From Obtaining To Degradation of PHB: Material Properties. Part I

Ingeniería y Ciencia
ISSN:1794-9165 | ISSN-e: 2256-4314

ing. cienc., vol. 13, no. 26, pp. 269–298, julio-diciembre. 2017.
http://www.eafit.edu.co/ingciencia
This article is licensed under a Creative Commons Attribution 4.0 by
From Obtaining to Degradation of PHB:

Material Properties. Part I
Antonio José dos Santos1 , Luiz Veriano Oliveira Dalla Valentina2 ,
Andrey Alayo Hidalgo Schulz3 and Marcia Adriana Tomaz Duarte4
Received: 31-03-2017 | Accepted: 08-08-2017 | Online: 14-11-2017
PACS: 81.05.Lg; 82.35.Lr; 82.35.Pq; 87.15.rp
doi:10.17230/ingciencia.13.26.10
Abstract
This paper presents a review of the chemical, physical and morphologi-
cal characteristics, as well as the existing applications and mechanisms for
the production of poly (3-hydroxybutyrate). This biopolymer, which is
obtained from renewable sources, degrades when exposed in biologically
active environments and is biocompatible, that is, it is not rejected by the
human body in health applications. However, in spite of presenting simi-
lar properties with some conventional plastics, the PHB exhibits fragile
behavior and thermal instability when processed. The literature proposes
the use of blends, the development of copolymers or the insertion of addi-
tives in an attempt to improve the mechanical and thermal properties of
PHB.
Key words: Biopolymers; polyhydroxyalkanoates (PHA);
Polyhydroxybutyrate (PHB).
1
Centro Universitario Sociesc - UNISOCIESC, [email protected], https://
orcid.org/0000-0002-2539-2499, Joinville, Brazil.
2
Universidade do Estado de Santa Catarina - UDESC, [email protected], https://
3
Centro Universitario Sociesc - UNISOCIESC, [email protected], https://orcid.org/
0000-0002-4525-2418, Joinville, Brazil.
4
Centro Universitario Sociesc - UNISOCIESC, [email protected], https://
Universidad EAFIT 269|

From Obtaining to Degradation of PHB: Material Properties. Part I
De la obtención a la degradación de PHB:

Propiedades del material. Parte I
Resumen
Este artículo presenta una revisión de las características químicas, físicas
y morfológicas, así como las aplicaciones y mecanismos existentes para la
producción de poli(3-hidroxibutirato). Este biopolímero, que se obtiene a
partir de fuentes renovables, se degrada cuando se expone en ambientes
biológicamente activos y es biocompatible, es decir, no es rechazado por
el cuerpo humano en aplicaciones de salud. Sin embargo, a pesar de pre-
sentar propiedades similares con algunos plásticos convencionales, el PHB
exhibe comportamiento frágil e inestabilidad térmica cuando se procesa.
La literatura propone el uso de mezclas, el desarrollo de copolímeros o la
inserción de aditivos en un intento por mejorar las propiedades mecánicas
y térmicas del PHB.
Palabras clave: Biopolímeros; degradación de PHB; polihidroxialcanoa-
to (PHA);
polihidroxibutirato (PHB).
1 Introduction
Throughout history, man has withdrawn from the nature materials and
products for their survival and life quality. Other products used by man
are synthesized from materials also arising from nature. An example of this
are the polymers [1]. These materials are being produced in ever greater
proportions and have replaced other materials in certain areas [2],[3]. In
1960 the polymers corresponded to less than 0.5% of urban solid waste
generated in the USA and in 2009 this figure already represented 12% [4].
According to Abiplast [5], in Brazil, 13.5% of the total waste generated
arise from the polymeric materials segment.
The majority of synthetic polymers is produced from fossil fuel, a non-
renewable natural resource, and is intended for the packaging industry [5].
Due to its fast disposability, combined with its difficult degradation, plastic
packages are increasing and accumulating in the environment [6]. This
fact is causing a growing concern with regard to environmental problems
and the depletion of natural resources. In addition, this situation has
led to reflection on more sustainable alternatives for production, use and
|270 Ingeniería y Ciencia

Antonio J. Santos, Luiz V. O. Dalla Valentina, Andrey A. H. Schulz and Marcia A. T.
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disposal of polymers with the objective of minimizing the environmental

damage. Among the alternatives to minimize this problem are recycling
and incineration [7].
Another alternative proposed for the management of these residues are
the biodegradable materials, being these more accessible to microbial assi-
milation, since for most synthetic polymers the degradation is a process
that happens slowly and its speed depends on environmental conditions and
the composition of polymers [8]. The use of biodegradable polymers is an
environmentally correct alternative that can replace the not biodegradable
polymers, in certain applications [8].
In this scenario, the poly(3-hydroxybutyrate) PHB, is a polymer that
has been causing growing interest in scientific and industrial world, be-
cause besides having thermoplastic properties and physical and mechanical
characteristics similar to conventional plastics, is a biodegradable material,
biocompatible and can be obtained from renewable sources [9],[10],[11].
However, the cost of biopolymers production, as the PHB, is high and this
makes them more expensive than the synthetic polymers [1],[12]. In Brazil,
the PHB is produced by a bacterium through fermentation of sugar cane
[12], which makes its cost of production economically interesting, since the
sugarcane is planted in several regions [13].
The purpose of this paper is to make a literature review on the PHB,
as well as on the various existing mechanisms for their production.
2 Poly(3-hydroxybutyrate)(PHB)
The poly (hydroxyalkanoate) (PHAs) constitute a group of thermoplastic

polyesters, which possess properties similar to some conventional plastics
from petrochemical industry [9],[14], featuring thermoplastic characteristics
and water resistance in relation to the synthetic polymers [15]. The PHAs
are biodegradable and synthesized by bacterial way, from renewable raw
materials, such as, pure sugar, fatty acid or fermented liquor from waste
[16]. Among the PHAs the poly(3-hydroxybutyrate), P(3HB) stands out.
The poly(3-hydroxybutyrate) PHB is one of the most studied polymers
occurring more frequently when the PHAs, are accumulated by microor-
ganisms [17],[18],[19],[20].
ing.cienc., vol. 13, no. 26, pp. 269-298, julio-diciembre. 2017. 271|
The literature assigns the discovery of PHB Maurice Lemoigne, who

isolated it and characterized it in 1926 [21]. However, the date of the
discovery differs among the authors of the area. Marchessault [22] mentions
the period between 1923 to 1925, once that Vroman and Tighzert [23] state
that this polymer began to be produced from 1925.
The PHB is an aliphatic polyester [24] with linear polymer chain [18],
which presents thermoplastic properties [25]. This material is nontoxic
[9], optically active [26] and when pure, has a yellowish coloration [6],[27],
presents barrier to permeability of water and gas [6] and stability to ultra-
violet radiation [28], and can be processed by extrusion, injection, blowing
and thermoforming [29].
One of the most mentioned characteristics in the literature about the
PHB is its biodegradability. This only occurs, however, when the material
is exposed to environments biologically active [30] (soils, sea water or fresh
water and aerobic and anaerobic composting), which are environments in
which the material is in contact with degrading microorganisms [31]. It can
also be mentioned the activated sludge as another environment proper to
the degradation of PHB [32] and the sanitary landfill, in which the material
may be discarded without impact to the environment [6].
Another important characteristic of PHB is the biocompatibility, which
can be understood as a satisfactory interaction between certain substance
and the physiological environment in which is inserted. In other words,
is the term used when the material is not rejected by the body and when
the assimilation generates non-toxic waste. The PHB is biocompatible
for two reasons: it is present in human bloodstream, in the form of PHB
of low molar mass, and due to its degradation product be the acid 3-
hydroxybutyric, a common metabolite in living beings [33].
The experiments conducted in the study of HUDA et al. [10] feature the
PHB as a source of bioenergy, once that this material can be converted into
methane gas when biodegraded. This material is also considered chemically
recycable, being that the studies evidenced this recycling through experi-
ments performed with the PHB [19],[34],[35].

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2.1 PHB synthesis
The PHB is a material which also generates interest by presenting different

forms of being synthesized [6]. There are three routes of obtaining PHB,
being first by means of polymerization synthetic by ring opening [23]. This
route uses the β-butyrolactone as reagent [23]. In this process, zinc and alu-
minum can be used as catalysts [32] and the polymer obtained shall possess
chiral center in configuration in both R, as in S, being the last named PHB
racemic compound [36]. Another method of obtaining is through natural
plants and/or transgenic plants [11]. And the third, and more widespread,
strategy for the synthesis of this polymer is by means of bacterial fermen-
tation [1],[2],[29],[37], pure or mixed, commonly known as biosynthesis. In
this process the substrate is used as a carbon source [28]. It is important
to emphasize that the stereoregularity of the polymer depends on your
route of obtaining [23] and the stereospecificity of the enzyme responsible
for the polymerization of the monomer [38]. There are several species of
bacteria able to accumulate the PHB and this number exceeds 70 different
genera [39]. These bacteria can be found in a variety of environments, in
soil, water, sewer microbiologically treated [40] and even of mangrove [15].
Some of the most known species of PHB bacteria are: Ralstonia eutropha
[41] (nowadays Cupriavidus necator [42]), Escherichia coli [9], Burkholde-
ria cepacia and Zobellela denitrificans [15], Bacillus megaterium and Pseu-
domonas oleovorans [26], Protomonas extorquens, Protomonas oleovorans,
Alcaligenes latus and Azobacter vinelandii [43]. These bacteria can be di-
vided into two groups according to the composition of the culture medium
necessary for the synthesis of PHB. The first group needs, during its expo-
nential growth phase, of an environment that is deficient in nutrients (N,
Mg, S, P, K or O) and that has an excess of carbonic reserve. This situation
induces the interruption of microbial cellular growth and the consequent
consumption of reserve of carbon for production of PHB. The other group
does not require nutritional deficiency in culture medium [26].
When metabolized by micro-organisms, the substrate is stored interce-
llularly as source reserve of energy [25] e carbon [44]. This buildup occurs
in the cytoplasm of cell in the form of granules, with diameters ranging
from 0.2 µm to 0.5 µm [2].
The space occupied by granules can reach rates between 70 % to 90 % of
the dry mass of the cell [38]. The proposed structure for these granules is a
hydrophobic core composed by PHB involved by a phospholipid monolayer.
This arranges enzymes responsible for synthesis and degradation of PHB,
called respectively of PHA synthase and PHA depolymerase [45]. The
Figure 1 illustrates what was previously described.
Figure 1: (a) Granules of PHB accumulated in the cell of the bacterium Azobac-
ter chroococcum [2]; (b) Structure of the granule of PHB [45].
After this first stage of accumulation of PHB inside the bacterial cell
(Figure 1a), the polymer is extracted and purified until reaching the shape
of the final product dry and solid, when it will be ready to be processed
[46]. A characteristic of the polymerization of PHB that attracts world
attention is that this can be operated on mild processing conditions without
the minimal environmental impact [47].
The mechanism of synthesis of PHB can be described in three stages
as follows [35]: (a) consists in the dimming of the acetyl coenzyme A
in acetoacetyl-CoA, catalyzed by enzyme β-ketothiolase (PhaA); (b) the
PHA-reductase enzyme (PhaB) catalyzes the hydrogenation of acetoacetyl-
CoA, forming the monomer (R)-3-hydroxy-butyryl-CoA; (c) occurs the
polymerization of (R)-3-hydroxy-butyryl-CoA, catalyzed by enzyme PHB-
polymerase (PhaC), forming the PHB.

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The initiation of polymerization of (R)-3-hydroxy-butyryl-CoA compre-

hends the connection of two molecules of the same monomer the two thiols
groups, belonging to the amino acids cysteine that constitute the active
site of PHB-polymerase enzyme. Further it occurs the condensation of two
monomers that leave one of two free thiol groups. The spread happens
when a third monomer joins the thiol free group, resulting in another con-
densation and so forth, generating the growth of the polymer chain of PHB
[2]. Figure 2 demonstrates the proposed mechanism.
Figure 2: The mechanism of synthesis of PHB by Ralstonia eutropha [2].
In order to increase productivity, Coats, Watson and Brink-Man [48],

for example, developed a study involving a microbial consortia cultured on
fermented dairy manure, which under certain control conditions showed a
significant reduction in PHB production costs. Similarly, Wu, Chen and
Chen [49] showed that the combination of a new cell growth pattern with
a morphologically altered E. coli reduced the time required for the produc-
tion of PHAs, increasing the efficiency of the process of obtaining these
biopolymers.
2.2 Chemical characteristics
As a member of the PHAs, the poly(3-hydroxybutyrate) is characterized

by having a methyl (CH3 ) as alkyl replacing group [23], which confers to
the material hydrophobic characteristics [6],[29]. In addition, this polymer
is composed of monomers of 3-hydroxybutyrate that have 4 to 5 carbon
atoms, which is considered a short chain lenght [26]. This monomer that
contains the β-hydroxybutyric acid [18], which has an alcoholic group (-OH)
and a carboxylic acid (-COOH) [24]. Another characteristic of the chemical
structure of PHB is the presence of a chromophoric carbonyl group [28].
This is illustrated in Figure 3.
Figure 3: Chemical structure of PHB [50].
2.3 Physical properties
The distribution of molar mass of a polymer is the measure of the dis-

tribution of their individual molar masses around the average molecular
mass [26]. In the case of PHB, this value is quite high, in range of 10,000
to 3,000,000 g/mol [26]. This range of variation depends on the producer
micro-organism, of carbon source and concentration, of the duration of the
fermentation, the growth rate and of the polymer purifications conditions
[26]. The critical molar mass of this material (indicative value to which
any value below this means marked loss on the mechanical properties of
the polymer) is estimated at 160,000 g/mol [51].
An important characteristic of PHB is their crystallinity, because it de-
fines the physical and mechanical properties of the polymer [52]. The crys-
talline state of a material is described as being one that displays a structure
set, therefore a polymer that has this characteristic is called semicrystalline.

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And it is so named because its long polymer chain will never crystallize in
whole as it presents a crystalline phase and another amorphous [52]. The
crystalline phase of PHB depends on the regularity of its structure, which
in turn depends on the route that this was synthesized. O isotactic PHB
[47] has chiral carbon in absolute configuration R [26],[38] and is obtained
by means of bacterial fermentation [23], while the syndiotactic PHB is syn-
thesized from monomers with setting R and S track synthetic route [53].
In this way, the first presents higher crystallinity than the second. In this
context, the PHB presents a regular structure [54] and this will allow high
crystallinity [55]. Among the studies made about this material, not every-
one agrees with the range of crystallinity presented by PHB. Vroman and
Tighzert [23] claim that this crystallinity is above 50 %, while Sharma and
Ray [56] say to be above 60 %. In both studies mentioned previously a
maximum value of crystallinity was not stipulated, as what has been pro-
posed by Sadi et al. [28] who declared the crystalline range of PHB as
being between 50 % to 80 %. It is observed that this band of crystallinity
varies widely among researchers 55 % to 80 % [57], or 60 % to 70 % [58],
or 60 % to 80 % [18],[59] or 60 % to 90 % [52],[53].
2.4 Crystalline structure
In the crystalline phase of PHB, the molecules are arranged in an organized

and repetitive way, where the smallest repetitive unit is called a unitary
cell. This is composed by 4 monomeric units in a orthorhombic system,
with the following dimensions: a = 5.76 angstrom, b = 12.30 angstrom and
c = 5.96 angstrom [60]. The molecules have helical conformation in L (in-
dicating swing to the left), where each complete revolution is made by two
monomeric units. And present as basic forces of the model conformational,
interactions of Van der Walls between the oxygen of carbonyl and methyl
groups [26]. This structure may be visualized in Figure 4.
Figure 4: (a) Helical structure proposed for the molecule of PHB (the bar the
axis of the polymer chain; (b) Projections of single cell of PHB in plans cb and
ab [61].
2.5 Morphology of PHB
The PHB presents different structures which can be sequenced in accor-

dance with the increasing order of its dimensions: lamellae, fibrils and
spherulites. The lamellae are crystals in tape format composed by packag-
ing in parallel of molecules of PHB, organized in a way that the axis of the
polymer chain (c axis) is perpendicular to the faces of tape [62]. The set
of lamellae packaged in parallel is called fibrils, in which the unitary cell
of PHB is arranged so that the axis "a" is oriented parallel to the radius
of the spherulite [63]. The union of anchoring fibrils in a central point and
irradiated in all directions, with spherical shape, is called spherulites [64],
that in the case of PHB are few, but large [28]. Figure 5 exemplifies these
structures.

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Figure 5: (a) lamellae of extended chain; (b) lamellae of folded chain [62]; (c)
fibrils - packaging of lamellae [65] and (d) spherulite - showing fibrils and lamellae
[66].
Figure 6: PHB spherulites seen with optic microscopy [67].
Through an optical microscope of polarized light, the structure of the

spherical spherulite can be displayed and you can still see a pattern of the
Maltese cross and concentric light and dark bands, which arise by rotation
of the axes "b" and "c" around the axis "a" of unitary cell [63]. Figure 6
illustrates this situation.
Barham et al. [52] present a study that relates the spacing increase
of bands and the radius of spherulites with the increase of temperature
of crystallization of PHB. In addition, the low rate of nucleation of PHB,
which generates few cores in the crystallization, makes large spherulites
[51]. The thickness of the lamella (lc) is equal to the length of the segment
of the polymer chain crystallized in lamella, for the PHB varies from 5
nm to 20 nm [62] and this variation depends on the molar mass, of crys-
tallization temperature and the cooling rate from the molten state [50],
[61],[62]. When the molar mass of PHB is less than 2 × 10−3 g.mol−1 , the
widths of the lamellae will be larger or closer to the length of the extended
chain, which represents that the molecules are fully extended (Figure 5a).
However, when the molar mass is greater than 2 × 10−3 g.mol−1 lamellar
thicknesses will be lower than the length of the extended chain, which indi-
cates that the molecules bowed to adjust to the lamellar crystals (Fig 5b)
[62]. The amorphous phase of PHB is located in the interlamellar spaces
and intrafibrilar of the spherulites [66]. The composition of this stage is
of folds of chains, by segments of not crystallized string in lamellae and
fraction of chain whose end is crystallized in adjacent lamellae.
2.6 Temperatures and transitions
It is interesting to know the values of some specific temperatures of poly-

mers to learn how the material behaves in a temperature range, and in
this way to know what the possible processabilities and applications of the
same are. It is the case of the glass transition temperature (Tg), the melting
temperature (Tm), temperature that initiates the reduction of molar mass,
temperature of initiation of degradation and main temperature of degrada-
tion. Before starting to present the values of such temperatures, it is valid
to elucidate the significance of Tg e Tm. In a semicrystalline polymer as
the PHB, glass transition is the change of state from the glass state to the
liquid state, in its amorphous phase. Therefore, Tg is the one in which such
change happens. When temperatures are below the Tg, the segments of
the chain and pending groups have restricted mobility. Now when the tem-

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peratures are above the Tg, the molecules of the polymer feature disabled
translational and rotational degrees of freedom and vibrational [68].
The Tg depends on molecular parameters, such as, tacticity, molar
mass, degree of crystallinity, polydispersion, branches or pending groups
on the main chain [68], and experimental variables, such as, sample prepa-
ration, thermal history, rate of heating or cooling [69]. It is interesting to
clarify a little more about the dependence that Tg of PHB has in relation
to two of these factors: the molar mass and the degree of crystallinity.
Through studies observed that the Tg grows with the increase of molar
mass of PHB up to 2 × 10−3 g.mol−1 , when, from this level it remains
constant [62]. Now regarding the influence of the lamellar crystals in glass
transition of PHB, it was observed that the higher the degree of crys-
tallinity the higher the Tg. This is explained by the crystallization process
of material that causes the confinement of segments of molecules in the
interllamelar and interfibrilar spaces, which restrict translational mobility
of the segments. In view of this, it is necessary the heating of the material
to submit these segments to the glass transition [70]. Depending on the
author the Tg can have different values. Some authors suggest that the
glass transition temperature is approximately 5 ◦ C [6],[28],[57]. However,
values around 4 ◦ C to 7 ◦ C [56], or around 0 ◦ C to 5 ◦ C [71], or with higher
temperature oscillating, in the range -15 ◦ C to 9 ◦ C [55], can be found.
In its turn the melting temperature is that in which the crystalline re-
gions disappear because of the fusion of the crystallites. Due to occurring
in the crystalline phase, the Tm only makes sense if applied in semicrys-
talline polymers as the PHB, for example. And it is in this temperature
that the system power surpasses the secondary intermolecular forces of
polymer chain of the crystalline phase. As a result it occurs the destruc-
tion of regular structure of packaging and the change of status from rubber
for the state viscous, also called the molten state [68]. The fusion of the
crystalline phase of poly(3-hydroxybutyrate) is an irreversible process that
can be evidenced through three ways [62],[72]:
The variation of the parameters Tm (melting temperature) and ∆H◦ m
(enthalpy of fusion) with the heating rate submitted to the material;
Super cooling ∆t = T◦ m - Tc, indicated by the displacement of crys-
tallization temperature (Tc) in relation to the melting temperature in the
balance (T◦ m). When a process is reversible, both temperatures are iden-
tical;
Lamellae formed by folded chains. When a process is reversible poly-
meric chains are completely unfolded in the lamellae. The thickness of the
lamellae, the molar mass and the crystallization temperature are factors
that directly influence in the melting temperature of the PHB. Tm in-
creases with thicknesses of larger lamellae, with the elevation of the molar
masses up to 2 × 10−3 g.mol−1 , from this value the melting temperature
varies little in a given level and with the growth of crystallization tempe-
rature, when there is the formation of thicker lamellae [62],[72]. The same
situation of temperature variation reported in the literature, but with other
values, happens with the melting temperature. Some authors [10] mention
that this value is 173 ◦ C, but other values can be found: 175 ◦ C [6],[73]
or 180 ◦ C [23],[60]. There are studies that mention ranges of variation for
this temperature: 175 ◦ C to 180 ◦ C [56], or 170 ◦ C to 180 ◦ C [53], or 160
◦ C to 180 ◦ C [18], ranges which cover all the values mentioned previously.
When submitted to temperatures above 170 ◦ C, the PHB begins to lose

molar mass [6], which leads some authors affirm that the temperature of
degradation is close to this value [71]. Now, in agreement with the studies
of Mousavioun et al. [11], the temperature that initiates the degradation
of PHB is 212 ◦ C and the main temperature of degradation is 260 ◦ C.
The melting temperature in balance (T◦ m) can be conceptualized as
the melting temperature of the lamellae with infinite thickness, which in-
dicates that the polymer chains are fully extended in crystals and melting
temperatures and crystallization are the same [52]. T◦ m can vary between
179 ◦ C to 186 ◦ C in accordance with the molar mass, being the first value
for a weighted average molar mass (Mw) of 20,000 g/mol and the second
value for molar mass of 200,000 g/mol. This oscillation corresponds to the
influence that the terminations of the chains exercise under the crystalliza-
tion of the polymer [74].
The enthalpy (∆H◦ m) is usually found through the integration of the
peak corresponding to the fusion process that is observed by DSC [52].
The ∆H◦ m crystallized PHB from molten state is in function of molar
mass and the history of the thermal material. However, the molar mass
only influences in the enthalpy of fusion until values equal to 2 × 10−3

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g.mol−1 , for values higher than the enthalpy is practically independent of

molar mass [62]. For the PHB the value found for the enthalpy of fusion is
of 146 J/g [25]. Some of the properties described and mentioned previously,
in addition to others, can be found in Table 1.
Table 1: PHB properties
Property Value Reference

Density (g/cm3 ) 1.23 Machado [57]
Molecular mass (x 105 ) 0.1 to 30 Sudesh [26]
Glass transition (◦ C) 5 Casarin et al. [6]
5 to 6 Rodrigues et al. [75]
4 to 7 Sharma [56]
0 to 5 Ayorinde [71]
-15 to 9 Srubar III et al. [55]
Melting (◦ C) 173 Mousavioun et al. [11]
175 Casarin et al. [6]
180 Santos et al. [59]
175 to 180 Sharma [56]
170 to 180 Jendrossek [53]
160 to 180 Alejandra et al. [18]
Melting at the balance (◦ C) 179 to 186 Barham et al. [52]
Enthalpy of fusion (J/g) 146 HO et al. [25]
2.7 Aging and mechanical characteristics
Soon after processing the PHB goes through slow changes in the physical
properties of both phases (amorphous and crystalline), causing the har-
dening and the weakening of the material. This process is known as the
aging of PHB and its nature comes from two phenomena, the secondary
crystallization and physical aging [76].
After being processed, the molten PHB begins to cool and thus it be-
gins its process of crystallization, which can be divided into three parts. In
the first stage, the material cools without the crystallization of its struc-
ture. Now in the second phase, the crystallization occurs in high rates,
characterized as autocatalytic or crystallization. Finally, in the last stage

of the process, the crystallization succeeds in low rates, reaching a stage of
crystallinity self-balance, also called secondary crystallization [64].
This last step of crystallization (secondary), crystallization occurs in the
amorphous phase of the polymer at ambient temperature during storage
of the product [23]. It is a phenomenon known by formation of imperfect
crystals in the spaces of interlamellar spherulites and by the restriction of
movement of the amorphous phase, which is unable to dissipate energy by
viscous flow [77].
Now the physical aging can be described as the molecular relaxation
that occurs due to residual mobility of polymeric chains of glass phase
at temperatures below the Tg. Thus, polymeric chains of glass phase are
capable of consuming changes conformational, which generates a reduction
of the free volume. Therefore, the smaller the free volume, the lower the
mobility of polymeric chains and that consequently brings in the hardening
of the material [78].
The consequences that both phenomena carry to the physical properties
of PHB are on the degree of crystallization, resistance to impact, Young’s
modulus, the tenacity and elongation at break [79]. The increase in the
crystallinity of PHB, over time, shows that secondary crystallization occu-
rred PHB and composites, reaching stability around 20 days with values of
crystallinity varying between 43% and 47%, independently of the load pres-
ence [80]. The Young’s modulus increases, indicating the rigidity of the ma-
terial. In turn, the impact resistance decreases, evidencing the weakening
of PHB. The value of the elongation of rupture also decreases considerably
after two or more weeks of aging [79].
As already explained, the aging exerts influence on some properties of
PHB as the fragility that according to Gutierrez-Wing et al. [19] decreases
with the reduction of the crystallinity, the rigidity [25] and hardness [6].
These characteristics grant to the PHB low mechanical properties and pre-
vent its ability from withstanding big impacts [11]. Table 2 presents some
values of mechanical properties of PHB.

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Table 2: Mechanical properties of PHB [81].
Property B189C-1/18BC-1 B189D-1 B1000

Tensile strength (MPa) 30 32 36
Elongation at break (%) 2.2 4.0 2.0
Young’s modulus (MPa) 2600 2200 3800
Impact resistance (J/m) 23 28 36
*Data provided by the manufacturer according to ASTM standards,
considering different formulations; impact resistance by Izod test.
The study of crystallization and melting behavior is basic to understand

and control of material properties and processing necessary to get them. An
experiment was developed in order to understand the effect of the thermal
cycles in the polymer phase transition suggesting that the crystallinity of
PHB can be controlled by the thermal cycles of heating and cooling to
which the material is subjected [82].
2.8 PHB and improvement measures
The PHB presents properties similar to some conventional plastics, as the

polypropylene [83]. Authors affirm that these characteristics are physi-
cal and mechanical [25]. PHB also resembles polyethylene on thermo-
mechanical properties [56] and with PLA in relation to mechanical proper-
ties [23]. Despite of this, a difference of PHB, besides its fragile behavior,
it is its thermal instability [29], being susceptible to thermal degradation
when processed [55]. This occurs because of their close processing window,
defined by the proximity between its melting and degradation tempera-
tures [28]. In addition, this material flows during processing [6]. Literature
proposes several approaches for the PHB aiming to improve its mechanical
behavior [25] and reduce its high cost [84]. These approaches are based on
the use of blends [11], obtaining copolymers [85] and insertion of additives
[29].
Ke et al. [21] discuss the physical interaction of the reactive polymer
blends and the formation of copolymers during the reactive polymer blen-
ding. They comment that the compatibilization can occur either by adding
compatibilizers and/or the induction of chemical or physico-chemical inter-

action during the blending process, for example, crosslinking the compo-
nents, chemically modifying the homopolymers, etc. These authors cite
that the introduction of intermolecular H-bonds to polymer blends is an
effective method to improve the miscibility between two components. In
this case, the crystallinity and melting temperature decrease for PHB or
PHBV, which is beneficial to modify the mechanical properties and en-
large their processing windows. Another approach to improving properties
of PHAs (linear polymers with minimal entanglement) is the formation of
branching/crosslinking copolymers via reactive polymer blending. Dicumyl
peroxide (DCP) possessing a relatively high hydrogen abstraction ability,
makes it ideal as a branching or cross-linking agent. The same authors
also discussed two other possible techniques to improve the performance of
blends: formation of graft copolymers via reactive polymer blending and
the formation of complex copolymers via reactive polymer blending.
Studies in the literature are found regarding the PHB/ PP blends
[28],[83], combination of PHB with aromatic polymers [50]. Mousavioun
et al. [11] points blends of PHB with poly (caprolactone) (PCL), poly
(vinylidene) fluoride (PVDF), poly (vinyl alcohol) (PVOH), poly (lactic
acid) (PLA), poly (vinyl acetate) (PVA), poly (vinyl phenol) (PVP), ce-
llulose ester, and furthermore, is based on its work on blend PHB/lignin.
Burlein [86] introduces in his work the blend PHB/LDPE. HO et al. [25]
still mentions blends with inorganic nanoparticles, as the magnetite with
size in the order of nanometers.
Bordes et al. [86] and Ke et al. [21] confirm the possibility of inser-
ting hydroxyvalerate units (HV) between polymeric chains. It is obtained
as a result of the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate)
(PHBV), which presents lower melting point, increased this way the pro-
cessing window, after the treatmentresearches of Wei, Mcdonald and STARK
[87],[88], PHB or PHBV were grafted with cellulose via reactive extrusion
insitu DCP as the initiator. Results on surface morphology obtained by
SEM revealed a better compatibility of the cellulose in the polymer matrix
(PHB or PHBV) in relation to the results obtained due to the modifica-
tion by means of blends. The grafting reduced the degree of crystallinity
and increased the tenacity and flexibility of the biocomposites. It has been
found that this simple reactive extrusion process extends the processing

Duarte
window for PHB-based biocomposites, which increases the range of possi-

ble applications of these biomaterials. Larsson, Markbo and Jannasch [89],
showed that the termal stability of the PHAs at the processing temperature
can be dramatically improved by simply washing the materials in a 1 mM
aqueous HCl solution. Hence, the thermal decomposition temperature in-
creased by up to 50 ◦ C after the treatment. Subsequently, treated PHB and
poly(3-HB-co-4-HB) were blended with different amounts of poly(butylene
adipate-co-terephthalate) by melt extrusion in order to further enhance
the processability and thermomechanical properties. After adding DCP
during the extrusion, the interfacial adhesion improved, and the dynamic
shear and tensile storage modulus increased with increasing content of the
peroxide.
According Garcia-Garcia et al. [90], the compatibility of a blend PHB
with PCL was improved by reactive extrusion with different DCP con-
tents, led to a remarkable increase in ductile properties such as elongation
at break and impact-absorbed energy. Addition of 1 wt % DCP also con-
tributes to lowering the degree of cristallinity of PHB. Wei and Mcdonald
[91], conducted a study blending PHB and PLLA, that were individually
cross-linked with DCP (0.25 - 1 wt %) by reactive melt processing. The
size of the polymer crystal spherulites, Tg, Tm and cristallinity were all de-
creased as a result of cross-linking. Cross-linked polymers were also shown
to be more thermally stable than linear polymers with broader molecular
weight distribution (MWD) and longer chain branching (LCB) and conse-
quente improvement in melt strength. These improvements in the proper-
ties of these materials increase your processing options, such as foaming
and thermoforming.
When comparing the copolymerization products to the blend it is po-
ssible to say that this is a much faster and easier way to obtain the desired
properties. Additionally, when making blends there is the possibility of
incorporating cheaper polymers in relation to PHB, reducing costs and im-
proving its mechanical properties [15],[44]. The introduction of additives,
such as, nucleating agents, plasticizers, lubricants, antioxidants and pho-
tostabilizers, is another approach. The mixture of PHB with nucleating
agents makes the average size of spherulites decrease preventing the for-
mation of large cracks and interspherulitic on the effect of weakening [55].
In addition, the plasticizers and lubricants in a blend have functions to
improve molecular mobility in the molten state, decrease the Tg, reduce
the termal degradation and improve the impact resistance of PHB.
2.9 Applications
Due to the characteristics of the biodegradability and biocompatibility pre-

viously submitted, the PHB presentes great potential for applications in
pharmacological area [92], environmental [9], packaging [11], veterinary and
industrial [17], agricultural [18] and biomedical [21],[25]. Some examples of
applications are presented in Table 3.
Table 3: Examples of applications of PHB.
Area Application
Biomedical Sutures, supports of tissue cultures for implants, sur-
gical implants, dressings, part of bones and replanted
veins, engineering of heart valves and pins.
Pharmacological Encapsulation of medicines for controlled release
Packaging Food packaging
Veterinary Encapsulation of veterinary medicinal products
Agricultural Encapsulation of fertilizers
Environmental Bags, bottles, disposable items, items of personal hy-
giene, films of involvement, degradable diapers and re-
mediation of areas affected by oil spills.
Industrial Recovery of oligomers and monomers for new use in
the synthesis of polymers
It can be mentioned, as environmental benefits generated by the use

of this polymer, the energy required in production processes, which is ob-
tained by burning of sugarcane bagasse; biomass from the extraction of the
polymer and effluent processes that can be applied such as fertilizers in the
planting of sugarcane; and the use of solvents, natural and biodegradable,
originated from the alcoholic fermentation, for purification of the polymer,
all without causing negative environmental impact [30]. Due to the high
cost of production of PHA (generally), its commercial exploitation is sig-
nificantly lower than expected The industrial scale production of PHA is

Duarte
limited by various factors, including the high cost of raw materials, low
productivity of microbial processes (i.e., 3-4 g/(l h) PHA), the high cost of
downstream processing and the separation of PHA from biomass cellular
[93].
3 Conclusions
Polymeric materials have been used more and more frequently in society
due to their mechanical resistance, lightness, chemical inertia and low cost
of production. For this reason, these materials eventually replace metals,
wood and glass in some applications. The accumulation of waste from the
disposal of plastic products produced with synthetic polymers is a problem,
considering the low speed of their degradation.
An ecological alternative prior to this situation is the production and
use of biopolymers such as PHB. These materials are therefore classified
because of the characteristic of degrade under the influence of environ-
mental factors. PHB is a hard and brittle thermoplastic polyester with a
high crystallinity index, is not soluble in water, has low thermal stability
and properties that resemble polypropylene. This polymer can be obtained
chemically by ring-opening polymerization and by biosynthesis. The latter
is the pathway for the production of PHB best known and is performed
by bacteria that synthesize and store the polymer in intracellular granules
within its own cytoplasm.
Its cost of production is still high compared to other synthetic polymers,
however, Brazil is one of the countries that has the lowest cost to produce
PHB due to the sugarcane cultivation conducted by the alcohol industry,
which is used as a substrate for its industrial production. This polymer has
applications in the biomedical, pharmacological, veterinary, agricultural,
environmental, industrial and mainly in the packaging sector. All these
applications are possible due to two very important characteristics of PHB:
biocompatibility and biodegradability.
In this review, we seek to show aspects related to the properties, the
process of obtaining and the possibilities of improving the performance of
the PHB. The second part of this review will discuss the different forms
of degradation suffered by the PHB, at the end of its life cycle, and the
necessary requirements for its occurrence.
Acknowledgements
The authors would like to acknowledge the financial support from CNPq
- National Council for Scientific and Technological Development (Brazil).
We would also like to thank the companies PHB Industrial S/A and Arkema
Química Ltda for supplying the polymeric materials used in this research.
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Ingeniería y Ciencia

ISSN:1794-9165 | ISSN-e: 2256-4314

From Obtaining to Degradation of PHB:

Received: 31-03-2017 | Accepted: 08-08-2017 | Online: 14-11-2017

PACS: 81.05.Lg; 82.35.Lr; 82.35.Pq; 87.15.rp

Universidad EAFIT 269|

De la obtención a la degradación de PHB:

|270 Ingeniería y Ciencia

disposal of polymers with the objective of minimizing the environmental

The poly (hydroxyalkanoate) (PHAs) constitute a group of thermoplastic

The literature assigns the discovery of PHB Maurice Lemoigne, who

|272 Ingeniería y Ciencia

2.1 PHB synthesis

The PHB is a material which also generates interest by presenting different

|274 Ingeniería y Ciencia

The initiation of polymerization of (R)-3-hydroxy-butyryl-CoA compre-

Figure 2: The mechanism of synthesis of PHB by Ralstonia eutropha [2].

In order to increase productivity, Coats, Watson and Brink-Man [48],

2.2 Chemical characteristics

As a member of the PHAs, the poly(3-hydroxybutyrate) is characterized

Figure 3: Chemical structure of PHB [50].

2.3 Physical properties

The distribution of molar mass of a polymer is the measure of the dis-

|276 Ingeniería y Ciencia

2.4 Crystalline structure

In the crystalline phase of PHB, the molecules are arranged in an organized

2.5 Morphology of PHB

The PHB presents different structures which can be sequenced in accor-

|278 Ingeniería y Ciencia

Figure 6: PHB spherulites seen with optic microscopy [67].

Through an optical microscope of polarized light, the structure of the

2.6 Temperatures and transitions

It is interesting to know the values of some specific temperatures of poly-

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When submitted to temperatures above 170 ◦ C, the PHB begins to lose

|282 Ingeniería y Ciencia

g.mol−1 , for values higher than the enthalpy is practically independent of

Table 1: PHB properties

Property Value Reference

2.7 Aging and mechanical characteristics

characterized as autocatalytic or crystallization. Finally, in the last stage

|284 Ingeniería y Ciencia

Table 2: Mechanical properties of PHB [81].

Property B189C-1/18BC-1 B189D-1 B1000

The study of crystallization and melting behavior is basic to understand

2.8 PHB and improvement measures

The PHB presents properties similar to some conventional plastics, as the

compatibilizers and/or the induction of chemical or physico-chemical inter-

|286 Ingeniería y Ciencia

window for PHB-based biocomposites, which increases the range of possi-

Due to the characteristics of the biodegradability and biocompatibility pre-

It can be mentioned, as environmental benefits generated by the use

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|290 Ingeniería y Ciencia

[10] S. Huda, “Anaerobic digestion of polyhydroxybutyrate accumulated in excess

[21] Y. Ke, X. Y. Zhang, S. Ramakrishna, L. M. He, and G. Wu, “Reactive blends

|292 Ingeniería y Ciencia

chain length polyhydroxyalkanoates,” Polymer International, vol. 59, no. 5,