Accumulation

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© 1999 by Biopolymers
Press Inc. 389
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0273-2289/99/77–79/0389/$12.75

Accumulation of Biopolymers
in Activated Sludge Biomass

HONG CHUA,*,1 PETER H. F. YU,2 AND CHEE K. MA1

1
Department of Civil and Structural Engineering
and 2Department of Applied Biology and Chemical Technology,
The Hong Kong Polytechnic University, Kowloon,
Hung Hom, Hong Kong, China

Abstract
In this study, activated sludge bacteria from a conventional wastewater
treatment process were induced to accumulate polyhydroxyalkanoates
(PHAs) under different carbon-nitrogen (C:N) ratios. As the C:N ratio
increased from 20 to 140, specific polymer yield increased to a maximum of
0.38 g of polymer/g of dry cell mass while specific growth yield decreased.
The highest overall polymer production yield of 0.11 g of polymer/g of car-
bonaceous substrate consumed was achieved using a C:N ratio of 100. More-
over, the composition of polymer accumulated was dependent on the valeric
acid content in the feed. Copolymer poly(3-hydroxybutyrate-co-3-hydroxy-
valerate) [P(3HB-co-3HV)] was produced in the presence of valeric acid.
The 3-hydroxyvalerate (3HV) mole fraction in the copolymer was linearly
related to valeric content in the feed, which reached a maximum of 54% when
valeric acid was used as sole carbon source. When the 3HV U in the polymer
increased from 0–54 mol%, the melting temperature decreased from 178° to
99°C. Thus, the composition, and hence the mechanical properties, of the
copolymer produced from activated sludge can be controlled by adjusting
the mole fraction of valeric acid in the feed medium.

Index Entries: Activated sludge; carbon-nitrogen ratio; wastewater

treatment; butyric-valeric acid ratio; poly(3-hydroxybutyrate-co-3-
hydroxyvalerate).

Introduction
A wide variety of industrial and consumer plastic products have been
increasingly regarded as a source of solid waste-management problems
because of their nondegradable properties. In Hong Kong, 14 weight per-
cents of the 10,000 tons of municipal solid wastes produced each day are
plastic packaging materials and disposable products. Although plastics
*Author to whom all correspondence and reprint requests should be addressed.

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390 Chua, Yu, and Ma
usage and plastics-waste generation is forecast to increase at 15%/yr over
the next decade (1,2), there has been considerable interest in developing
environmentally-friendly materials to substitute for conventional plastics
(3,4). Polyhydroxyalkanoates (PHAs) have been recognized as better
candidates for biodegradable plastics owing to their similar material prop-
erties to conventional plastics and their complete biodegradability.
Approximately 300 different isolated microorganisms synthesize and
accumulate PHAs as carbon and energy storage materials under the condi-
tion of limiting nutrients in the presence of excess carbon source.
Among the various members of PHAs, only three kinds of PHAs, poly(3-
hydroxybutyrate) [P(3HB)], poly(3-hydroxybutyrate-co-3-hydroxyvalerate)
[P(3HB-Co-3HV)], and poly(3-hydroxyhexanoate-co-3-hydroxyoctanoate)
[P(3HHx-co-3HO)], have been produced to a relatively high concentration
with high productivity (5).
Although, PHAs have advantages over the conventional plastics,
widespread application of PHAs is hampered by high production cost. The
current cost of PHAs is ~10 times higher than that of conventional plastics
(3). Substantial reduction in production cost is necessary before widespread
applications in packaging and disposable products are possible. In the last
several years, fermentation strategies for the production of PHAs with high
productivity have been relatively well developed to bring down the overall
cost. Recently, much effort has been devoted to isolating bacterial strains
that can produce PHAs from cheap carbon sources.
Recently, many recombinant strains for economical production of
PHA have been developed by cloning the PHA synthase genes from many
microorganisms, including Alcaligenes eutrophus, Ectothiorhodospira shaposh-
nikovii, Pseudomonas aeruginosa, and Pseudomonas mendocina (5). Recombi-
nant Escherichia coli has been studied most extensively. The accumulation
of P(3HB) in recombinant Escherichia coli could reach 80–90% of dry cell
weight at the end of cultivation. However, the problem of the high oxygen
demand during the high cell-density culture of recombinant E. coli needs to
be solved to allow this process to be an economical alternative.
Owing to high production costs, a number of researchers have investi-
gated the use of cheap carbon sources as fermentation substrates. Methanol
is one of the cheap carbon sources available. However, it was demonstrated
that a long time was required for a methylotroph to produce large amounts
of P(3HB), resulting in low productivity (6). Moreover, it was suggested
that the high production cost for P(3HB) could be significantly decreased
by using photosynthetic bacteria with hydrogen and carbon monoxide as
inexpensive fermentation substrates (7). However, high capital costs
resulted owing to safety problems in using a flammable gas substrate. Many
researchers have demonstrated that high PHA production cost can be pos-
sibly lowered by using unpurified, low-cost organic wastes from agricul-
ture and food processing. Azotobacter vinelandii UWD, a new bacterial strain
for PHA production, was reported to produce PHA without nutrient limi-
tation and appears to be the best strain for PHA production from unpurified
substrate (8–10).
Applied Biochemistry and Biotechnology Vol. 77–79, 1999