, H.J., Kartal, B., Lueke, C., van Niftrik, L., 2018. Community composition and
ultrastructure of a nitrate-dependent anaerobic methane-oxidizing enrichment
culture. Applied and environmental microbiology 84 (3), e02186–17.
Garcia-Gonzalez, L., De Wever, H., 2018. Acetic acid as an indirect sink of CO2 for the
synthesis of polyhydroxyalkanoates (PHA): Comparison with PHA production
processes directly using CO2 as feedstock. Applied Sciences 8 (9), 1416.
Garcia-Gonzalez, L., Mozumder, M.S.I., Dubreuil, M., Volcke, E.I., De Wever, H., 2015.
Sustainable autotrophic production of polyhydroxybutyrate (PHB) from CO2 using a
two-stage cultivation system. Catalysis Today 257, 237–245.
García-Pérez, T., López, J.C., Passos, F., Lebrero, R., Revah, S., Muñoz, R., 2018.
Simultaneous methane abatement and PHB production by Methylocystis hirsuta in a
novel gas-recycling bubble column bioreactor. Chemical Engineering Journal 334,
691–697.
Ghoddosi, F., Golzar, H., Yazdian, F., Khosravi-Darani, K., Vasheghani-Farahani, E.,
2019. Effect of carbon sources for PHB production in bubble column bioreactor:
Emphasis on improvement of methane uptake. Journal Of Environmental Chemical
Engineering 7 (2), 102978.
Ghysels, S., Mozumder, M.S.I., De Wever, H., Volcke, E.I., Garcia-Gonzalez, L., 2018.
Targeted poly (3-hydroxybutyrate-co-3-hydroxyvalerate) bioplastic production from
carbon dioxide. Bioresource technology 249, 858–868.
Gleizer, S., Ben-Nissan, R., Bar-On, Y.M., Antonovsky, N., Noor, E., Zohar, Y., Jona, G.,
Krieger, E., Shamshoum, M., Bar-Even, A., 2019. Conversion of Escherichia coli to
generate all biomass carbon from CO2. Cell 179 (6).
Hansena, J., Satoa, M., Ruedyb, R., Schmidtc, G., Lob, K. 2021. Global Temperature in
2020. columbia.edu.
Heinrich, D., Raberg, M., Fricke, P., Kenny, S.T., Morales-Gamez, L., Babu, R.P.,
O’Connor, K.E., Steinbüchel, A., 2016. Synthesis gas (Syngas)-derived medium-
chain-length polyhydroxyalkanoate synthesis in engineered Rhodospirillum rubrum.
Applied and environmental microbiology 82 (20), 6132–6140.
Heinrich, D., Raberg, M., Steinbüchel, A., 2018. Studies on the aerobic utilization of
synthesis gas (syngas) by wild type and recombinant strains of Ralstonia eutropha
H16. Microbial biotechnology 11 (4), 647–656.
Helm, J., Wendlandt, K.D., Jechorek, M., Stottmeister, U., 2008. Potassium deficiency
results in accumulation of ultra-high molecular weight poly-β-hydroxybutyrate in a
methane-utilizing mixed culture. Journal of applied microbiology 105 (4),
1054–1061.
Helm, J., Wendlandt, K.D., Rogge, G., Kappelmeyer, U., 2006. Characterizing a stable
methane-utilizing mixed culture used in the synthesis of a high-quality biopolymer
in an open system. Journal of applied microbiology 101 (2), 387–395.
Hondo, S., Takahashi, M., Osanai, T., Matsuda, M., Hasunuma, T., Tazuke, A.,
Nakahira, Y., Chohnan, S., Hasegawa, M., Asayama, M., 2015. Genetic engineering
and metabolite profiling for overproduction of polyhydroxybutyrate in
cyanobacteria. Journal of bioscience and bioengineering 120 (5), 510–517.
Humphreys, C.M., Minton, N.P., 2018. Advances in metabolic engineering in the
microbial production of fuels and chemicals from C1 gas. Current opinion in
biotechnology 50, 174–181.
Hundertmark, T., Mayer, M., McNally, C., Simons, T.J., Witte, C., 2018. How plastics
waste recycling could transform the chemical industry. McKinsey & Company 12.
Hwang, H.W., Yoon, J., Min, K., Kim, M.-S., Kim, S.-J., Cho, D.H., Susila, H., Na, J.-G.,
Oh, M.-K., Kim, Y.H., 2020. Two-stage bioconversion of carbon monoxide to
biopolymers via formate as an intermediate. Chemical Engineering Journal 389,
124394.
Im, C., Kim, H., Zaheer, J., Kim, J.Y., Lee, Y.J., Kang, C.M., Kim, J.S., 2021. PET tracing
of biodistribution for orally administered 64Cu-labeled polystyrene in mice. Journal
of Nuclear Medicine. https://doi.org/10.2967/jnumed.120.256982.
Jang, Y.-C., Lee, G., Kwon, Y., Lim, J.-H., Jeong, J.-H., 2020. Recycling and management
practices of plastic packaging waste towards a circular economy in South Korea.
Resources, Conservation and Recycling 158, 104798.
Jiang, W., Villamor, D.H., Peng, H., Chen, J., Liu, L., Haritos, V., Ledesma-Amaro, R.,
2021. Metabolic engineering strategies to enable microbial utilization of C1
feedstocks. Nature Chemical Biology 17 (8), 845–855.
Kamravamanesh, Donya, Kovacs, Tamas, Pflügl, Stefan, Druzhinina, Irina, Kroll, Paul,
Lackner, Maximilian, Herwig, Christoph, 2018. Increased poly-β-hydroxybutyrate
production from carbon dioxide in randomly mutated cells of cyanobacterial strain
Synechocystis sp. PCC 6714: Mutant generation and characterization. Bioresource
technology 266, 34–44.
Karmann, S., Panke, S., Zinn, M., 2019. Fed-batch cultivations of Rhodospirillum rubrum
under multiple nutrient-limited growth conditions on syngas as a novel option to
produce poly (3-hydroxybutyrate)(PHB). Frontiers in bioengineering and
biotechnology 7, 59.
Karthikeyan, O.P., Chidambarampadmavathy, K., Nadarajan, S., Lee, P.K., Heimann, K.,
2015. Effect of CH4/O2 ratio on fatty acid profile and polyhydroxybutyrate content
in a heterotrophic–methanotrophic consortium. Chemosphere 141, 235–242.
Koch, M., Bruckmoser, J., Scholl, J., Hauf, W., Rieger, B., Forchhammer, K., 2020.
Maximizing PHB content in Synechocystis sp. PCC 6803: a new metabolic engineering
strategy based on the regulator PirC. Microbial Cell Factories 19 (1), 1–12.
Koller, M., 2019. Polyhydroxyalkanoate biosynthesis at the edge of water activity-
Haloarchaea as biopolyester factories. Bioengineering 6 (2), 34.
La, H., Hettiaratchi, J.P.A., Achari, G., Dunfield, P.F., 2018. Biofiltration of methane.
Bioresource technology 268, 759–772.
J. Yoon and M.-K. Oh
Lagoa-Costa, B., Abubackar, H.N., Fernández-Romasanta, M., Kennes, C., Veiga, M.C.,
2017. Integrated bioconversion of syngas into bioethanol and biopolymers.
Bioresource technology 239, 244–249.
Lai, M.C., Lan, E.I., 2015. Advances in metabolic engineering of cyanobacteria for
photosynthetic biochemical production. Metabolites 5 (4), 636–658.
Lau, N.-S., Matsui, M., Abdullah, A.A.-A., 2015. Cyanobacteria: photoautotrophic
microbial factories for the sustainable synthesis of industrial products. BioMed
research international 2015, 754934.
Lebreton, L., Slat, B., Ferrari, F., Sainte-Rose, B., Aitken, J., Marthouse, R., Hajbane, S.,
Cunsolo, S., Schwarz, A., Levivier, A., 2018. Evidence that the Great Pacific Garbage
Patch is rapidly accumulating plastic. Scientific reports 8 (1), 1–15.
Lee, O.K., Hur, D.H., Nguyen, D.T.N., Lee, E.Y., 2016. Metabolic engineering of
methanotrophs and its application to production of chemicals and biofuels from
methane. Biofuels, Bioproducts and Biorefining 10 (6), 848–863.
Lemgruber, R.d.S.P., Valgepea, K., Tappel, R., Behrendorff, J.B., Palfreyman, R.W., Plan,
M., Hodson, M.P., Simpson, S.D., Nielsen, L.K., Köpke, M. 2019. Systems-level
engineering and characterisation of Clostridium autoethanogenum through
heterologous production of poly-3-hydroxybutyrate (PHB). Metabolic engineering, 53,
14-23.
Levett, I., Birkett, G., Davies, N., Bell, A., Langford, A., Laycock, B., Lant, P., Pratt, S.,
2016. Techno-economic assessment of poly-3-hydroxybutyrate (PHB) production
from methane—The case for thermophilic bioprocessing. Journal of Environmental
Chemical Engineering 4 (4), 3724–3733.
Li, M., Wilkins, M.R., 2020. Recent advances in polyhydroxyalkanoate production:
feedstocks, strains and process developments. International journal of biological
macromolecules 156, 691–703.
Liu, L.-Y., Xie, G.-J., Xing, D.-F., Liu, B.-F., Ding, J., Ren, N.-Q., 2020. Biological
conversion of methane to polyhydroxyalkanoates: current advances, challenges, and
perspectives. Environmental Science and Ecotechnology 2, 100029.
López, J.C., Arnáiz, E., Merchán, L., Lebrero, R., Muñoz, R., 2018. Biogas-based
polyhydroxyalkanoates production by Methylocystis hirsuta: a step further in
anaerobic digestion biorefineries. Chemical Engineering Journal 333, 529–536.
Lovley, D.R., 2011. Powering microbes with electricity: direct electron transfer from
electrodes to microbes. Environmental microbiology reports 3 (1), 27–35.
Löwe, H., Hobmeier, K., Moos, M., Kremling, A., Pflüger-Grau, K., 2017.
Photoautotrophic production of polyhydroxyalkanoates in a synthetic mixed culture
of Synechococcus elongatus cscB and Pseudomonas putida cscAB. Biotechnology for
biofuels 10 (1), 1–11.
Luangthongkam, P., Laycock, B., Evans, P., Lant, P., Pratt, S., 2019a. Thermophilic
production of poly (3-hydroxybutyrate-co-3-hydrovalerate) by a mixed methane-
utilizing culture. New biotechnology 53, 49–56.
Luangthongkam, P., Strong, P.J., Mahamud, S.N.S., Evans, P., Jensen, P., Tyson, G.,
Laycock, B., Lant, P.A., Pratt, S., 2019b. The effect of methane and odd-chain fatty
acids on 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) synthesis by a
Methylosinus-dominated mixed culture. Bioresources and Bioprocessing 6 (1), 1–10.
Market, M., 2019. Polyhydroxyalkanoate (PHA) Market by Type (Short Chain Length,
Medium Chain Length), Production Method (Sugar Fermentation, Vegetable Oil
Fermentation, Methane Fermentation). Application, and Region-Global Forecast to
2024. Markets and Markets Research Private Ltd.
Marlina, Dian, Sato, Harumi, Hoshina, Hiromichi, Ozaki, Yukihiro, 2018. Intermolecular
interactions of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (P (HB-co-HV)) with
PHB-type crystal structure and PHV-type crystal structure studied by low-frequency
Raman and terahertz spectroscopy. Polymer 135, 331–337.
Matsumoto, K.i., Kitagawa, K., Jo, S.-J., Song, Y., Taguchi, S. 2011. Production of poly
(3-hydroxybutyrate-co-3-hydroxyvalerate) in recombinant Corynebacterium
glutamicum using propionate as a precursor. Journal of biotechnology, 152(4), 144-
146.
Meereboer, K.W., Misra, M., Mohanty, A.K., 2020. Review of recent advances in the
biodegradability of polyhydroxyalkanoate (PHA) bioplastics and their composites.
Green Chemistry 22 (17), 5519–5558.
Mezzolla, V., D’Urso, O.F., Poltronieri, P., 2018. Role of PhaC type I and type II enzymes
during PHA biosynthesis. Polymers 10 (8), 910.
Mittermair, S., Richter, J., Doppler, P., Trenzinger, K., Nicoletti, C., Forsich, C.,
Spadiut, O., Herwig, C., Lackner, M., 2021. Impact of exoD gene knockout on the
polyhydroxybutyrate overaccumulating mutant Mt_a24. International Journal of
Biobased Plastics 3 (1), 1–18.
Miyahara, Y., Yamamoto, M., Thorbecke, R., Mizuno, S., Tsuge, T., 2020. Autotrophic
biosynthesis of polyhydroxyalkanoate by Ralstonia eutropha from non-combustible
gas mixture with low hydrogen content. Biotechnology letters 42 (9), 1655–1662.
Mongili, B., Fino, D., 2021. Carbon monoxide fermentation to bioplastic: the effect of
substrate adaptation on Rhodospirillum rubrum. Biomass Conversion and Biorefinery
11 (2), 705–714.
Monshupanee, T., Incharoensakdi, A., 2014. Enhanced accumulation of glycogen, lipids
and polyhydroxybutyrate under optimal nutrients and light intensities in the
cyanobacterium Synechocystis sp. PCC 6803. Journal of applied microbiology 116
(4), 830–838.
Moreira, J.P., Diender, M., Arantes, A.L., Boeren, S., Stams, A.J., Alves, M.M., Alves, J.I.,
Sousa, D.Z., 2021. Propionate production from carbon monoxide by synthetic co-
cultures of Acetobacterium wieringae spp. and propionigenic bacteria. Applied and
Environmental Microbiology.
Morgan-Sagastume, F., Valentino, F., Hjort, M., Cirne, D., Karabegovic, L., Gerardin, F.,
Johansson, P., Karlsson, A., Magnusson, P., Alexandersson, T., 2014.
Polyhydroxyalkanoate (PHA) production from sludge and municipal wastewater
treatment. Water Science and Technology 69 (1), 177–184.
14
Bioresource Technology 344 (2022) 126307
Mozumder, M.S.I., Garcia-Gonzalez, L., De Wever, H., Volcke, E.I., 2015. Poly (3-
hydroxybutyrate)(PHB) production from CO2: model development and process
optimization. Biochemical Engineering Journal 98, 107–116.
Myung, J., Flanagan, J.C., Waymouth, R.M., Criddle, C.S., 2017. Expanding the range of
polyhydroxyalkanoates synthesized by methanotrophic bacteria through the
utilization of omega-hydroxyalkanoate co-substrates. AMB Express 7 (1), 1–10.
Myung, J., Flanagan, J.C., Waymouth, R.M., Criddle, C.S., 2016a. Methane or methanol-
oxidation dependent synthesis of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) by
obligate type II methanotrophs. Process Biochemistry 51 (5), 561–567.
Myung, J., Kim, M., Pan, M., Criddle, C.S., Tang, S.K., 2016b. Low energy emulsion-
based fermentation enabling accelerated methane mass transfer and growth of poly
(3-hydroxybutyrate)-accumulating methanotrophs. Bioresource technology 207,
302–307.
Nangle, S.N., Ziesack, M., Buckley, S., Trivedi, D., Loh, D.M., Nocera, D.G., Silver, P.A.,
2020. Valorization of CO2 through lithoautotrophic production of sustainable
chemicals in Cupriavidus necator. Metabolic Engineering 62, 207–220.
Nguyen, T.T., Lee, E.Y., 2021. Methane-based biosynthesis of 4-hydroxybutyrate and P
(3-hydroxybutyrate-co-4-hydroxybutyrate) using engineered Methylosinus
trichosporium OB3b. Bioresource Technology 335, 125263.
Orita, I., Nishikawa, K., Nakamura, S., Fukui, T., 2014. Biosynthesis of
polyhydroxyalkanoate copolymers from methanol by Methylobacterium extorquens
AM1 and the engineered strains under cobalt-deficient conditions. Applied
microbiology and biotechnology 98 (8), 3715–3725.
Panda, B., Mallick, N., 2007. Enhanced poly-β-hydroxybutyrate accumulation in a
unicellular cyanobacterium, Synechocystis sp. PCC 6803. Letters in applied
microbiology 44 (2), 194–198.
Panich, J., Fong, B., Singer, S.W., 2021. Metabolic Engineering of Cupriavidus necator
H16 for Sustainable Biofuels from CO2. Trends in biotechnology 39 (4), 412–424.
Park, I., Jho, E.H., Nam, K., 2014. Optimization of carbon dioxide and valeric acid
utilization for polyhydroxyalkanoates synthesis by Cupriavidus necator. Journal of
Polymers and the Environment 22 (2), 244–251.
Pérez, R., Cantera, S., Bordel, S., García-Encina, P.A., Muñoz, R., 2019. The effect of
temperature during culture enrichment on methanotrophic polyhydroxyalkanoate
production. International Biodeterioration & Biodegradation 140, 144–151.
Pradani, L., Rohman, M.S., Margino, S., 2020. The structural insight of class III of
polyhydroxyalkanoate synthase from Bacillus sp. PSA10 as revealed by in silico
analysis. Indonesian. Journal of Biotechnology 25 (1), 33–42.
Ray, S., Kalia, V.C., 2017. Microbial cometabolism and polyhydroxyalkanoate co-
polymers. Indian journal of microbiology 57 (1), 39–47.
Reichardt, R., Rieger, B., 2011. Poly (3-hydroxybutyrate) from carbon monoxide.
Synthetic Biodegradable Polymers 49–90.
Revelles, O., Beneroso, D., Menendez, J.A., Arenillas, A., García, J.L., Prieto, M.A., 2017.
Syngas obtained by microwave pyrolysis of household wastes as feedstock for
polyhydroxyalkanoate production in Rhodospirillum rubrum. Microbial biotechnology
10 (6), 1412–1417.
Revelles, O., Tarazona, N., García, J.L., Prieto, M.A., 2016. Carbon roadmap from syngas
to polyhydroxyalkanoates in Rhodospirillum rubrum. Environmental microbiology 18
(2), 708–720.
Rodríguez, Y., Firmino, P.I.M., Arnáiz, E., Lebrero, R., Muñoz, R., 2020a. Elucidating the
influence of environmental factors on biogas-based polyhydroxybutyrate production
by Methylocystis hirsuta CSC1. Science of The Total Environment 706, 135136.
Rodríguez, Y., Firmino, P.I.M., Pérez, V., Lebrero, R., Muñoz, R., 2020b. Biogas
valorization via continuous polyhydroxybutyrate production by Methylocystis hirsuta
in a bubble column bioreactor. Waste Management 113, 395–403.
Roh, H., Lee, J.S., Choi, H.I., Sung, Y.J., Choi, S.Y., Woo, H.M., Sim, S.J., 2021. Improved
CO2-derived polyhydroxybutyrate (PHB) production by engineering fast-growing
cyanobacterium Synechococcus elongatus UTEX 2973 for potential utilization of flue
gas. Bioresource technology 327, 124789.
Sagong, H.-Y., Son, H.F., Choi, S.Y., Lee, S.Y., Kim, K.-J., 2018. Structural insights into
polyhydroxyalkanoates biosynthesis. Trends in biochemical sciences 43 (10),
790–805.
Salem, R., Soliman, M., Fergala, A., Audette, G.F., ElDyasti, A., 2021. Screening for
methane utilizing mixed communities with high polyhydroxybutyrate (Phb)
production capacity using different design approaches. Polymers 13 (10), 1579.
Sarma, M.K., Kaushik, S., Goswami, P., 2016. Cyanobacteria: A metabolic power house
for harvesting solar energy to produce bio-electricity and biofuels. Biomass and
Bioenergy 90, 187–201.
Schäfer, C., Friedrich, B., Lenz, O., 2013. Novel, oxygen-insensitive group 5 [NiFe]-
hydrogenase in Ralstonia eutropha. Applied and environmental microbiology 79 (17),
5137–5145.
Schoelmerich, M.C., Müller, V., 2019. Energy conservation by a hydrogenase-dependent
chemiosmotic mechanism in an ancient metabolic pathway. Proceedings of the
National Academy of Sciences 116 (13), 6329–6334.
Schubert, P., Steinbüchel, A., Schlegel, H.G., 1988. Cloning of the Alcaligenes eutrophus
genes for synthesis of poly-beta-hydroxybutyric acid (PHB) and synthesis of PHB in
Escherichia coli. Journal of bacteriology 170 (12), 5837–5847.
Schuchmann, K., Müller, V., 2014. Autotrophy at the thermodynamic limit of life: a
model for energy conservation in acetogenic bacteria. Nature Reviews Microbiology
12 (12), 809–821.
Shah, N.N., Hanna, M.L., Taylor, R.T., 1996. Batch cultivation of Methylosinus
trichosporium OB3b: V. Characterization of poly-β-hydroxybutyrate production under
methane-dependent growth conditions. Biotechnology and bioengineering 49 (2),
161–171.
Shi, Q., Tang, J., Liu, R., Wang, L., 2021. Toxicity in vitro reveals potential impacts of
microplastics and nanoplastics on human health: A review. Critical Reviews in
Environmental Science and Technology 1–33.
J. Yoon and M.-K. Oh
Shin, S.K., Ko, Y.J., Lee, M.-E., You, S.K., Hyeon, J.E., Han, S.O., 2021. Enhanced
production of polyhydroxybutyrate from syngas by using nanoscaled cellulose
particles with a syngas-converting enzyme complex immobilized on Ralstonia
eutropha. Journal of Cleaner Production 285, 124903.
Singh, M.K., Rai, P.K., Rai, A., Singh, S., Singh, J.S., 2019. Poly-β-hydroxybutyrate
production by the cyanobacterium Scytonema geitleri Bharadwaja under varying
environmental conditions. Biomolecules 9 (5), 198.
Taepucharoen, K., Tarawat, S., Puangcharoen, M., Incharoensakdi, A., Monshupanee, T.,
2017. Production of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) under
photoautotrophy and heterotrophy by non-heterocystous N2-fixing cyanobacterium.
Bioresource technology 239, 523–527.
Taga, Naohiko, Tanaka, Kenji, Ishizaki, Ayaaki, 1997. Effects of rheological change by
addition of carboxymethylcellulose in culture media of an air-lift fermentor on poly-
D-3-hydroxybutyric acid productivity in autotrophic culture of hydrogen-oxidizing
bacterium. Alcaligenes eutrophus. Biotechnology and bioengineering 53 (5), 529–533.
Tamis, J., Lužkov, K., Jiang, Y., van Loosdrecht, M.C., Kleerebezem, R., 2014.
Enrichment of Plasticicumulans acidivorans at pilot-scale for PHA production on
industrial wastewater. Journal of Biotechnology 192, 161–169.
Tan, G.-Y.A., Chen, C.-L., Li, L., Ge, L., Wang, L., Razaad, I.M.N., Li, Y., Zhao, L., Mo, Y.,
Wang, J.-Y., 2014. Start a research on biopolymer polyhydroxyalkanoate (PHA): a
review. Polymers 6 (3), 706–754.
Tanaka, K., Ishizaki, A., Kanamaru, T., Kawano, T., 1995. Production of poly (D-3-
hydroxybutyrate) from CO2, H2, and O2 by high cell density autotrophic cultivation
of Alcaligenes eutrophus. Biotechnology and bioengineering 45 (3), 268–275.
Tanaka, K., Miyawaki, K., Yamaguchi, A., Khosravi-Darani, K., Matsusaki, H., 2011. Cell
growth and P (3HB) accumulation from CO2 of a carbon monoxide-tolerant
hydrogen-oxidizing bacterium, Ideonella sp. O-1. Applied microbiology and
biotechnology 92 (6), 1161–1169.
Tanaka, K., Mori, S., Hirata, M., Matsusaki, H., 2016. Autotrophic growth of Paracoccus
denitrificans in aerobic condition and the accumulation of biodegradable plastics
from CO2. Environ Ecol Res 4, 231–236.
Tarawat, Somchai, Incharoensakdi, Aran, Monshupanee, Tanakarn, 2020.
Cyanobacterial production of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) from
carbon dioxide or a single organic substrate: improved polymer elongation with an
extremely high 3-hydroxyvalerate mole proportion. Journal of Applied Phycology 32
(2), 1095–1102.
Verma, R., Vinoda, K., Papireddy, M., Gowda, A., 2016. Toxic pollutants from plastic
waste-a review. Procedia Environmental Sciences 35, 701–708.
Vermaas, W.F. 2001. Photosynthesis and respiration in cyanobacteria. e LS, https://doi.
org/10.1038/npg.els.0001670.
15
Bioresource Technology 344 (2022) 126307
Vidra, A., Németh, Á., 2018. Bio-produced acetic acid: a review. Periodica Polytechnica
Chemical Engineering 62 (3), 245–256.
Vigneswari, S., Noor, M.S.M., Amelia, T.S.M., Balakrishnan, K., Adnan, A., Bhubalan, K.,
Amirul, A.-A.A., Ramakrishna, S., 2021. Recent Advances in the Biosynthesis of
Polyhydroxyalkanoates from Lignocellulosic Feedstocks. Life 11 (8), 807.
Volova, T., Zhila, N., Shishatskaya, E., 2015. Synthesis of poly (3-hydroxybutyrate) by
the autotrophic CO-oxidizing bacterium Seliberia carboxydohydrogena Z-1062.
Journal of Industrial Microbiology and Biotechnology 42 (10), 1377–1387.
Volova, T.G., Kiselev, E.G., Shishatskaya, E.I., Zhila, N.O., Boyandin, A.N.,
Syrvacheva, D.A., Vinogradova, O.N., Kalacheva, G.S., Vasiliev, A.D., Peterson, I.V.,
2013. Cell growth and accumulation of polyhydroxyalkanoates from CO2 and H2 of a
hydrogen-oxidizing bacterium, Cupriavidus eutrophus B-10646. Bioresource
technology 146, 215–222.
Weiss, T.L., Young, E.J., Ducat, D.C., 2017. A synthetic, light-driven consortium of
cyanobacteria and heterotrophic bacteria enables stable polyhydroxybutyrate
production. Metabolic engineering 44, 236–245.
Wendlandt, K.-D., Jechorek, M., Helm, J., Stottmeister, U., 2001. Producing poly-3-
hydroxybutyrate with a high molecular mass from methane. Journal of
biotechnology 86 (2), 127–133.
Wilkins, M.R., Atiyeh, H.K., 2011. Microbial production of ethanol from carbon
monoxide. Current Opinion in Biotechnology 22 (3), 326–330.
Yoon, J., Chang, W., Oh, S.-H., Choi, S.-H., Yang, Y.-H., Oh, M.-K., 2021. Metabolic
engineering of Methylorubrum extorquens AM1 for poly (3-hydroxybutyrate-co-3-
hydroxyvalerate) production using formate. International Journal of Biological
Macromolecules 177, 284–293.
Zaldívar Carrillo, J.A., Stein, L.Y., Sauvageau, D., 2018. Defining nutrient combinations
for optimal growth and polyhydroxybutyrate production by Methylosinus
trichosporium OB3b using response surface methodology. Frontiers in microbiology
9, 1513.
Zhang, T., Wang, X., Zhou, J., Zhang, Y., 2018. Enrichments of
methanotrophic–heterotrophic cultures with high poly-β-hydroxybutyrate (PHB)
accumulation capacities. Journal of Environmental Sciences 65, 133–143.
Zhu, B., van Dijk, G., Fritz, C., Smolders, A.J., Pol, A., Jetten, M.S., Ettwig, K.F., 2012.
Anaerobic oxidization of methane in a minerotrophic peatland: enrichment of
nitrite-dependent methane-oxidizing bacteria. Applied and environmental
microbiology 78 (24), 8657–8665.
Zuñiga, C., Morales, M., Revah, S., 2013. Polyhydroxyalkanoates accumulation by
Methylobacterium organophilum CZ-2 during methane degradation using citrate or
propionate as cosubstrates. Bioresource technology 129, 686–689.