BIOLOGICAL SYSTEM AS REACTOR

FOR THE PRODUCTION OF

BIODEGRADABLE THERMOPLASTICS,
POLYHYDROXYALKANOATES
AKHILESH KUMAR SINGH1 and NIRUPAMA MALLICK2
1

11.1
11.2
11.3

Introduction....................................................................................282
Biodegradable Polymers ................................................................283
Polyhydroxyalkanoates ..................................................................284
11.3.1 Chemical Structure...........................................................285
11.3.2 Types of PHAs..................................................................286
11.3.3 Potential Applications for PHAs ......................................287
11.3.4 PHA Biosynthetic Pathways.............................................288
11.3.5 Accumulation of PHA Co-Polymers ................................294
11.3.6 Commercial Drawback of Bacterial PHAs ......................298
11.3.6.1 Attempts for Low-Cost PHAs Production
in Transgenic Plants.........................................298

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11.3.6.2 Attempts for Low-Cost PHAs Production

in Cyanobacteria..............................................303
11.3.6.3 Attempts for Low-Cost PHAs Production
with Inexpensive Substrates ............................305
11.4 Conclusion and Future Perspectives ..............................................311
Keywords ..................................................................................................312
References.................................................................................................312

Plastics have unquestionably been the miracle materials and attained an

inimitable place in contemporary material technology. These are omnipresent in the present day society, appearing in manners, which range from ordinary to high-tech, from essential to completely lavish. The way in which
these materials are being utilized has molded our impression of plastics not
only due to wonder materials but also as necessary evils.
Plastics have become attention-grabbing materials during 1940s, and
since then these are substituting glass, wood and other construction materials, and even metals in many industrial, domestic and environmental applications (Lee et al., 1991; Cain, 1992; Poirier et al., 1995; Lee, 1996). These
extensive applications of plastics are not only owing to their promising
mechanical and thermal properties but also because of their stability, durability and low-cost.
of cellulose with nitric acid in the mid-19th century. Since then the number
ates, polysulfones, etc. Durability and resistance to degradation are desirable
characteristics when plastics are in use. However, they cause problems for
disposal when not in use. Globally, a wide variety of these polymers are
cant quantities of these materials are dumped into the ecosystem as industrial
cant role in several short-live applications for instance packaging and these
denote the foremost part of plastic wastes (Rivard et al., 1995; Witt et al.,
1997; Muller et al., 2001). The mammoth scale per capita utilization of these
materials to the extent of 60 and 80 kg, respectively in Europe and USA has

generated a prominent havoc to solid waste management programme (Kalia

et al., 2000). Several goods composed of plastics are ended up after their
useful life as discarded wastes, which accounting up to 20% by volume of
can remain for several years because these materials are highly resistant to
microbial attack owing to their high molecular mass, large number of aromatic rings, unusual bonds and/or halogen substitutions, once discarded in
nature (Alexander, 1981). On account of these reasons, some communities
are now susceptible to the effects of discarded conventional plastics on the
environment, together with detrimental impacts on wildlife and on the aesthetic qualities of cities and forests.

There have been increasing public and scientific interests over the past
three decades in relation to the utilization and development of environmentally friendly polymers (biodegradable polymers) as an ecologically useful
substitute to plastics, which must still preserve the preferred properties of
synthetic conventional plastics; therefore presenting a solution for the persisting serious problem of plastic waste materials. Biodegradability, which is
nothing more than catabolism measures the capacity of to be broken down,
especially into nontoxic products, through the action of microbes. Amongst
microorganisms, bacteria and fungi are the most important members in the
biodegradation process. The disintegration of materials furnishes them with
precursors for constituents of cell in addition to energy for energy-demanding
processes. With the likely exclusion of coral reefs and other analogous structures, the biological world actively degrades what it synthesizes. Therefore,
biodegradable materials are generally the product of life itself.
The exploration for such degradable polymers has focused for the introduction of three categories of degradable plastic materials: semi-biodegradable,
photodegradable, and completely biodegradable. Semi-biodegradable plastics include the starch-incorporated plastics where starch is introduced to
grasp together the small polyethylene fragments. The thought behind starchisms live in the soil will attack the starch and liberate fragments of polymer,
which can be degraded subsequently. Bacteria in fact attack the starch but
are turned off through the polyethylene fragments, which as a consequence

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remain non-degradable (Johnstone, 1990). Photodegradable plastics possess

light sensitive groups, which introduced as additives straightforwardly into
the polymer backbone. Ultraviolet radiation is able to disrupt their polymeric
structure rendering them open to further bacterial attack (Kalia et al., 2000).
third kinds of biodegradable plastics are quite new and promising owing to
its real exploitation by bacteria. Included biodegradable plastics are polylactides (PLA), polyhydroxyalkanoates (PHAs), polysaccharides, aliphatic
polyesters, co-polymers and/ or blends of the above. Of all these, the microbially-synthesized PHAs offer ample potential.

PHAs are a group macromolecules accumulated as water insoluble storage

compounds in the cytoplasm of microbial cells. They have gained much concern over conventional plastics due to their biodegradability and geneses
from renewable resources. Intracellular degradation of endogenous storage
compounds, for example, PHAs involve their breakdown when supply of
limiting nutrient is reestablished. In addition, a vast group of PHAs degrading microorganisms occur in nature and is capable to degrade PHAs by utilizing secreted particular PHA hydrolases and PHA depolymerases, and the
degradation products can be used as carbon and energy source (Madison
and Huisman, 1999). The actions of these enzymes can differ and depend on
the monomer units of the polymer, the dimensions of the sample, its physical form, for example, crystalline or amorphous and prominently, the conditions of environment. The degradation rate with respect to a portion of
poly-3-hydroxybutyrate (PHB) is normally on the order of a few months
in case of anaerobic sewage to years in seawater (Madison and Huisman,
1999). Similarly, co-polymers incorporating 4-hydroxybutyrate (4HB)
monomers reported to degrade more rapidly as compared to poly-3-hydroxybutyrate (PHB) or co-polymer comprised of 3-hydroxybutyrate and
3-hydroxyvalerate [P(3HB-co-3HV)] (Williamson and Wilkinson, 1958).
Apart from biodegradability, another important feature associated with
PHAs is their synthesis, which is based on renewable resources, such as agricultural feedstocks resulting from CO2 and water. After the transformation
of agricultural feedstocks into biodegradable PHAs, the breakdown products
are once again CO2 and water. Therefore, while for certain applications the
biodegradability is crucial, PHAs gain comprehensive consideration as they

are based on renewable resources instead of on non-renewable shrinking

fossil fuel stocks (Williams and Peoples, 1996).

The several diverse PHAs, which have been documented so far, are mainly
linear, head-to-tail polymers made up of 3-hydroxy fatty acid monomers
(Madison and Huisman, 1999). In these polyesters, the carboxyl group
of one monomer establishes an ester bond with the hydroxyl group of the
adjoining monomer being represented by the general chemical structure as
exhibited in Figure 11.1, where X can arrive up to 30,000 and R-pendant
group comprises H-atom or a large varieties of C-chains.
For example, when
n=1

R = hydrogen

methyl
ethyl

n=3

Poly(3-hydroxybutyrtate)

Poly(3-hydroxyvalerate)

propyl

Poly(3-hydroxyhexanoate)

nonyl

Poly(3-hydroxydodecanoate)

R = hydrogen

Poly(5-hydroxyvalerate)

pentyl
n=2

Poly(3-hydroxypropionate)

R = hydrogen

Poly(3-hydroxyoctanoate)
Poly(4-hydroxybutyarte)

So far, in all PHAs, which have been characterized the hydroxyl

replaced carbon atom is of R con uration, except in certain special circumstances where there is no chirality. At the identical C-3 or position an
alkyl group that can differ from methyl to tridecyl, is placed. Nevertheless,
this alkyl side chain is not essentially saturated; unsaturated, aromatic,
epoxidized, halogenated and branched monomers have been reported too
(Lageveen et al., 1988; Abe et al., 1990; Doi and Abe, 1990; Fritzsche et al.,
1990a, b, c; Kim et al., 1991, 1992; Choi and Yoon, 1994; Hazer et al.,

General structure of polyhydroxyalkanoates.

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1994; Curley et al., 1996; Song and Yoon, 1996). In the side chains of
PHAs, substituents can be altered chemically such as by cross-linking of
unsaturated bonds (de Koning et al., 1994; Gagnon et al., 1994a, b). These
differences in the length and composition of the side chains and the capacity to alter their reactive substituents are the foundation for the diversity
of the PHA polymer family and their huge array of potential applications
(Madison and Huisman, 1999).

Polymers of 3-hydroxyalkanoates (3HAs) have been established in a wide

range of prokaryotes and in several eukaryotic plants and animal cells.
However, only prokaryotes are proficient of synthesizing high molecular
weight PHAs in the form of amorphous granules with insignificant osmotic
activity. To date, more than 300 species of microorganisms are identified to
build and accumulate PHAs (Lee et al., 1999).
PHAs can be assigned into three key categories based on the number of
carbons in the monomer units integrated into the polymer chain, for example, short-chain-length (SCL), medium-chain-length (MCL) and long-chainlength (LCL) PHAs composed of hydroxyacids with 35, 614 or more than
14 carbon atoms, respectively (Anderson and Dawes, 1990; Steinbuchel,
1992; Steinbuchel et al., 1992). Interestingly, it is not viable to produce
PHAs chemically; therefore, microorganisms are the merely potential source
for these polymers. PHAs are non-toxic, biocompatible, piezoelectric, thermoplastic and/or elastomeric, enantiomerically pure, insoluble in water and
exhibit a high degree of polymerization with molecular weights of up to
several million Da (Muller and Seebach, 1993; Hocking and Marchessault,
1994). Amongst the 150 diverse types of PHA monomer units known so
far, the homopolymer of 3-hydroxybutyrate (PHB) is common in different
taxonomic groups of prokaryotes (Vincenzini and De Philippis, 1999; Reddy
et al., 2003). The monomer composition of the PHAs is regulated by two
most important factors, for example, the type of microorganism used and the
type of substrate exploit to grow the microorganism (Valappil et al., 2007).
This generates an opportunity for synthesizing various types of PHAs with
a widespread range of properties (Reddy et al., 2003). Because of the steall in D() con

uration (Reddy et al., 2003). PHAs can be advocated for

various commercial applications, owing to resemblances of material properties with conventional plastics. The potential applications of the PHAs
vary depending on material properties governed by monomer composition
(Valappil et al., 2007). Poly-3-hydroxybutyrate (PHB), which is the common member of SCL-PHAs is quite rigid and crystalline, therefore making
it problematical to process and be commercially exploited (Park and Lee,
2004). In contrast, MCL-/LCL-PHAs are elastomeric materials depicting
poor tensile strength. Owing to these limitations, SCL-, MCL- and LCLPHAs as such not feasible for various industrial applications. However, integration of MCL- or LCL-monomer units into the PHB backbone transforms
the material properties of the polymer, making it appropriate for various
commercial application Matsusaki et al., 2000; Singh and Mallick, 2008,
2009a, b; Singh et al., 2013).

The foremost attractive features of these polymers are their biodegradability together with hydrophobicity; even if economical biodegradable plastics
are commercially available, merely PHAs have a good moisture resistance,
so that their use in application for which these characteristics are required,
for example, packaging of liquids and food materials, papers coating, etc.
can be envisaged. The practicable applications of PHAs as plastics, related
to their more important properties, and their real commercial applications
(Vincenzini and Philippis, 1999) are in the areas of: (i) Agriculture: PHAs
could be utilized for controlled release of pesticides, plant growth regulators, herbicides and fertilizers owing to their biodegradability coupled with
retarding properties; (ii) Medical: PHAs biocompatibility and biodegradability together with piezoelectric properties lead to potential application
as absorbable sutures, bone plates, film around bone fracture, surgical pins
and staples; (iii) Pharmaceutical: Owing to their biodegradability and biocompatibility, PHAs could be employed as drug carrier and retarded drug
release; (iv) Disposables: PHAs could be used as sole structural materials
such as razors, tray for foods, utensils, etc. because of their biodegradability
with good mechanical properties; (v) Packaging: PHAs could be exploited
for manufacturing bottles and film for food packaging owing to their biodegradability, good mechanical properties, moisture resistance, hydrophobic
and low oxygen permeability.

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Bacterial species producing PHAs have been fundamentally subdivided

into three groups (Nomura et al., 2005). One group together with the bacterium Alcaligenes eutrophus, now termed as Cupriavidus necator, yields
SCL-PHA, whereas a second group in conjunction with a number of
Pseudomonads, produces MCL-PHA (Steinbuchel, 1991). The third group,
including the bacteria Aeromonas hydrophila and Pseudomonas sp. 613
accumulates SCL-MCL-PHA co-polymer composed of both SCL and
MCL monomer units. In contrast, sludge-isolated Pseudomonas aeruginosa MTCC 7925 has been known so far to synthesize a novel SCL-LCLPHA co-polymer composed of LCL 3-hydroxyhexadecanoate (3HHD) and
3-hydroxyoctadecanoate (3HOD) units with SCL 3-hydroxybutyrate (3HB)
and 3-hydroxyvalerate (3HV), for example, P(3HB-co-3HV-co-3HHD-co3HOD) as constituents of PHAs (Singh and Mallick, 2008, 2009a, b; Singh
et al., 2013). This partition between SCL-PHA, MCL-PHA, SCL-MCLPHA and LCL-MCL-PHA co-polymer is predominantly determined by the
substrate specificity of the PHA synthase liable for the polymerization of the
substrate R-3-hydroxyacyl-CoA to form PHAs.
PHB is the most common and comprehensively characterized PHA present in bacteria. It is synthesized as a carbon and/or energy storage material in
a number of microorganisms generally under limiting nutritional conditions
for instance N and P stresses and/or in presence of excess carbon sources
(Steinbuchel, 1991; Byrom, 1994; Liebergesel et al., 1994; Yu, 2001).
PHB biosynthesis and degradation are best studied in a few bacterial spetablished in 1973 for Azotobacter
beijerinckii and Cupriavidus necator. In these microorganisms along with
many other bacteria when they are grown on carbohydrates or on acetylCoA producing carbon sources, the biosynthetic pathway for the production
of PHB starts from acetyl-CoA and involves three consecutive, enzymemediated reactions, for example, the action of 3-ketothiolase, acetoacetylCoA reductase and PHA synthase (Oeding and Schlegel, 1973; Senior and
the reversible condensation of two acetyl-CoA moieties to form acetoacetylCoA. Acetoacetyl-CoA reductase consequently reduces acetoacetyl-CoA to
R-()-3-hydroxybutyryl-CoA, which is then polymerized by the action of a
PHA synthase to form PHB (Figure
of this pathway has been detected in Rhodospirillum rubrum, which needs

PHB biosynthesis in Alcaligenes eutrophus (Cupriavidus necator).

Enzymes: (1) 3-Ketothiolase, (2) NADPH-linked acetoacetyl-CoA reductase, (3) PHA
synthase, (4) PHA depolymerase, (5) 3-hydroxybutyrate dehydrogenase, and (6) AcetoacetylCoA synthase (Vincenzini and De Philippis, 1999).

the additional action of two enoyl-CoA hydratases to synthesize R-()-3hydroxybutyryl-CoA (Moskowitz and Merrick, 1969), but without altering
the basic rules.
On the basis of types of PHAs that are produced by Cupriavidus necator one general observation is that the integrated monomers always consist
of only 35 carbons. The nature and ratio of these monomers are neversupplemented to the growth media (Doi et al., 1988; Steinbuchel, 1991;
Steinbuchel et al., 1993). It has been revealed that supplementation of propionic acid or valeric acid to the growth media with glucose results into production of P(3HB-co-3HV) co-polymer, the biosynthetic pathway of which
is presented in Figure 11.3.
The PHA biosynthesis in Pseudomonas oleovorans and in many
Pseudomonads including Pseudomonas aeruginosa belonging to the
rRNA homology group I involves the biosynthetic route that includes the
cyclic oxidation along with thiolytic cleavage of fatty acids, for example, 3-hydroxyacyl-CoA and intermediates of the -oxidation pathways

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Biosynthetic pathway of PHB and P(3HB-co-3HV) in Alcaligenes eutrophus

(Cupriavidus necator) (modified from Valentin and Steinbuchel, 1995; Suriyamongkol
et al., 2007).

(Doi, 1990; Punrattanasin, 2001; Singh and Mallick, 2009a; Singh et al.,
2013). These microorganisms synthesize MCL-PHAs and rarely LCL-PHAs
from alkanes, alkanoates or alcohols. Contrary to Cupriavidus necator, these
organisms normally do not synthesize PHAs comprising SCL monomers
(SCL-PHAs). The -oxidation of fatty acids could supply the foremost suban enzymatic cascade of reactions where fatty acids are incorporated into the
cell (Figure 11.4): they are activated to CoA thioesters, reduced to 2-transenoyl-CoA, and catalyzed by acyl-CoA dehydrogenase with FAD as a cofactor. These intermediates are transformed to S-(+)-3-hydroxyacyl-CoA and
catalyzed by enoyl-CoA hydratase. The compound so generated is oxidized
to 3-ketoacyl-CoA with the assistance of NAD dependent 3-ketoacyl-CoA
dehydrogenase. Lastly, acetyl-CoA is cleaved from 3-ketoacyl-CoA and
two carbons lesser fatty acid is synthesized. By reduction of 3-ketoacylCoA, a reaction catalyzed by a ketoacyl-CoA reductase, intermediate R-()3-hydroxyacyl-CoA is produced. As the PHA synthase accepts merely the
R-()-3-hydroxyacyl-CoA and the bacterial -oxidation of fatty acids produces merely the S-(+)-3-hydroxyacyl-CoA, bacteria must have enzymes
R-()-3-hydroxyacyl-CoA. One such enzyme is
a 3-hydroxyacyl-CoA epimerase, mediating the reversible conversion of
the S and R isomers of 3-hydroxyacyl-CoA. Thus, S-(+)-3-hydroxyacylCoA epimerizes to R-()-3-hydroxyacyl-CoA through 3-hydroxyacyl-CoA
epimerase and the enoyl-CoA is converted to R-()-3-hydroxyacyl-CoA by

Biosynthetic pathway of MCL-PHAs and LCL-PHAs in Pseudomonas

oleovorans and Pseudomonas aeruginosa, respectively using oxidation cycle. Enzymes:
1: Acyl-CoA synthase, 2: Acyl-CoA dehydrogenase, 3: Enoyl-CoA hydratase, 4: NAD
dependent 3-hydroxyacyl-CoA dehydrogenase, 5: Ketoacyl-CoA thiolase (reductase),
6: 3-Hydroxyacyl-CoA epimerase, 7: Enoyl-CoA hydratase, 8: Ketoacyl-CoA reductase and
9: PHA polymerase (Lagaveen et al., 1988; Huisman et al., 1989; Singh and Mallick, 2009a;
Singh et al., 2013).

merase for the production of either MCL-PHAs or LCL-PHAs (Lageveen

et al., 1988; Kraak et al., 1997; Singh and Mallick, 2009a).
In P. aeruginosa nevertheless, PHA is produced from acetyl-CoA
through fatty acid biosynthetic pathway (Huijberts et al., 1995; Steinbuchel,
1996; Singh and Mallick, 2008, 2009b). Various Pseudomonads belonging
to the rRNA homology group I too accumulate MCL-PHAs by this pathway. However, Pseudomonas aeruginosa MTCC 7925 produce a novel
co-polymer composed of LCL 3HA units of C16 (3-hydroxyhexadecanoate,
3HHD) and C18 (3-hydroxyoctadecanoate, 3HOD) along with SCL 3HA

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units of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). In these

microorganisms the pathway operated is known as the P. aeruginosa PHA
biosynthetic pathway. In most cases, the building blocks of PHAs are incorporated into the polymer only if the cells are cultivated on carbon source
whose carbon skeleton is related to the structure of the constituent monomer
unit (Huisman et al., 1989). In some cases, of course, the PHA monomers do
not have similarity with the carbon sources; this was referred as the capability of the organism to synthesis PHA from unrelated substrates (Anderson
and Dawes 1990). Steinbuchel (1996) and Singh and Mallick (2008) reported
that MCL- and LCL-PHAs synthesized by P. aeruginosa PHA biosynthetic
pathway are from unrelated substrates, for example, glucose, ethanol, gluacids, which then yield precursors for PHA polymerase to produce either
MCL- or LCL-PHA polymer (Figure 11.5). The fatty acid synthesis pathway
exhibits different biosynthetic pathways according to the growth substrates
to acetyl-CoA and then carboxylated to malonyl-CoA. A malonyl transacylase links the malonyl-CoA to an acyl carrier protein (ACP), liberating
coenzyme A. Malonyl-ACP is further converted to 3-ketoacyl-ACP, R-()is then reacted to a malonyl-ACP to produce a new 3-ketoacyl-ACP molecule. In this pathway, R-()-3-hydroxyacyl-ACP is a presumed precursor
for the production of PHA. As the fatty acid biosynthesis intermediate is
in the form of R-()-3-hydroxyacyl-ACP, an additional biosynthetic step is
required to transform it into the R-()-3-hydroxyacyl-CoA form. An enzyme
way to PHA biosynthesis. Nevertheless, P. oleovorans does not synthesize
MCL-PHAs from these unrelated substrates. The difference between these
two related organisms is that P. oleovorans degrades fatty acids to form acylCoA,
P. aeruginosa
produces acyl-ACP (acyl carrier protein), a precursor substrate for fatty acid
production from unrelated substrates resulting in either MCL- or LCL-PHA
(Figure 11.5).
As stated above, the majority of bacteria produce either SCL-, MCL- or
rarely LCL-PHAs but not generally SCL-MCL or SCL-LCL co-polymers
because of the mutual exclusivity of the SCL-, MCL- and LCL-PHA synthase enzymes (Kato et al., 1996; Ashby et al., 2002; Singh et al., 2013).

Biosynthetic pathway of MCL-PHAs and LCL-PHAs in Pseudomonas

aeruginosa using fatty acid synthesis pathway. Enzymes: 1: Ketoacyl-ACP synthase,
2: Ketoacyl-ACP reductase, 3: 3-Hydroxyacyl-ACP dehydratase, 4: Enoyl-ACP reductase,
5: 3-Hydroxyacyl-ACP-CoA transacylase and 6: PHA polymerase (Timm and Steinbuchel,
1990; Steinbuchel, 1996; Singh and Mallick, 2008; Singh and Mallick, 2009b).

Biosynthesis of PHAs, in which SCL and MCL or SCL and LCL monomers are covalently linked in the same polyester molecules, for example,
the real co-polymer of SCL-MCL or SCL-LCL needs the existence of a
PHA synthase showing a substrate range combining those of SCL-PHA and
MCL-PHA or SCL-PHA and LCL-PHA synthases. Interestingly, in some
cases presence of SCL- and MCL-PHA, where the monomer units not connected in the same polyester molecule has also been detected (Steinbuchel
and Wiese, 1992; Kato et al., 1996; Ashby et al., 2002). Simultaneous presence of a SCL-PHA synthase and a MCL-PHA synthase is the reason for

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Industrial Biotechnology: Sustainable Production and Bioresource Utilization

formation of such mixture of SCL- and MCL-PHA rather than the real SCLMCL co-polymer.

As indicated in Table 11.1, some bacterial species such as Aeromonas

caviae, Rhodococcus sp. NCIMB 40126, Bacillus sp. and some species of
Pseudomonas (P. fluorescens, Pseudomonas sp. A33, Pseudomonas sp.
613, P. marginalis DSM50276 and P. mendocina DSM50017) are found
to synthesize the real co-polymer of SCL-MCL-PHAs with a maximum carbon skeleton up to C14 (Haywood et al., 1991; Caballero et al., 1995; Doi
et al., 1995; Lee et al., 1995a; Kato et al., 1996; Tajima et al., 2003; Nomura
et al., 2005; Li et al., 2011; Gao et al., 2012; Phithakrotchanakoon et al.,
2013). However, Pseudomonas aeruginosa MTCC 7925 produces a novel
SCL-LCL-PHA co-polymer with incorporation of a maximum carbon skeleton up to C18. Thus, for synthesis of the co-polymer of SCL-MCL-PHAs
and SCL-LCL-PHAs, Pseudomonads are believed to be of the appropriate
candidates, due to their inherent properties of synthesizing MCL-PHAs and
LCL-PHAs (Huisman et al., 1989; Singh and Mallick, 2008, 2009a, b; Singh
et al., 2013).
Bacterial system appropriate for biosynthesis of SCL-MCL-PHA and
SCL-LCL-PHA co-polymer is of current research concern owing to their
interesting material properties (Chen et al., 2001b; Lee and Park, 2002;
Singh and Mallick, 2008, 2009a, b; Singh et al., 2013; Phithakrotchanakoon
et al., 2013). The SCL-MCL-PHA and SCL-LCL-PHA co-polymers are
found to have superior material properties as compared to those of PHAs
comprising of SCL or MCL or LCL monomers only (Doi et al., 1995; Singh
and Mallick, 2008, 2009a, b; Singh et al., 2013). It has been revealed that
SCL-MCL-PHA co-polymers with a high mol percentage of SCL monomers
along with a low mol fraction of MCL and SCL-LCL-PHA co-polymers with
high mol percentage of SCL monomers in conjugation with low mol fraction
of LCL have properties comparable to polypropylene (PP) and polyethylene
(PE) (Matsusaki et al., 2000; Abe and Doi, 2002; Noda et al., 2004; Singh
and Mallick, 2008, 2009a, b; Singh et al., 2013). Co-polymer of P(3HBco-3HHx) containing 1017 mol% of 3HHx retains higher elongation to
break as compared to P(3HB-co-3HV) co-polymer containing 20 mol% of
3HV, commercialized under the trade name of BIOPOL (Lee et al., 2000b).