International Journal of Biological Macromolecules 89 (2016) 161–174

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International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

Review

Microbial production of polyhydroxyalkanoates (PHAs) and its

copolymers: A review of recent advancements
Anbreen Anjum, Mohammad Zuber ∗ , Khalid Mahmood Zia ∗ , Aqdas Noreen,
Muhammad Naveed Anjum, Shazia Tabasum
Institute of Chemistry, Government College University, Faisalabad 38030, Pakistan

a r t i c l e i n f o a b s t r a c t

Article history: Traditional mineral oil based plastics are important commodity to enhance the comfort and quality of
Received 22 March 2016 life but the accumulation of these plastics in the environment has become a major universal problem due
Received in revised form 15 April 2016 to their low biodegradation. Solution to the plastic waste management includes incineration, recycling
Accepted 22 April 2016
and landfill disposal methods. These processes are very time consuming and expensive. Biopolymers
Available online 25 April 2016
are important alternatives to the petroleum-based plastics due to environment friendly manufacturing
processes, biodegradability and biocompatibility. Therefore use of novel biopolymers, such as polylactide,
Keywords:
polysaccharides, aliphatic polyesters and polyhydroxyalkanoates is of interest. PHAs are biodegradable
Biodegradable polymers
Polyhydoxyalkanoates
polyesters of hydroxyalkanoates (HA) produced from renewable resources by using microorganisms
Economical production as intracellular carbon and energy storage compounds. Even though PHAs are promising candidate for
Industrial applications biodegradable polymers, however, the production cost limit their application on an industrial scale. This
Recent advances article provides an overview of various substrates, microorganisms for the economical production of
PHAs and its copolymers. Recent advances in PHAs to reduce the cost and to improve the performance
of PHAs have also been discussed.
© 2016 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
2. Structures and classification of PHA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .162
3. Physical properties of PHA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
3.1. Properties of short-chain-length and medium-chain-length PHA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
3.2. Properties of P (3HB) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
3.3. Properties of P (4HB) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
3.4. Properties of P (3HB-co-3HV) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
3.5. Properties of PHA terpolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4. Biosynthesis of PHAs and its copolymer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4.1. Sources of PHAs and its copolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4.2. Microorganisms used for economical production of PHAs and its copolymer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4.3. Substrates used for economical production of PHAs and its copolymer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
4.4. Biosynthetic pathways . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
5. Biodegradability and biocompatibility of PHAs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
6. Industrial PHAs and their applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
6.1. Biopol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
6.2. Nodax . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
6.3. Degra pol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
6.4. Biogreen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169

∗ Corresponding authors.
E-mail address: mohammadzuber@gmail.com (M. Zuber).

http://dx.doi.org/10.1016/j.ijbiomac.2016.04.069
0141-8130/© 2016 Elsevier B.V. All rights reserved.
162 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174

7. Recent advances in PHAs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169

7.1. Blends of PHAs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
7.2. Nanocomposites of polyhydroxyalkanoates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
8. Concluding remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171

1. Introduction a minor constituent were explained [16]. Today, approximately

150 different monomer constituents of polyhydroxyalkanoates,
Petroleum-derived plastics (Traditional mineral oil based plas- which posses straight, branched, saturated, unsaturated and aro-
tics) have highly versatile qualities of lightness, robustness, matic structures and over 90 genera of microbial species have been
durability and resistance to degradation. They have become valu- reported to accumulate these polyesters [17,18].
able materials for modern life successfully replacing many other
substances over the years with a wide range of applications in 2. Structures and classification of PHA
domestic, medical and industrial fields in the form of disposable
gears, packaging, furniture, machinery frames and accessories to PHAs have general formula [19] shown in Fig. 1. Struc-
enhance the quality and comfort in life [1]. Yearly about 150 mil- turally PHAs family can be classified into two groups; short
lion tonnes, plastic materials have been consumed worldwide [2]. chain length or medium chain length polyhydroxyalkanoates.
In last few decades, progression in plastic consumption is expected This classification is based on the number of carbon atoms in
to continue until 2020 [3]. the chain of the branching polymers that range from 3 to 14
Large-scale dependence on petroleum-derived plastics has carbon atoms and type homopolymers or heteropolymers pro-
resulted in rapid reduction of crude oil and serious pollution ducing monomeric units [20]. Short-chain lengths PHAs (PHASCL)
problems. Methods used for the disposal of plastic materials are consist of 3–5 carbon atoms. Examples of this class are poly
challenging. In landfills, the degradation rates are tremendously (3-hydroxybutyrate) P(3HB), poly(4-hydroxybutyrate) P(4HB) and
slow. Incineration generates toxic by-products and expensive. poly(3-hydroxyvalerate) P(3HV) or the copolymer P(3HB-co-3HV).
Recycling can be done but is very time-consuming process and also Medium-chain lengths PHAs (PHAMCL) consist of 6–14 or more
cause alteration in the properties of plastic materials. The sorting than 14 carbon atoms [21]. Examples include homopolymers
of a wide range of discarded plastic material is very difficult. Fur- poly (3-hydroxyhexanoate) P(3HHx), poly(3-hydroxyoctanoate)
thermore, the presence of additives such as coatings, fillers and P(3HO) and copolymers such as P(3HHx-co-3HO) [22].
pigments limit the use of the recycled material. Thus, to decrease Difference between two classes is mainly due to the substrate
the environmental impacts of plastics is to replace conventional specificity of the PHA synthases that can accept 3HAs of a cer-
petroleum-derived polymers with biopolymers such as polylactide, tain range of carbon length. For example, PHA synthase of A.
polysaccharides, aliphatic polyesters and polyhydroxyalkanoates eutrophus can polymerise 3HAs consisting of 3–5 carbon atoms
that possess similar physicochemical properties as conventional whereas PHA synthase of Pseudomonas oleovorans can only accept
plastics [4]. 3HAs of 6–14 carbon atoms [23]. Hybrid polymers comprising both
Biopolymers are important alternatives to petroleum-derived short-chain and medium-chain monomeric units also exist, such
polymers due to their biodegradability, eco-friendly manufacturing as poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [4]. Because
processes and enormous ranges of applications in various sectors of the stereo specificity of the biosynthetic enzymes, all monomers
from consumable goods to medical field. They can be produced are in R-configuration. Monomers with various functional groups,
through bio-refineries as part of integrated bioprocesses. Natu- such as halogen, hydroxy, epoxy, cyano, carboxyl and esterified car-
rally biopolymers are produced by variety of microorganisms and boxyl groups on the chain have also been discovered in mcl-PHAs
plants. Most of the biopolymers are biocompatible; they have no [24].
adverse effects on biological systems. It is believed that biopoly- Production of PHB homopolymer has been resulted by
mers of bacterial origin are produced either as a result of their the utilization of even numbered n-alkanoates whereas odd
defence mechanism or as storage material. In both cases, bacteria numbered n-alkanoates produce copolymers of 3HB and 3-
decompose biopolymers [5,6]. hydroxyvalerate (3HV). Terpolymers of poly(3-hydroxybutyrate-
Biodegradable plastics such as polyhydroxyalkanoates (PHAs) co-3-hydroxyvalerate-co-3-hydroxyheptanoate) have been pro-
are more promising due to their inherent biodegradability, sus- duced from alkanoic acids of odd carbon numbers by recombinant
tainability and environment-friendly properties [7]. PHAs are 100% strain of A. eutrophus harboring the PHA synthase gene of
biodegradable polymers. They are thermoplastic polyesters of sev- Aeromonas caviae [25,26].
eral R-hydroxyalkanoic acids, which are produced as carbon and PHAs are accumulated as granular inclusions in cell cytoplasm
energy reserves or reducing power storage materials by numer- that are typically 0.2 ± 0.5 mm in diameter and can be visualized
ous microorganisms (Gram-negative and Gram-positive bacteria) with a phase contrast light microscope due to their high refractiv-
in the presence of excess carbon, especially when another essen- ity or using staining dyes such as Sudan Black B, the oxazine dye
tial nutrient such as oxygen, nitrogen or phosphorus is limiting or Nile Blue A or Nile red. The molecular weight of these compounds
after pH shifts [8–11]. When a limiting nutrient is provided to the ranges from 2 × 105 to 3 × 105 Da, depending on the type of micro-
cell, these energy storage compounds are degraded and are used bial species and the growth conditions such as pH, fermentation
for bacterial growth as carbon source.
Polyhydroxybutyrate (PHB) was the first PHA to be identified
in 1926 by Maurice Lemoige in the bacterium Bacillus megaterium, R O
which showed intra-cellular granules [12]. PHB is the most widely
studied and best-characterized member of PHAs that accumulated O CH (CH2)n C
upto 80% of the cell dry weight [13,14]. The presence of 3HB as a
100-30,000
PHA monomer was reported in activated sludge [15]. The presence
of 3HV as a major constituent and 3-hydroxyhexanoate (3HHx) as Fig. 1. General Formula of PHA [19].
 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174
conditions, modes of fermentation (batch, fed-batch, continuous)
and type and concentration of the carbon source [23].
3. Physical properties of PHA
PHAs vary considerably in their physical and chemical charac-
teristics. Polymer hydrophobicity, melting point, glass transition
temperature and degree of crystallinity utterly depend on the com-
position of the monomer [3]. The family of PHAs exhibits a wide
variety of mechanical properties from hard crystalline to elastic.
3.1. Properties of short-chain-length and medium-chain-length
PHA
Short-chain-length biopolymers are stiff, brittle and possess
a high degree of crystallinity in the range of 60–80% whereas
medium-chain-length PHA is flexible and elastic materials with low
crystallinity (25%), low tensile strength, high elongation to break,
low melting temperatures and Tg below room temperature [27,28].
3.2. Properties of P (3HB)
Since PHB is the most widely studied polymer so the proper-
ties of PHAs have been explained by taking it into consideration.
P (3HB) is highly crystalline due to its stereo regularity. P (3HB)
is water insoluble and relatively resistant to hydrolytic degrada-
tion. It has low O2 permeability and good thermoplastic properties
with poor mechanical properties like Young’s modulus and ten-
sile strength than those of petroleum-based polymers for example
polypropylene [28]. The densities of crystalline and amorphous
PHB are 1.26 and 1.18 g/cm3 , respectively. The molecular weight
of P (3HB) produced from wild-type bacteria is usually in the
range of 10,000–3,000,000 Da with a poly dispersity of around 2.
P (3HB) is optically pure and possesses piezo electricity; this helps
in the process of inducing osteogenesis [9]. P (3HB) is highly brit-
tle and stiff material. It was found that the physical properties of P
(3HB) homopolymer could be improved by producing high molec-
ular weight polymers. This polymer was produced by heterologous
expression of cloned PHA biosynthesis gene from C. necator in
recombinant Escherichia coli.
3.3. Properties of P (4HB)
Poly (4-hydroxybutyrate) P(4HB), on the other hand, is a strong
and malleable thermoplastic material with a tensile strength
closely comparable to that of polyethylene. It has an elongation at
break of 100% resulting in extremely elastic properties. When com-
bined with other hydroxyacids, the material properties of P (4HB)
can be varied [29].
3.4. Properties of P (3HB-co-3HV)
The incorporation of secondary different HA monomers other
than 3HB such as 3-hydroxyvalerate, 3-hydroxyhexanoate, 3-
hydroxypropionate and 4-hydroxybutyrate into the polymer chain
to form copolymers is a common strategy. It greatly improves
the material properties of P (3HB) such as crystallinity, melting
point, stiffness and toughness. The most well-known copolymer
P (3HB-co-3HV) has lower crystallinity, lower melting temper-
ature, decreased stiffness, increased elongation to break, being
more flexible (decrease in Young’s Modulus), and tougher (increase
in impact strength) than P(3HB). The mole percentage of 3HV is
important in determining the properties of the copolymer. Thermo
mechanical properties can be widely varied because the com-
position of P (3HB-co-3HV) can range from 0 to 30 mol% 3HV
[24]. For example P (3HB-co-3HV) containing more than 20 mol%
163
of 3HV units can be used to make films and fibers with dif-
ferent elasticities by controlling the processing conditions [22].
Poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) combines the
thermo-mechanical properties of PE (strength, flexibility, ductil-
ity, toughness, elasticity) with the physical-chemical properties of
polyesters (printability, dyeability). It can form blends with PLA and
thermoplastic starch.
3.5. Properties of PHA terpolymers
PHA terpolymers are known to be better materials compared
to copolymers. Introduction of more than one secondary monomer
is assumed to result in improved properties. It has been demon-
strated that P (3HB-co-3HV-co-3HHx) terpolymer with higher
amorphousness could be produced. No melting peak was detected
in terpolymer with 39 mol% 3HV and 3 mol% 3HHx, while it shows
tensile strength of 12 MPa and elongation at break 408%. P (3HB-
co-3HV-co-4HB) with 93 mol% 4HB and 3 mol% 3HV exhibited
elongation of 430%, toughness of 33 MPa and Young’s Modulus of
127 MPa, which is similar to LDPE, while terpolymer with 55 mol%
4HB and 34 mol% 3HV showed a Young’s Modulus of 618 MPa, sim-
ilar to that of polypropylene [30,31].
Table 1 gives a comparison of the properties of different PHAs
with conventional petroleum-based polymers. Table 2 gives prop-
erties of commercial PHAs.
4. Biosynthesis of PHAs and its copolymer
4.1. Sources of PHAs and its copolymers
PHAs are a family of biodegradable polymers produced by
a variety of Gram-positive and Gram-negative bacteria. Over
300 species, mainly of bacteria are reported to produce these
polymers [39,40]. These bacteria can accumulate biopolymers in
their cytoplasm as an intracellular carbon and energy reserve
[41–44]. Bacteria that are used for the production of PHAs can be
divided into two groups based on the stress conditions required for
PHA synthesis. The first group of bacteria requires the limitation of
an essential nutrient such as nitrogen, phosphorous, magnesium or
sulphur with an excess of carbon source. The bacteria included in
this group are A. eutrophus, Protomonas extorquens and Protomonas
oleovorans. The second group of bacteria, which include Alcaligenes
latus, a mutant strain of Azotobacter vinelandii and recombinant
E. coli do not require nutrient limitation for PHA synthesis [45,46].
4.2. Microorganisms used for economical production of PHAs and
its copolymer
Among the bacterial species that are used to accumu-
late large amounts of PHAs and its copolymers are wild and
recombinant strains of Ralstonia eutropha formerly called A.
eutrophus, Wautersia eutropha or Cupriavidus necator [47–79],
Azotobacter sp. [80–99], Bacillus sp. [100–114], Pseudomonas sp.
[115–145], Burkholderia sp. [146–150], Halomonas sp. [151–154],
Haloferax sp. [155,156], Aeromonas sp. [157–161], Methylobac-
terium sp. [162–167], Thermus thermophilus [168], Hydrogenophaga
pseudoflava [169,170], Saccharophagus degradans [171], Coma-
monas sp. [172–178], A. latus also known as Azohydromonas lata
[179–184], Rhodobacter sphaeroides [185–188], Zobellella denitri-
ficans [189], Activated sludge [190–203], Cyanobacteria [204],
Chromobacterium sp. [205–207], Erwinia sp. [208], recombinant E.
coli. [209–229].
164 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174

Table 1
Comparison of the Physical Properties of different PHAs with conventional petroleum-based polymers [28,32–36].

Sr # Polymer Copolymer Young’s Modulus Tensile strength Elongation at Melting Glass Transition
content GPa MPa Break% Temperature ◦ C Temperature ◦ C

1 P(3HB) 3.5–4 40 3–8 173–180 5–9

2 P(4HB) 149 104 1000 53 −50
3 P(3HB-co-3HV) (3 mol% 3HV) 2.9 38 n.a 170 n.a
(9 mol% 3HV) 1.9 37 n.a 162 n.a
(14 mol% 3HV) 1.5 35 n.a 150 n.a
(20 mol% HV) 1.2 32 n.a 145 −1
(25 mol% 3HV) 0.7 30 n.a 137 n.a
4 P(3HB-co-4HB) (3 mol% 4HB) n.a 28 45 166 n.a
(10 mol% 4HB) n.a 24 242 159 n.a
(16 mol% 4HB) n.a 26 444 130 −7
(64 mol% 4HB) 30 17 591 50 −35
(90 mol% 4HB) 100 65 1080 50 −42
5 P(3HHx-co-3HO) n.a 10 300 61 n.a
6 P(3HB-co-3HA) (6 mol% 3HA) 0.2 17 680 133 −8
7 P(3HB-co-HP) (67 mol% HP) n.a n.a n.a 44 −19
8 P(3HB-co-3HHx) n.a 20 850 52 −4
P(3HO) n.a 6–10 300–450 61 n.a
9 Isotactic polypropylene 1.0–1.7 29.3–38.6 500–900 170–176 −10
10 HDPE 0.4–1.0 17.9–33.1 12–700 112–132 −80
11 LDPE 0.05–0.1 15.2–78.6 150–600 88–130 −36
12 PS 3.0–3.1 50 3–4 80–110 21
13 Nylon-6,6 2.8 83 60 265 50
14 Polyethylene-terethalate 2.2 56 7300 262 3400

Table 2
Properties of Commercial PHAs [37,38].

P(3HB) Biomer P240 P(3HB) Biomer P226 P(3HB-co-3HV) Biopol P(3HB-co-3HHx) Nodax, Kaneka

Physical Properties
MFR (g/10 min) 5–7 9–13 0.1–100
Density (g/cm3 ) 1.17 1.25 1.23–1.26 1.07–1.25
Transparency (%) 0.7 White powder translucent film

Mechanical Properties
Tensile strength at yield (MPa) 18–20 24–27 10–20
Elongation at yield (%) 10–17 6–9 10–25
Flexural modulus (MPa) 1000–1200 1700–2000 40 Several orders of magnitude

Thermal Properties
HDT ◦ C 60–100
Vicat softening point ◦ C 53 96 60–120

4.3. Substrates used for economical production of PHAs and its
copolymer
A wide range of substrates has been used for the produc-
tion of PHAs, its copolymers and terpolymers including industrial
byproducts, fats and oils, lignocellulosic raw materials, agricultural
and household waste materials, glycerol, sugars and wastewater.
Extensive research has been carried out using sugar beet molasses
[80,105,220], sugar cane molasses [101,102,230–232], sugar cane
liquor [233], soy molasses and hydrolysed soy [103,104,128], corn
steep liquor [212], glycerol [55,129,223,234,235], fish peptone
medium [228], casein hydrolysate [55], urea [101], cheese whey
and hydrolysed whey [47,83,61,162,163,216,217,225–227,236],
wheat based bio refinery [48,151], starch [83,106], cellulose
and cellulose hydrolysates [171,237], sucrose [179,238,239],
xylose [116,139,146,147,237], galactose, mannose and rham-
nose [171], arabinose [115], glucose [52,115,138,179,221,240,241]
fructose [178], rice bran and corn starch [156], sugarcane
bagasse hydrolysates [14,147], triacylglycerides (TAG) [158], tal-
low [123,131], lard oil, butter oil and coconut oil [122,130], lactic
acid [49,50,56–58], oleic acid [54], acetic acid [51–56], rapeseed
meal hydrolysates [59], sunflower meal hydrolysates [60], olive
oil [63], corn oil [62,63], linseed oil [242], Brassica carinata oil
[243], palm oil and its products [63,244], coprah oil [245], hazel-
nut oil and sesame oil [246], soybean oil [65], jatropha oil [247],
spent palm oil [248], waste rapeseed oil [71,249], waste vegetable
oil [121], gaseous mixture of H2 , O2 and CO2 [66,67], natural gas
[250], municipal waste waters [68,202,251], starchy wastewater
[69], food scraps [70], waste sesame oil [72], waste frying oil [73],
waste cooking oil [74,118,119], waste potato starch [75], potato
processing waste [252], malt waste [104,107], cottonseed oil [77],
alpechin medium (waste water from olive oil mil) [82,134,253],
swine waste liquor [46], residual oil from rhamnose biotechnology
[64], biodiesel waste water [254], food processing waste efflu-
ents [199,255], brewery waste effluents [256], paper mill waste
water [191], kraft mill waste water [257], palm oil mill efflu-
ents [258], methanol [164], date syrup [110], hydrolysed cassava
starch [113], hydrolysed sago starch [259], crude jerusalem arti-
choke hydrolyzate [260], volatile fatty acids derived from digested
ensiled grass [261]. Agro-industrial wastes, such as sunflower cake,
soy bran, yeast hydrolysate, corn steep liquor and castor oil seeds
containing solid biodiesel residue have also been used for the pro-
duction of PHAs by C. necator [78].
Specialized carbon sources like 4-hydroxybutyric acid, ␥-
butyrolactone and 1,4-butanediol give rise to 4HB monomers along
with 3HB monomer [32]. Fatty acids, such as nonanoic (9:0) and
octanoic (8:0) acids were found to be more toxic to R.eutropha than
volatile fatty acids, such as acetic acid, propionic acid and butyric
acid [53]. A detailed overview of microbial production of PHAs and
its copolymers including specific biopolymers, microbial strains,
substrate and characterization techniques have been presented in
Table 3.
 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174 165

Table 3
An Overview of Microbial Production of PHAs and its Copolymers.

Sr No. Biopolymer Microbial Strain Substrate (Source) Characterization References

Techniques

1 Poly(3-hydroxybutyrate) C. necator DSM 545 and Glucose and Glycerol FTIR, 1 H NMR, DSC [262]
(PHB) Burkholderia sacchari
2 Poly(3-hydroxybutyrate- R. eutropha ATCC 17699 Fructose and NMR [263]
co-4-hydroxybutyrate) ␥-butyrolactone
[P(3HB-co-4HB)]
13
3 Poly(3-hydroxybutyrate- C. necator Spent palm oil C NMR, DSC, FTIR, GC [248]
co-4-hydroxybutyrate),
P(3HB-co-4HB)
4 PHA/PHB SM-P-1S, SM-P-2S, Jatropha biodiesel FTIR, 1 H NMR [264]
SM-P-3S, SM-P-4S and by-product containing
SM-P1M, SM-P-2M, crude glycerol
SM-P-3M, SM-P4M and
SM-P-5M
5 PHA Recombinant E. coli Cheese whey GC [265]
6 P(3HB) homopolymer and R. eutropha H16 and its Soybean oil [266]
P(3HB-co-3HHx) recombinant strain
copolymer
7 Poly(3-hydroxybutyrate), C. necator strain A-04 Refined sugarcane, Brown DSC [267]
PHB, and sugarcane, Rock sugar,
poly(3-hydroxybutyrate- Toddy palm sugar and
co-4-hydroxybutyrate), Coconut palm sugar as
P(3HB-co-4HB) carbon source, Ammonium
sulfate as nitrogen source
8 Polyhydroxyalkanoates W. eutropha Canola oil FTIR, NMR, DSC [268]
(PHAs)
9 PHA Halomonas hydrothermalis Seaweed-derived crude FTIR, 1 H NMR, DSC [269]
levulinic acid (SDCLA),
containing formic acid,
residual sugars and
dissolved minerals
10 Poly-3-hydroxybutyric Pseudomonas Lactose, glucose and DSC [10]
acid (PHB) and hydrogenovora galactose derived from
poly-(3HB-co-3HV) whey permeate
1
11 Poly-[(R)-3- Bacillus firmus NII 0830 Crude glycerol H NMR [270]
hydroxybutyric acid]
(PHB)
12 Poly (3-hydroxybutyrate) Bacillus sp. SV13 Pineapple and sugarcane NMR, DSC [271]
13 Terpolyesters Recombinant Aeromonas Dodecanoic acid and NMR, GPC, DSC, TGA and [272]
P(3HB-co-3HV-co-3HHx) hydrophila 4AK4 harboring propionic acid stress–strain measurement
␤-ketothiolase gene phaA
and NADPH-dependent
acetoacetyl-CoA reductase
gene phaB
14 PHA T. thermophilus HB8 Lactose from whey-based GC and GC/MS, NMR, GPC [168]
media
15 Poly (3-hydroxybutyrate- Comamonas acidovorans Glucose and 1,4-butanediol [178]
co-4-hydroxybutyrate)
[P(3HB-co-4HB)]
copolymers
16 P (3HB-co-3HV) copolymer C. necator H16 Palm kernel oil, crude palm [273]
oil, and palm olein
17 poly(3-hydroxybutyrate- Recombinant C. necator Palm kernel oil DSC [31]
co-3-hydroxyvalerate-co-
3-hydroxyhexanoate)
18 poly(3-hydroxybutyrate) C. necator DSM 545 Soy cake or soy cake XRD, DSC, NMR, FTIR, GPC, [274]
supplemented with 2.5% POM
(m/m) sugarcane molasses.
19 PHA C. necator DSM 545 Waste glycerol SEC [275]
20 PHB W. eutropha Wheat based bio refinery TEM [276]
1
21 Poly(3-hydroxybutyrate- R. eutropha KCTC 2662 Soybean oil and H NMR [42]
co-4-hydroxybutyrate) ␥-butyrolactone
[P(3HB-co-4HB)]
13
22 P(3HP-co-4HB) Recombinant E. coli Glycerol C NMR, DSC, 1 H NMR,GPC [277]
1
23 PHA Massilia albidiflava, M. Glucose or starch H NMR [278]
lutea, M. dura, M. plicata, M.
brevitalea, M. aerilata and
M. aurea
24 [P (3HB-co-3HV)], PHB Bacillus sp. Glucose, glycerol, sod. TEM, GC–MS, 1 H NMR, [279]
acetate XRD, DSC, IR
25 Poly(3-hydroxybutyrate- Cupriavidus sp. USMAA2-4 1,4-butanediol and GPC [280]
co-4-hydroxybutyrate) ␥-butyrolactone
[P(3HB-co-4HB)]
1
26 Poly(3-hydroxybutyrate- Bacillus cereus FA11 Glucose H NMR, FTIR [281]
co-3-hydroxyvalerate)
[poly(3HB-co-3HV)]
166 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174

Table 3 (Continued)

Sr No. Biopolymer Microbial Strain Substrate (Source) Characterization References

Techniques

27 PHA R. eutropha Paddy straw FTIR, Thermogravimetric [282]

MTCC 1472 analysis, DSC
1
28 Poly (3-hydroxybutyrate- Bacillus sp-256, Rhizobium Mahua flower H NMR [283]
co-3-hydroxyvalerate) meliloti, Sphingomonas sp.
[poly(3HB-co-3HV)]
13
29 Poly(␤-hydroxybutyrate- Burkholderia cepacia ATCC Xylose and Levulinic acid C NMR, DSC, 1 H NMR [284]
co-ˇ-hydroxyvalerate) 17759
(P(3HB-co-3HV))
13
30 Poly(3-hydroxybutyrate- W. eutropha harboring the Palm kernel oil, palm olein, C NMR, DSC, 1 H [44]
co-3-hydroxyhexanoate) A. caviae crude palm oil and palm NMR,GPC, TEM
[P(3HB-co-3HHx)] polyhydroxyalkanoate acid oil
(PHA) synthase gene.
31 Poly(3-hydroxyalkanoates) Comamonas testosteroni Castor seed oil, coconut oil, FTIR, 13 C NMR [285]
mustard oil, cotton seed
oil, groundnut oil, olive oil
and sesame oil
13
32 PHB P. fluorescens A2a5 Sugarcane liquor, C NMR [233]
monosodium glutamate
33 PHB B. megaterium Sugarcane molasses and TEM [286]
corn steep liquor
34 PHA Recombinant E. coli Molasses and sucrose DSC, TGA, DTA, GC/MS, TLC [287]
harboring phaC1 of and optical rotation studies
Pseudomonas sp. LDC-5
35 PHB Paracoccus denitrificans Crude glycerol Thermal analysis, GPC [288]
(DSMZ 413) and C. necator
JMP 134
36 Poly (3HB-co-3HV) P. oleovorans (ATCC 29347) Saponified Jatropha curcas (GC/MS), Electrospray [289]
seed oil ionization–time of Flight
mass spectroscopy
(ESI–TOF MS), 1 H, 13 C NMR
and GPC
1
37 P (3HB-co-3HHx) Recombinant C. necator Crude palm kernel oil, H NMR and 13 C NMR, GPC, [290]
strain Re2160/pCB113 jatropha oil, crude palm oil, DSC, TGA
palm olein soybean oil,
corn oil, and coconut oil
38 P (3HB-co-3HA) Recombinant R. eutropha Soybean oil, fructose GPC, DSC [291]
39 PHA P. aeruginosa 42A2 Linseed oil GC/MS, FTIR, 1 H NMR and [242]
13
C NMR, GPC, DSC, TGA,
TEM
40 PHA P. guezennei biovar. tikheau Coprah oil FTIR, 1 H NMR, 13 C NMR, [245]
strain TIK669 (EMBL DSC, GC/MS
AM922198)
41 PHA P. guezennei Glucose, acetate, pyruvate, GC/MS, FTIR, 1 H NMR, 13 C [292]
propionate, valerate, NMR, DSC, GPC-MALLS
hexanoate, heptanoate,
octanoate, decanoate, and
oleic acid
42 mcl-PHAs P. aeruginosa ATCC 27853 B. carinata oil GPC, 1 H NMR, 13 C NMR, ESI [293]
MS, and DSC
43 mcl-PHAs Pseudomonas sp. Gl01 Waste saponified palm oil GC, DSC, GPC, [294]
44 [P (HB-co-HV] B. cepacia ATCC 17759 Spent coffee grounds GC, GPC, DSC, TGA [295]
hydrolyzate (SCGH)
45 Poly (3- hydroxybutyrate) B. sacchari DSM 17165 Wheat straw GC [296]
(P(3HB) lignocellulosic
hydrolysates (WSHs)
1
46 P (3HB-co-4HB) B. sacchari DSM 17165 Wheat straw H NMR, SEC, DSC, TGA [297]
lignocellulosic
hydrolysates (WSHs)
47 mcl PHAs Pseudomonas guezennei Glucose, sodium octanoate, GCMS, NMR, DSC and GPC [298]
and 10-undecenoic acid
48 PHA Pseudomonas putida 27N01 Fluorophenoxyalkanoic GCMS, 1 H NMR, 13 C NMR, [299]
acids DSC, XRD, GPC
1
49 P(3HB-co-3HV) Bacillus circulans MTCC Dextrose H NMR, 13 C NMR, FTIR, [300]
8167 GPC, XRD
50 Poly(3-hydroxybutyrate) Comamonas sp. EB172 Acetic, propionic and TGA, DSC [301]
[P(3HB)] homopolymer butyric acids derived from
and POME
poly(3-hydroxybutyrate-
co-3-hydroxyvalerate)
[P(3HB-co-3HV)]
copolymer
 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174
4.4. Biosynthetic pathways
There are three well-known PHA biosynthetic pathways (Fig. 2).
The monomer composition is related to the carbon source used.
Pathway I is the best known among the PHA biosynthetic pathways
used by C. necator. In this pathway, 3HB monomers are generated
by the condensation of two acetyl-CoA molecules from the tricar-
boxylic acid (TCA) cycle to form acetoacetyl-CoA by the enzyme
␤-ketothiolase. Acetoacetyl CoA reductase acts on acetoacetyl-CoA
to form 3-hydroxybutyryl-CoA. Finally, the PHA synthase enzyme
catalyses the polymerization via esterification of 3 hydroxybutyryl-
CoA into poly(3-hydroxybutyrate) (P(3HB)). Pathways involved
in fatty acid metabolism generate different hydroxyalkanoate
monomers utilized in PHA biosynthesis. Pathway II generates sub-
strates by ␤-oxidation of fatty acids that can be polymerised by the
PHA synthases of Pseudomonads belonging to the ribosomal RNA-
homology group I such as Pseudomonas aeruginosa. In A. caviae, the
␤-oxidation intermediate, trans-2-enoyl-CoA is converted to (R)-
hydroxyacyl-CoA by a (R)-specific enoyl-CoA hydratase. Pathway
III is of significant interest because it helps generate monomers
for PHA synthesis from structurally unrelated and simple, inex-
pensive carbon sources such as glucose, sucrose and fructose. The
(R)-3-hydroxyacyl intermediates from the fatty acid biosynthetic
pathway are converted from their acyl carrier protein (ACP) form to
the CoA form by the enzyme acyl-ACP-CoA transacylase (encoded
by phaG). Fig. 3 shows general scheme for the metabolic pathways
of PHAs synthesis from different carbon sources within a bacte-
rial cell. Fig. 4 represents general PHAs production and extraction
process.
5. Biodegradability and biocompatibility of PHAs
Biodegradability and biocompatibility are important character-
istics of PHAs. An interesting property of polyhydroxyalkanoates
that distinguishes them from traditional petroleum based plastics
is their biodegradability. These biopolyesters are stable in air, inert,
resistant to moisture and are water insoluble [45]. They can be
completely degraded to water and carbon dioxide under aerobic
conditions and to methane and carbon dioxide under anaerobic
conditions by microorganisms in soil, sea, lake water and sewage
[23]. The degrading microorganisms produce extracellular PHA
depolymerases, which convert the polyesters into water-soluble
oligomers and monomers that are used as a carbon source by the
organisms [27]. These extracellular depolymerases are quite well
understood. In addition, most PHA-producing bacteria are able to
degrade the polymer intracellularly. During intracellular degra-
dation, the PHA depolymerase in the cell breaks down P (3HB)
to give 3-hydroxybutyric acid. A dehydrogenase acts on the lat-
ter and oxidises it to acetylacetate and a ␤-ketothiolase acts on
acetylacetate to break it down to acetyl-CoA. The ␤-ketothiolase
enzyme plays an important role in both the biosynthetic and the
biodegradation pathways. Under aerobic conditions, the acetyl-
CoA enters the citric acid cycle and is oxidised to CO2 [303]. Very
little is known about the intracellular depolymerases since they
are always found to be intimately connected to the P(3HB) gran-
ules and the overall process is very complex [304]. They can also be
degraded by thermal means or by enzymatic hydrolysis. In a biolog-
ical system, PHAs can be degraded using microbial depolymerases
as well as by non-enzymatic and enzymatic hydrolysis in animal
tissues. Biodegradation of PHAs is influenced not only by their own
properties, such as stereoregularity, crystallinity i.e. highly crys-
talline materials have lower biodegradability, chemical structure
(especially functional groups and hydrophilicity–hydrophobicity
balance) and accessibility of their surface to PHA-depolymerizing
enzymes, but also depends on a variety of factors including surface
167
area, microbial activity of the disposal environment, pH, temper-
ature, moisture and presence of other nutrient materials [28]. It
has been found that low molecular weights PHAs are more sus-
ceptible to biodegradation. The melting temperature is another
important factor to be considered when studying biodegradation.
As the melting point increases, the biodegradability decreases.
With increasing melting temperature, the enzymatic degradability
decreases [9]. Studies have shown that 85% of PHA was degraded
in seven weeks. PHA has been reported to degrade in aquatic envi-
ronments within 254 days even at temperature not exceeding
60 ◦ C. The degradation of PHA by R. eutropha could occur simul-
taneously with its biosynthesis under nitrogen limitation. This
observation is called “cyclic nature of PHA metabolism”. P (3HB)
has been taken as a prototype in the biodegradation studies of PHAs
(Fig. 5). Microorganisms from the families Pseudonocardiaceae,
Micromonosporaceae, Thermomonosporaceae, Streptosporangiaceae
and Streptomycetaceae predominantly degrade P (3HB) in the envi-
ronment. PHB is compostable over a wide range of temperatures,
even at a maximum of around 60 ◦ C with moisture levels at
55%. P (HB-HV) is biodegraded in microbial active environments
[305–309]. Microorganisms colonize on the surface of the polymer
and secrete enzymes that degrade P (HB-HV) into HB and HV units.
P (3HB-co-3HV) is significantly degraded upon incubation in aero-
bic sewage sludge for 8 weeks. In other studies it was found that P
(3HB-co-3HV) was completely degraded in anaerobic sewage, soil
and sea water within 6, 75 and 350 weeks, respectively [4].
Biocompatibility is characterized by the absence of any toxic
compound generated during polymer degradation, as well as by the
shape and surface porosity of the material. The biocompatibility of
P (3HB) is no surprise, since 3HB is a normal blood constituent and
is found in the cell envelope of eukaryotes [310]. Therefore, PHAs
are potential candidates for use in different medical applications
[18,311].
6. Industrial PHAs and their applications
6.1. Biopol
Biopol was initially manufactured in 1970s by Imperial Chem-
ical Industries ICI, UK. The patents were later sold to Zeneca, then
to Monsanto, and is currently produced by Metabolix (Cambridge,
MA, USA). Biopol is a co-polymer of poly (3-hydroxybutyrate-co-
3-hydroxyvalerate). The copolymer is a thermoplastic and has a
melting point in the range of 140–180 ◦ C. Biopol can be injection
moulded and vacuum formed. It has a range of uses such as, pack-
aging, shampoo bottles, disposable razors, disposable cups, surgical
stitches, surgical pins, disposable knives and forks, woven medical
patches and nappy linings.
6.2. Nodax
Nodax is a recent addition to the PHA copolymer family con-
sisting of 3-hydroxybutyrate and a comparatively small quantity
of medium chain length monomers with side groups greater than
or equal to three carbon units. The mcl units used include 3-
hydroxyhexanoate, 3-hydroxyoctanoate and 3-hydroxydecanoate.
Procter and Gamble have developed this polymer. Nodax is avail-
able as foams, fibers or nonwovens, films and latex among others.
6.3. Degra pol
Degra Pol is a polyester-urethane. It consists of two blocks
of polymers, which impart very different physical and mechan-
ical properties to the final product. It is chemically synthe-
sized from polyhydroxybutyrate-diol (hard segment) and ␣,
168 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174

Fig. 2. PHA Biosynthetic Pathways [9].

 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174 169

␻-dihydroxy-poly (D -caprolactone-block-diethyleneglycol-block-
D -caprolactone) (Soft segment).

6.4. Biogreen

Japan-based Mitsubishi Gas Chemicals developed the produc-

tion of P (3HB) from methanol, and markets it under the trade name
Biogreen.
Industrial PHAs have a wide range of applications. Initially they
were used as packaging materials such as films in bags, cosmetics
containers, shampoo bottles, cups, milk cartons, as latex to cover
cardboards and paper, disposable items, such as razors, utensils,
diapers, feminine hygiene products, pens, combs, bullets, sani-
tary towels, in molding, golf tees, hot-melt and pressure-sensitive
adhesives, as food additives, dairy cream substitutes, non woven
fabrics, binders, thermoformed articles. PHAs can be used to make
foils and diaphragms. Due to its piezoelectric nature, it is possi-
ble to use PHAs for making pressure sensors for keyboards, stretch
and acceleration measuring instruments, material testing, shock
wave sensors, lighters, gas lighters; acoustics: microphone, ultra-
sonic detectors, sound pressure measuring instruments, oscillators,
headphones, loudspeakers, for ultrasonic therapy and atomization
of liquids. The gas barrier property of P(3HB-3HV) is useful for appli-
cations in food packaging and for making plastic beverage bottles.
In addition to potential as a plastic material, PHAs are also useful
as stereo regular compounds which can serve as chiral precur-
sors for the chemical synthesis of optically active compounds. In
medical field PHAs, especially P(3HB), P(3HB-3HV), P(4HB), P(3HO)
and P(3HB-3HHx) are frequently used in tissue engineering. They
are widely used as bone plates, osteosynthetic materials, surgical
sutures, dusting powders, wound dressings, nerve conduits, soft tis-
sue repair, dental and maxillofacial treatment, prodrugs, implants
and tablets. They are useful in the slow release of drugs and hor-
mones. The greatest contribution of PHAs to medicine has been in
the cardiovascular area (pericardial and atrial septal repair patches, Fig. 3. General scheme for the metabolic pathways of PHA synthesis from different
scaffolds for regeneration of arterial tissues, vascular grafts, cardio- carbon sources within a bacterial cell [28].
vascular stents and heart valves).
For agricultural purposes PHAs have been used as mulch films, bonding interactions of poly (3-hydroxybutyrate) (PHB)/chitin
herbicides, insecticides, bacterial inoculants used to enhance nitro- blends were studied [330]. Novel hybrid fibrous membranes
gen fixation in plants. with potential application on the biomedical field were suc-
cessfully developed via electrospinning of blend solutions
7. Recent advances in PHAs of two biocompatible and biodegradable polymers, poly (3-
hydroxybutyrate-co-hydroxyvalerate) (PHBV)/chitosan [331]. Poly
7.1. Blends of PHAs (3-hydroxybutyrate)/cellulose acetate butyrate PHB/CAB blends
were prepared and their miscibilty, crystallization behavior and
Various blends of PHAs have been developed to reduce the phase structure was studied [332]. In another study effects
prices and to improve the performance of PHAs. PHA/PLA blend of hydrogen bond intermolecular interactions on the crys-
is one of the most studied blends as PHB and its copolymer poly tal spherulite of PHB/CAB blends were characterized [333].
(3-hydroxybutyrate-co-hydroxyvalerate) PHBV is very close to PLA Melting, crystallization behavior and phase morphology of
in chemical structure and processing [312–315]. Compatibility of PHB/hydroxyethyl cellulose acetate (HECA) blends was reported
the blends of PHB with poly (methyl methacrylate) (PMMA) and [334]. Phase behavior, crystallization kinetics and tensile prop-
poly (cyclohexyl methacrylate) (PCHMA) were investigated. PHB erties of poly (hydroxybutyrate)/cellulose propionate (PHB/CP)
showed no miscibility with PCHMA. Solubility limit of PHB in blends have been investigated [335]. Environmental degradation
PMMA is 20 wt% [316]. Blends of PHA with rubber [317], poly of Lignin/PHB blends and the role of lignin in either retarding or
(ethylene glycol) (PEG) [318], poly (ethylene oxide) (PEO) [319], accelerating the PHB degradation has been investigated in a study
poly (vinyl alcohol) (PVA) [320], poly (vinyl acetate) (PVAc) [321], [336]. Thermal and mechanical properties of PHB/starch blend films
poly (vinylidene fluoride) [322], poly(2-hydroxyethylmetacrylate) were reported [337].
(PHEMA) [323], poly (vinyl phenol) [324], poly caprolac-
tone [325], poly (butylene succinate) [326], poly (propylene 7.2. Nanocomposites of polyhydroxyalkanoates
carbonate) [327,328] have also been reported. Natural biodegrad-
able polysaccharides, such as cellulose and starch and their PHBV nanocomposites were obtained by the addition of two
derivatives have also been reported to blend with PHA. Morpho- different type of nanoparticles, organically modified platelet
logical and molecular properties of blends of polyhydroxybutyrate- like clay montmorillonite Cloisite 30B (C-30B) and unmod-
hydroxyvalerate (PHB-HV) with maize starch were investigated ified tubular shaped halloysite and studied their effect on
[329]. Crystallization behavior and intermolecular hydrogen the morphology and thermal, mechanical properties of PHBV
170 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174

Fig. 4. General polyhydroxyalkanoates production and extraction process [302].

Fig. 5. Biodegradability process of poly-␤-hydroxybutyrates [204].

nanocomposites. A partially exfoliated structure for PHBV/C-
30B nanocomposites and a good dispersion of HNT in the
PHBV matrix was observed [338]. Mechanical characteristics
of kenaf and lyocell fibre reinforced poly (3-hydroxybutyrate)
composites were compared [339]. Poly(3-hydroxybutyrate-
co-3-hydroxyvalerate)/organo-modified Mg-Al layered double
hydroxide (PHBV/m-LDH) and poly(3-hydroxybutyrate) (PHB)/m-
LDH nanocomposites were synthesized and their structure,
morphology and physical properties were characterized [340].
Mechanical properties, bio-compatibility, and biodegradability
of composites of treated (cross-linked) cellulose acetate (t-CA)
and acrylic acid-grafted poly (hydroxyalkanoate) (PHA-g-AA/t-CA)
were investigated in a study [341]. Structure, crystallization
behaviors and thermal properties of poly (3-hydroxybutyrate-co-
3-hydroxyvalerate)/organophilic montmorillonite (PHBV/OMMT)
nanocomposites were studied [342]. Structure–property relation-
ships for silica/poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)
(PHBHx) nanocomposites were investigated in a report [343].
In a study, preparation of biodegradable nanocomposites using
nanofibrillated cellulose (NFC) as the reinforcement and poly
(3-hydroxybutyrate-co-3-hydroxyvalerate, PHBV) as the polymer
matrix was reported [344]. The mechanical properties, biocom-
 A. Anjum et al. / International Journal of Biological Macromolecules 89 (2016) 161–174
patibility and biodegradability of composite materials made
from chestnut shell fibre (CSF) and poly (hydroxyalkanoate)
(PHA), as well as CSF and glycidyl methacrylategrafted PHA
(PHA-g-GMA), were evaluated [345]. Poly (3-hydroxybutyrate-co-
3-hydroxyvalerate) based nanocomposites containing different
amounts of organophilic montmorillonite were prepared using the
solution intercalation method. Thermal stability, tensile properties
and nanomechanical material properties was also investigated by
the nanoindentation technique [346]. Poly (3-hydroxybutyrate-
co-3-hydroxyvalerate) (PHBV)/cellulose nanowhisker (CNW)
composite with unidirectional CNW orientation were pre-
pared. CNW in poly(3-hydroxybutyrate-co-3 hydroxyvalerate)
(PHBV) matrix was aligned using an external electric field and
the morphology and mechanical properties of the resultant
anisotropic composites were studied [347]. Thermophysical
properties and biodegradation behavior of green composites
made from PHB and potato peel waste fermentation residue
(PPW-FR) fibers were investigated [348]. Nanocomposites of poly
(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx)/layered
silicates and PHBHHx/expanded graphite were synthesized.
For the nanofillers used in the two nanocomposites, an exfo-
liated morphology with good dispersion at lower nanofiller
content was observed [349]. Synthesis of a shape memory
polymer nanocomposite derived from poly (3-hydroxyoctanoate-
co-3-hydroxyundecenoate) composited with nanofillers of
silsesquioxane (POSS) was reported [350]. A nanocomposite
membrane based on a PHB-functionalized multi-walled carbon
nanotubes/chitosan matrix was used to efficiently pervaporate a
mixture of water and 1,4-dioxane [351].
8. Concluding remarks
Due to harmful effects of petroleum based plastic materials
in the environment and increasing concern about oil shortage
and petroleum prices, this paper focuses on the importance of
biodegradable plastics. Biodegradable polymers, especially poly-
hydroxyalkanoates, have gained a lot of interest both in research
and industry. Although their manufacturing costs today are still
too high to compete with conventional, petroleum- based poly-
mers, which is the major drawback of PHAs. Research is continuing
for the production of PHAs by utilization of low cost substrates
using wild and mutant strains of microorganisms, which could
improve the economics of the process. PHAs production by using
agriculture feedstocks, industrial by-products, waste plant oils and
wastewater is also very attractive which can make their produc-
tion environment friendly. This review also summarizes the most
recent advances in PHAs production using microbial strains and
the substrates towards their production. It can also provide unique
source of information about the blends and composites of PHA to
the scientists and researchers who are looking for the ideal replace-
ment of traditional plastics. In Future polyhydroxyalkanoates will
certainly play an important role in the plastics market due to their
versatility and novel features.
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