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Jolanta PAJK, Micha ZIEMSKI, Boena NOWAK Faculty of Biology and Environmental

Protection, University of Silesia, Katowice


Please cited as: CHEMIK 2010, 64, 7-8, 523-530

Introduction
Vinyl polymers are derived from vinyl monomers containing double
bonds C=C. The main chain of polymer is made from carbon atoms
containing single bonds, to which other substituents can be added such
as chlorine, fluorine or hydroxyl group. Some polyloefins, polyvinyl
acetates, polyacrylates and others [1, 2] belong to vinyl polymers. Due
to their high molecular mass and considerable hydrophobic properties, they are classified as stable polymers (e.g. biodegradation time of
polyethylene is assumed to be ca. 300 years [3]).
In the environment, synthetic polymers undergo degradation influenced by abiotic agents and microorganisms [4]. Biodegradation
of materials is impeded as microorganisms have not formed suitable
mechanisms of their degradation. Nowadays, environmental pollution
caused by vinyl polymers originated, inter alia, from packaging poses
a serious problem. Therefore, a great pressure is put under the acquaintance with biodegradation pathways of polymers for which this
process is possible [5].
Poly(vinyl alcohol) is acknowledged to be one of few vinyl polymers that can have higher biodegradation rate. This is possible owing
to the presence of hydroxyl groups which condition hydrophilic nature
of this material [6].

dation (13% after 21-day period of incubation) was observed under


aerobic conditions in liquid cultures inoculated by sludge taken from
municipal wastewater. However, biodegradation degree of PVA performed in the presence of sludge taken from wastewater generated
in a paper-mill reached values comparable with the ones for cellulose.
The authors suggest that this can be caused by high selective pressure resulting from the presence of significant quantities of poly(vinyl
alcohol) in wastewater from paper factories. This pressure allows microorganisms present in such wastewater to gain abilities required for
degradation of this polymer [8].
The studies on PVA biodegradation under anaerobic conditions
were performed with the application of microorganisms from river
sludge and municipal wastewater. Foil samples with average (14 kDa)
and low (2.2 kDa) molecular mass were analysed. The important impact of polymer molecular mass on its degradation degree by microorganisms was reported. Microorganisms from river sludge caused a
75% decrement of mass of polymer with lower molecular mass, and
a 50-60% decrement of mass for samples characterized with higher
molecular mass in comparable incubation periods. Slightly lower biodegradation degrees of the polymer were observed for microorganisms originating from municipal wastewater [14].

Material biodegradation in various environments

Mechanisms of polymer biodegradation

Worldwide research into poly(vinyl alcohol) degradation in the


environment was initiated for its application in pulp and paper as well
as textiles industry which has resulted in generating significant quantities of wastewater containing PVA [7]. Poly(vinyl alcohol) degradation
was studied in, e.g. compost, soil, water, under aerobic and anaerobic
conditions.
Experiments conducted by Chellini et al. in compost from solid
municipal waste [8] demonstrated that biodegradation degree of
PVA foil containing 12% of acetate groups was 7% during 48 days. In
other studies [9], the same polymer was incubated for 300 days in the
presence of compost extract as the source of microorganisms. It was
shown that its 25% underwent biodegradation, whereas PVA with 2%
content of acetate group was degraded only in 15%.
The incubation of PVA foil in soil conducted by Chellini et al. [8]
demonstrated that 8-9% of this material undergoes biodegradation
within 74 days, and the level of this process is not affected by such
factors as polymer concentration or its physical state. Similar results
were obtained by Solaro et al. [10] during 120 days of PVA degradation
with 88% hydrolysis degree.
The

studies on the susceptibility of PVA


without acetate groups to degradation caused by soil microorganisms
were carried out by Sawada [11]. For this purpose, foil samples were
dug in 18 places representative of various types of soil and ambient
conditions. After two years of samples incubation, in spite of high
microbiological activity of soil, no traces of material colonization by
microorganisms was found and only slight decrement of foil mass (less
than 10%) was observed. Contrary to PVA, other polymers such as
PHB/HV or PCL were intensively degraded under the same conditioning conditions [12].
Also, the studies into degradation of poly(vinyl alcohol) in aquatic
environment are conducted. The

considerable degree of material


biodegradation was shown only in the presence of previously acclimatized microorganisms [13]. Relatively low level of PVA biodegra-

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Phytopathogenic fungi Fusarium lini were the first known microorganisms capable of PVA degradation. They synthesised an exoenzyme,
previously known as dehydratase, which action resulted in the release of carbon dioxide and water [15]. First microorganisms capable
of degrading this polymer as the only source of carbon were isolated
from soil samples and identified as bacterial strains called Pseudomonas
[16]. During further studies also other bacteria, such as Alcaligenes
and Bacillus, were isolated from environments polluted with PVA [17].
They are able to degrade this material as well.
The studies conducted by Watanabe et al. [18]

and Morita and


Watanabe [19] demonstrated that Pseudomonas strain releases two
exoenzymes which degrade poly(vinyl alcohol). They were identified
on the basis of hydrogen peroxide production during the reaction and
their ability to degrade some low molecular mass secondary alcohols
as secondary alcohol oxidase [EC 1.3.18 SAO] and -diketone hydrolase [EC 3.7.1.7 BDH]. SAO is highly specific to 1,3 hydroxyl
groups and catalyses oxidation of polymer molecules to adequate
-diketones. Then, they are attacked by a specific -diketone hydrolase which requires -diketone chain of length at least equal to 6 carbon
atoms for its complete activity.
This reaction results in products terminated with carboxyl and
methyl groups. Picture1 presents the biodegradation mechanism
through SAO and BDH [17].
Sakai et al. [20] proposed a biodegradation mechanism of
poly(vinyl alcohol) containing acetate groups by Pseudomonas vesicularis PD strain. It is based on the activity of SAO and BDH exoenzymes.
However, it was suggested that their degradation products containing
acetate groups can be uptaken and assimilated by bacterial cells.
Specific esterase present both in cytoplasm and cytoplasmic membrane turned out to be able to hydrolyse acetate groups. Its highest
activity was observed for poly(vinyl alcohol) molecules with low molecular mass and high content of acetate groups. Deacetylation cata-

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Poly(vinyl alcohol) biodegradable vinyl material

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Fig. 1. Pathway of PVA biodegradation with part of secondary alcohol
oxidase (SAO) and -diketone hydrolase (BDH) [17 altered]

lysed by esterase leads to the formation of such products as acetic


acid or hydroxy fatty acids. Then, they are metabolised within Krebs
cycle and -oxidation Picture 2 presents a biodegradation mechanism
of poly(vinyl alcohol) containing acetate groups.

Fig. 2. Biodegradation mechanism of poly(vinyl alcohol) containing


acetate esters [20 altered]

Matsumura et al. [21] were investigating the pathway of poly(vinyl


alcohol) degradation by specific dehydrogenase of PVA observed in
Alcaligenes faecalis KK314 strain isolated from river water samples.
This enzyme catalysis the formation of -hydroxyketone groups along
polymer chains, without their further oxidation to -diketones. They
also isolated an enzyme responsible for dissection of formed -hydroxyketones. Comparison of molecular masses with gel filtration profiles of both enzymes demonstrated that examined proteins are the
only enzyme which has two active places: dehydrogenase and aldolase,
whereby CaCl2 and Pyrroloquinoline Quinone (PQQ) as coenzyme
are required to conduct dehydrogenation reaction. Picture 3 presents
a mechanism of poly(vinyl alcohol) biodegradation by PVA dehydrogenase with the presence of Alcaligenes faecalis KK314.
Hatanaka et al. [22] isolated another PVA dehydrogenase depending on PQQ from Pseudomonas sp. 113P3 culture. The lack of enzyme
activity in supernatant indicated that PVA had to be uptaken inside the

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Fig. 3. Mechanism of PVA biodegradation by Alcaligenes faecalis


KK314:
PVADH

PVA dehydrogenase, PQQ - Pyrroloquinoline Quinone [21]

cell, and there it was degraded by specific intracellular dehydrogenase.


This enzyme catalysed the formation of -diketone groups along polymer chains, which were hydrolysed to low molecular weight PVA by
a specific hydrolase.
Shimao et al. [23] were the first who observed poly(vinyl alcohol) biodegradation in mixed cultures of some strains capable of PVA
assimilation. All symbiotic cultures showed the presence of a strain
producing an enzyme which degrades PVA and a strain which provides
an essential cofactor [24]. The first cannot grow in axenic culture, but
after adding supernatant of second strain culture into this culture, it
acquires the ability to grow.
A pair of symbionts was made of, inter alia, strains of Pseudomonas
putida VM15A and Pseudomonas sp. VM15C. P. putida VM15A is a strain
which provides a cofactor, namely PQQ, whereas Pseudomonas sp.
VM15C strain has two constitutive enzymes combined with cytopla
smic membrane which degrade polymer. PVA oxidase belongs to such
enzymes. Its activity, contrary to second enzyme - PVA dehydrogenase, is not affected by Pyrroloquinoline Quinone provided by P. putida
VM15A. Moreover, cytochrome reduction occurs during poly(vinyl alcohol) oxidation. This could indicate that polymer oxidation is related
to a cell electron transport chain. It was suggested that PVA oxidase
can be the subunit of oligomeric enzyme, to which PVA dehydrogenase
belongs. PQQ can participate in the transfer of electrons from subunit
oxidising PVA to other electron acceptor than oxygen as PVA dehydrogenase depending on PQQ does not consume oxygen during its activity. This can also explain the importance of PVA dehydrogenase as the
primary electron acceptor during PVA metabolism under anaerobic
conditions [25]. Picture 4 presents the proposed mechanism of PVA
biodegradation in symbiotic culture of microorganisms.
From phylogenetic point of view, poly(vinyl alcohol) and other synthetic polymers appeared in the environment at relatively late stage
of organisms evolution. Therefore, the majority of microorganisms
are not able to consume PVA as the only source of carbon. However,
it is possible that enzymes degrading PVA, such as PVA dehydrogenase Pseudomonas sp. VM15C, have appeared due to relatively recent
evolutionary processes. PVA dehydrogenase seems to be functionally
unique because its function considerably differs from functions of other
quinoprotein dehydrogenases.

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PVA application

Fig. 4. Mechanism of PVA biodegradation in symbiotic culture of


microorganisms [21]

Mori et al. [26] were investigating the capacity for poly(vinyl alcohol) degradation in mixed culture. It was stated that Bacillus megaterium BX1 bacteria demonstrated limited growth and PVA degradation
in axenic culture, while significant growth in PVA assimilation was observed in the presence of other gram-positive bacteria. These bacteria
did not degrade PVA which implied that poly(vinyl alcohol) degradation
was the result of co-metabolism of both strains, however, it was not
related to PQQ release.
There are few reports on PVA biodegradation by fungi.
Basidium fungi Phanerochaete chrysosporium, commonly present
in the environment, was the organism present during the majority of
experiments. It is capable of degrading naturally occurring lignin, and
also such xenobiotics as polycyclic hydrocarbons (PAHs) or synthetic
polymers, such as polyethylene or nylon [27-29].According to Mejas
experiments [30], one of its lignolytic enzymes - lignin peroxidase (LiP)
degrades PVA by forming carbonyl groups and double bonds along a
polymer chain.
Biodegradation mechanism was proposed, in which epoxide is
formed at the first stage.
Then, double bonds were formed by elimination of water molecules. Formed free-radical intermediate product
with produced benzaldehyde can be decomposed in next stages. Picture 5 presents the mechanism of PVA biodegradation by Phanerochaete chrysosporium.

Fig. 5. Path of PVA biodegradation by Phanerochaete chrysosporium [7]

Larking et al. [31] were investigating biodegradation with the part


of Pycnoporus cinnabarinus which decays hard-degradable industrial
dyes. They stated that chemical oxidation of PVA or the presence of
co-metabolite glucose, accelerates material degradation. Laccase another lignolytic enzyme, was the only enzyme involved in this process.
Experiments conducted by Nishikawa and Hasegawa resulted
in identification of Saccharomyces, Lipomyces and Rhodotorula yeasts
which degrade and assimilate poly(vinyl alcohol). For PVA containing
12% of acetate groups, this process takes place regardless of acetic
acid and its salts or esters presence in the solution. PVA is also biodegradable in the presence of Endomyces, Zygosaccharomyces, Pichia

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Nowadays, poly(vinyl alcohol) is the most commonly produced


polymer soluble in water [32].
Poly(vinyl alcohol) foils are used in agriculture to deliver fertilizers,
pesticides, herbicides and fungicides, as well as to coat seeds. Such
prepared seeds are ready for germination only under adequate temperature and humidity conditions [33].
For environmental protection, poly(vinyl alcohol) can be used as
a carrier of microorganisms. Gel manufactured by Kuraray company,
composed mainly of PVA, has a porous, reticulate structure that can
trap and carry microorganisms. A single gel bead with a diameter of
4mm can hold up to 1 billion microorganisms. PVA gel is used as an
alternative to the activated sludge method in the treatment of wastewater [34].
For its biocompatibility, intoxicity and lack of carcinogenic properties as well as due to excellent chemical properties, PVA is applied in
modern technologies for use in medicine and pharmacy [35, 36]. The
systems of controlled release of drugs are produced from this material. They can have the form of soft contact lenses, active dressing or
tablets.
It is also used to manufacture surgery threads, implants, scaffolds
for cells culture and artificial organs. Due to its hydrophicility, PVA is
most commonly used in combination with other polymers, such as
poly(lactic acid) (PLA), poly(glycolic acid) (PGA) or poly(-caprolactone) (PCL) [3638].
We can distinguish four types of systems of controlled release of
active substance: controlled bulging, polymer dissolution, crystals dissolution and modulated. The system of controlled bulging is based on
drug release caused by bulging of crosslinked polymers affected by
water. The quantity of released active substance is controlled by the
size of bulged material. Specified doses of drug are released in the
system of polymer controlled dissolution due to gradual dissolution of
PVA. Then, this material is excreted by an organism what eliminates
the need to remove polymer implants. The system of controlled dissolution of crystals is based on the change of polymer phase from crystalline to amorphous phase. No necessity to use toxic cross-linkers is its
advantage. In the modulated system, the action of external stimulus,
such as put electric field, causes the release of a therapeutic substance.
Also, self-modulated systems are possible. They are activated after the
action of physiological or environmental stimulus in the surroundings
of polymer [39].
For example, painkillers [36], antibiotics [40], hormones [41], anticancer drugs [38], vitamins [42] and other substances can be delivered
through the systems of controlled release made from PVA.
In tissue engineering, PVA is applied as scaffolds for tissues rege
neration, particularly for cartilaginous tissue regeneration. The rege
neration process of these tissues is ceased at some moment because
of more difficult access to oxygen and nutritive substances. This can be
prevented by forming new blood vessels in a place of tissue regeneration. Matrices made from PVA are used for this purpose. They contain gelatin beads with basic Fibroblast Growth Factor (bFGF). After
subcutaneous implantation to the target place, this factor is gradually
released, leading to the growth of new blood vessels.
Also, the research into using poly(vinyl alcohol) for obtaining hybrid internal organs to replace damaged ones has been in progress.
The prototype of hybrid pancreas has been constructed from live animal cells and a membrane protecting these cells against destruction by
a receiver immune system. This membrane, made from PVA, is biocompatible with the ambient environment, not capsulated by tissues
and is permeable for oxygen, glucose and insulin [39, 43].

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and Nadsonia yeasts, but only if acetic acid derivatives appear as cosubstrates. Besides these species, biodegradation of poly(vinyl alcohol)
was not observed in axenic culture of yeasts [7].

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PVA is also used in embolization procedure as the agent which


blocks blood flow in blood vessels [44, 45].

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Jolanta PAJK - Doctor of biological sciences, specializing in Biochemistry,


Faculty of Biology and Environmental Protection, University of Silesia, a senior
lecturer.
Micha ZIEMIEC - student, Department of Biology and Environmental Protection, University of Silesia.
Boena NOWAK - Doctor of biological sciences, specializing in Biochemistry, Faculty of Biology and Environmental Protection, Silesian University,
Assistant Professor.

SUNNY
CHEMISTRY
SUNNY CHEMISTRY - the initiative
of the Program Council of the monthly magazine CHEMIK and the Polish
Association of Chemical Engineers, is
supposed to change awareness and
formulate a new way of thinking about
chemistry in the society and science,
technique and economy.
This project aims at strengthening the
role of chemistry as most important
and first-rate in solving contemporary
problems, as well as in other fields of
science and technology.

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