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ELSEVIER
Intemational Biodeterioration & Biodegradation (1995)
249-268 Copyright 0 1995 Elsevier Science Limited Printed in Great
Britain. All rights reserved
0964-8305/95/$9SO+.OO 0964-8305(95)00033-X
Bioremediation of Alpechin
A. Ramos-Cormenzana, M. Monteoliva-Sanchez & M. J. Lopez
Department of Microbiology, Faculty of Pharmacy, University of
Granada, Granada, Spain
ABSTRACT
Olive oil extraction produces large amounts of residues. These
olive-mill wastes are known as alpechin. Alpechin-polluted waste
waters are resistant to degradation. They are regarded as a severe
environmental problem because of their high organic content,
largely simple phenolic compounds that are both antimicrobial and
phytotoxic.
During the past years, most of the investigations related to
alpechin treatment focused on the microbial degradation of the
alpechin polyphenols.
However, the actual perspectives are based on both the
bioremediation of alpechin and on the modification of the
technology used in olive-oil extrac- tion. In this article we will
describe the most relevant biotreatment systems that can be used
for the recycling of alpechin wastes. We will also discuss
for each system their prospects for future uses. The various
systems we will discuss include the following: (I) Bioremediation
for use as fertiliser or soil conditioner. (2) The utilisation as a
medium for grown edible mushrooms. (3) The application as a growth
medium for algae in open basins. (4) Biopolymeric substances
production from alpechin, focusing on poly- sacharide and
biodegradable plastics production. (5) The use as a bioe- nergetic
source (or for biogas production). (6) The employment of alpechin
as a source of biopharmaceuticals.
INTRODUCTION
Olive oil extraction produces a large volume of solid and liquid
residues, the olive-mill waste waters are known as alpechin (Fig.
1). The world production is widely distributed, but the two most
important olive oil producers are Spain and Italy (Fig. 2).
249
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250 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
WATER 1.250
WATER 1.375Kg
Fig. 1. Olive by-products.
Olive oil waste effluents are major pollutants and constitute a
great problem in olive tree cultivation areas. In several European
countries including Italy and Spain, it is illegal to dispose of
the wastes directly into waterways, and they have to be stored in
closed or open ponds.
Closed ponds generate large amounts of sludge, which has to be
disposed. These types of closed ponds for alpechin storage are
sources of contamination for surface and ground waters.
In the case of open ponds, the water is evaporated by sunlight
all year long, and a sludge is created, which must be disposed of.
This method needs large areas of arable land and is generally
insufficient in all respects.
Open and closed ponds generate many volatile malodorous
compounds. Both systems are very costly due to high labour and
maintenance expen-
SPAIN 41%
PORTUGAL 2%
Fig. 2. European Community olive oil production.
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Bioremediation of alpechin 251
ses. Moreover, in these systems, the water and the valuable
substances contained within the wastes which represent potential
financial incomes are lost.
During past years, efforts were made to seek a viable solution
to solve the problem and the overall objectives of most research
programs focused on strategies that would help increase the
commercial potential of alpe- chin. The first investigations deal
with the microbial degradation of the alpechin polyphenols. In
order to prevent industrial pollution, new highly sophisticated
treatment plants are built in polluted geographical areas. The
schemes presented on Figs 3 and 4 illustrate strategies that are
proposed to reduce alpechin generated pollution. Some people
believe that the two-phase olive oil production system shown on
Fig. 4 will solve the pollution problem. However, this system
generates a new waste product, called Alpeorujo in Spain which will
have to be treated. In addition, the two-phase process is very
costly.
The problem concerning the alpechin is derived from its
composition. Many references can be found on this subject-Moreno et
al. (1990), Ramos-Cormenzana (1986), Fiestas Ros de Ursinos (1958)
and others. The fundamental composition is essentially water,
organic substances (organic acids, phenols - which are considered
responsible for the toxic properties - miscellaneous organic
compounds) and minerals. However, the alpechin composition is
highly variable, when considering each individual component, mainly
because of the fact that the alpechin
u ELIMINATE ) 4 w
Fig. 3. Solutions proposed for the alpechin environmental
problem.
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252 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
OLIVE OIL SYSTFM PRODUCTION QLIVE OIL TWO PHASES SYSTFM PRODU-
-k
d OLIVE
io,,, &
OIL
Fig. 4. Olive oil extraction technology.
is a natural product, processed from a raw material and subject
to varying conditions which are difficult to control. In this sense
the forthcoming new technologies should take this into account. The
olive oil waste water is resistant to degradation and represents a
severe environmental problem because of its high organic content,
largely simple phenolic compounds (Gonzalez et al., 1990) that are
both anti- microbial and phytotoxic. Therefore, new alternatives
are actually needed for their disposition.
However, not all the effects of alpechin are adverse and there
are some important beneficial effects. For example, olive oil waste
waters also contain valuable substances which could be recovered.
Some of these can stimulate the soil nitrogen-fixing bacteria
population or the production of extracellular slime which
contributes to the stability of soil aggregates and hence the tilth
of the soil and its water-retention capacity. Other compo- nents of
these wastes can suppress soil-borne fungal foot pathogens, such as
Pythium, Phytophthora spp.
Moreover, these waste products could be used as a substrate in
biotechnological processes, for the production of other valuable
metabo- lites. A solution for the treatment and disposal of
alpechin therefore should not only consider the environmental
aspects, but also the economic potential of valuable substances it
contains, including residual oil. In this case, income from
valuable products should make up for the cost asso- ciated with
treatment and disposal.
In Mediterranean countries, constraints imposed by the strict
envir- onmental protection regulations which will be enforced in
the EEC member states will also have be considered for proper
utilization of these effluents.
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Bioremediation of alpechin 253
The new perspectives for use of oil mill waters and olive
residues have been reviewed by different authors of various
countries such as Italy (Fedeli, 1986) Greece (Israelidis, 1990),
Tunisia (Friaa et al., 1986) and Spain (Ramos-Cormenzana,
1987).
In this article, some of the biotechnical solutions (processes)
proposed today for partial treatment and disposition will be
discussed briefly. Some of these processes are still at an
experimental level and have never been used on a large scale.
Although a fairly large number of potential uses of alpechin
have been proposed, in order to avoid going into details for all
the different aspects, we will only take into account the following
uses (Fig. 5):
(1) Its use as a fertiliser. (2) Its use as food and in the food
industry. (3) Its utilisation as a growth medium for algae. (4)
Biopolymeric substance production from alpechin focusing on
polysaccharide and biodegradable plastics. (5) Its use for
bioenergy production. (6) The employment of alpechin as a source of
biopharmaceuticals.
In one sense it can be said that the problem concerning alpechin
decontamination has been solved, but serious economical problems
still remain. Fedeli (1986) made an excellent study dealing with
the economic aspect of alpechin decontamination and utilization. He
considered five parameters: processing costs, investments, waste
cleaning costs, residual ecological costs, and products value. All
these factors need to be consid- ered to evaluate the feasibility
of the various processes that are proposed and to classify them
following an order of priority.
r\ :- . BIOPHARMAGEUTICALS
FERTILISER
bE :ENE)
FOOD AND THE BIOENERGETICS FOOD INDUSTRY
Fig. 5. Bioremediation of alpechin.
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254 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
The equation represents these factors is the following:
CE+CI+CD-RP=Q,
where:
CE = ecological costs; CI = industrial costs; CD = waste
cleaning costs; RP = product value; Q = profitability evaluation
factor.
Any kind of solution should take into account the above
considerations.
BIOREMEDIATION FOR USE AS A FERTILISER
The aqueous wastes from olive mill pressings represent a
valuable fertilis- ing material and a good source of organic
compounds, which hitherto have not been fully exploited, because
the wastes are produced in very large amounts during the brief
rainy season, they undergo rapid biode- gradation and are toxic to
plant and microbial life when applied to agri- cultural lands in
large quantities. The most prominent adverse effect is its
phytotoxicity that is largely attributable to the phenolic
compounds of alpechin. Indeed, when applied to soil, alpechin acts
like a broad-spectrum herbicide, where its half-life depends on the
amounts applied to soils and the frequency of application in a
given period on the soil aeration. In Spain, as we have previously
pointed out, it is illegal to dispose of the wastes directly into
waterways, and there are serious concerns that use of unremediated
alpechin as a fertiliser will cause pollution of water courses.
Use as a soil conditioner
As alpechin contains large amounts of minerals (potassium) and
organic matter (Fig. 6), its use as a fertiliser was tested after
prior neutralisation with lime and it gave satisfactory results in
experimental corn and wheat farming. The experiments carried out
indicated that it is advisable to use approximately 11 of
alpechin/m* (Albi Romero & Fiestas Ros de Ursinos, 1960).
Direct application of olive oil liquid waste onto soil was found
to stimulate the respiratory activity of the soil along with the
nitrogen-fixing capacity. Chatjipavlidis et al., (1986) reported
that repeat applications result in an enhancement of the microbiota
which can degrade the phyto- toxic components of the liquid wastes.
These authors claimed that their treatment could stabilize the soil
aggregates.
The effect of alpechin on the bacterial population of soil has
been investigated (Paredes et al., 1986; Moreno et al., 1987), and
also the
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Bioremediation of alpechin 255
ORGANIC MATTER 15% INORGANICS (1.8%):
POTASSIUM 0.85% SODIUM 0.13% NITROGEN 0.2%
COMPOSTING DIRECT APPLICATION
Fig. 6. Alpechin use as a fertiliser.
microbial feature of soil after pollution with oil mill waste
waters (Paredes et al., 1987) - a decrease in the count of aerobic
sporulated bacteria was noted.
No economic evaluation of this approach has yet been made since
it is still under investigation and has not reached the pilot plant
scale.
Although this approach is ecologically interesting to solve the
alpechin problem, unfortunately it does not allow the recovery of
the valuable substances contained in the liquid wastes.
In Spain, as ecological agriculture is becoming very popular, in
this sense alpechin conveniently prepared could be used as a
substitute for chemicals. The Spanish government is preparing new
environmental regulations that will probably include the
possibility of using fertilisers obtained from alpechin.
Our group is presently working on a project dealing with the
bior- emediation of olive-mill wastes for use as a fertiliser and
whose objective aims at understanding of the basic microbiological
and biochemical processes required to improve
recycling/detoxication composting proces- ses (Russell, 1994).
Physical and chemical processes to purify or detoxify alpechin
have met with limited practical success and were not shown to be
cost-effective. Microbial conversion appears to represent the only
viable alternative. Indeed, microbial isolates not only able to
resist the high osmolarity, elevated temperature and toxic nature
of alpechin, but which are also able to break down the major toxic
alpechin components, namely phenolics and fatty acids, have been
isolated. Moreover, the variety of organic components of alpechin
favors the development of microbial consortia endowed with a broad
array of metabolic capabilities that are useful in the
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256 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
bioremediation process of alpechin. The inclusion of
nitrogen-fixing species in the consortia has the added advantage
that the final product has a better mineral balance and thus a
greater fertilisation potential.
These investigations have shown that the compost obtained by
cornposting olive-mill wastes with other organic wastes can be used
as fertiliser and soil conditioner. All the alpechins are useful
for compost preparation, although their N and P contents were very
low, as was the level of micronutrients. Plant wastes (carriers)
can be quite suitable for preparing organic fertilisers, because of
their high organic content. Also, sufficient amounts of N were
found in some of the carriers (sugar cane sludge, spent mushroom
compost, cotton waste and grape mart). Humic- like substances are
generated during the alpechin cornposting, as a result of the
increasing humilication of the organic matter. The humic and fulvic
acids (HA and FA) concentration varied according to the method
used. The highest HA content was found in compost obtained by
mixing 654% of cotton waste with 34.6% of fresh poultry manure
(weight), with fresh alpechin at a rate of 1930 l/ton being added
to the solid mixture during the 18 days of cornposting. The lowest
levels of HA were found in compost obtained by mixing sewage sludge
with fresh alpechin. The HA content generally decreased in the
mature composts as compared with that found in the thermophilic
phase compost. The HA/FA ratio remained nearly constant during
cornposting [Cegarra, 1994, in Russells (1994) report].
Several reports suggest that nitrogen-fixing organisms increased
after soil treatment with olive-mill wastes (Garcia-Barrionuevo et
al., 1992, 1993). Therefore, it is likely that alpechin could be
used as a substrate for nitrogen fixation (Perez &
Gallardo-Lara 1987; Balis & Chatjipavlidis, 1994), and further
study should be of great interest.
However, although use of alpechin as a fertiliser appears a
viable alter- native, other bioremediation methods should be used
because this process alone would not be sufficient to cope with the
enormous quantity of alpechin produced.
USE AS FOOD AND IN BIOMASS AND SINGLE CELL PROTEIN FOOD INDUSTRY
(Fig. 7)
It is known that most research is either done in the
industrialised coun- tries, or is influenced by their needs; this
is the case of the alpechin substrate that could be used to produce
yeast for use as feed or fodder. The production of microbial
biomass for use as single cell protein will be economically viable
if the costs are low for large-scale production. Using Torulopsis
utilis, Fiestas Ros de Ursinos (1958) has shown that the yeast
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Bioremediation of alpechin 257
EDIBLE MUSHROOMS
SainzJtmenez 8 Gomez-Alarc6n. 1966
Fig. 7. Food and food industry uses for alpechin and
derivatives.
Codounlsel a., 1963
could grow sufficiently well in chemostat on alpechin to use the
process for industrial Torulopsis yeast production.
Mention should also be made of the observation that many kinds
of edible mushrooms are growing in composts fed with alpechin as
substrate (personal communications, Balis & Tomati).
In fact, the effects of alpechin on fungi has been investigated
by Saiz- Jimenez & Gomez-Alarcon (1986). They determine the
effect of alpechin on 19 imperfect fungi. With the exception of
Chaetomium &turn and Inotus hispidus, which were completely
inhibited, the other species studied were able to grow on malt
extract, although with a small delay with respect to the control
plates. Other studies have also been made on using olive milling
waste-waters as a medium for growth of Pleurotus spp. (Sanjust et
al., 1991).
The isolation of thermophilic fungi (Thermoascus, Mucor,
Tuluromyces, Torula, and Humicola) from co-composting material of
exhausted olive husk and oil mill waters has been reported by Demou
& Balis (1994). Although it needs to be confirmed, the same
authors have more recently isolated Coprinus species from similar
material (Russell, 1994).
Recovery of feed additives
Natural pigments (anthocyanins) contained in the alpechin have
been reported by several authors. Codounis et al., (1983) developed
a process for the extraction and purification of anthocyanins from
alpechin. According to this process the eluent from olive
extraction plants is first passed through an ultrafiltration unit
which separates the proteins and high molecular weight
carbohydrates from simple sugars, anthocyanins,
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258 A. Ramos-Cormenzana. M. Monteoliva-Sanchez, M. J. Lopez
phenolic substances, vitamins, aminoacids and minerals. The
concentrate, which is retained on the filter, is rich in proteins
and carbohydrates. It can be used, after fermentation, as a liquid
feed supplement or it may be dried and fed to animals.
The extraction and purification of anthocyanins contained in the
permeate can be achieved by chromatographic separation on ion
exchange resins. These anthocyanins can be used as natural food
colouring agents,
The antioxidant activity of alpechin phenols could also be
exploited for various applications in the food industry.
THE UTILISATION AS GROWTH MEDIUM FOR ALGAE
In a study on the microbial characterisation of effluent waters
from olive oil mill (Ramos-Cormenzana, 1986), it was found that a
large spectrum of microorganisms could grow on agar-solidified
medium made with the effluent waters as culture medium. It is thus
suggested that because of the high temperatures prevailing in
southern Spain, lagoons containing efflu- ent waters from olive oil
could be used to grow algae. Lagoon tempera- tures fluctuate
depending on liquid depth and ambient temperature. However, by
controlling the waste water flow in these lagoons it is expec- ted
that the temperature would be controlled. These lagoons could be
inoculated with species such as Dunaliella or Spirulina.
Ramos-Cormen- zana (1989) has already shown that Dunaliella can
grow on alpechin medium. The salinity prevents the invasion and
development of other microorganisms. Thus, the possibility is that
under seminatural condi- tions, or in ponds, Dunaliella could grow
in lagoons to produce biomass and b-carotene.
It has been suggested that biomass yields for Scenedesmus cells
grown in the Mediterranean area should be of the order of 30-32
g/m2 day (Tamiya, 1975). Efforts should be made to study the
ability of these algae to grow on alpechin medium in seminatural
conditions.
BIOPOLYMERIC SUBSTANCE PRODUCTION FROM ALPECHIN, FOCUSING ON
POLYSACCHARIDES AND BIODEGRADABLE
PLASTICS PRODUCTION
Ragazzi & Veronesse (1967) were the first to describe a
brownish-black coloured polymer present in alpechin. Since then,
the polymeric substances have been character&d in fresh
vegetation water and sludge evaporation ponds (Saiz-Jimenez et al.,
1986; Sanchez-Lama, personal communication).
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Bioremediation of alpechin 259
PHB (poly-hydroxy-b-butyric acid) and exopolysaccharides have
been detected in certain microbial cultures. Studies are in
progress in our laboratory to further characterize these
biopolymers (Fig. 8). Two of these polysaccharides are really
interesting: pullulan and xanthan.
Pullulan is an extracellular polysaccharide produced by the
fungus Aureobasidium pullulans. It is a marketable product of high
value and it is used in both food and pharmaceutical industries.
Olive oil liquid wastes were shown to provide good quality organic
substrate for pull- ulan production fermentation. Preliminary
experiments which were done at the Institute of Agricultural
Products Technology in Athens, showed that pullulan was produced at
yield of 8 g/l, with a ratio of bioconversion of total sugars of
O-5 g of pullulan/g total sugars, when the liquid wastes had been
diluted to an extent of three parts liquid wastes for one part
water.
Investigations in our laboratory on pullulan production has
provided a promising result. A relatively high conversion rate of
alpechin to poly- saccharide xanthan and pullulan were obtained.
The optimal alpechin concentration for growth and pullulan
production by Aureobasidium pull- ulans was 70% (vol/vol),
providing pullulan production yield above 3 g/L (Quevedo-Sarmiento
et al., 1991). A detailed study on xanthan formation in cultures of
Xunthomonus campestris, S4LLIA and Xanthomonas campestris, 646A
showed the ability of these microorganisms to grow and produce
xanthan in a medium containing 3&50% vol/vol alpechin
(Lopez-Lopez, 1993).
The use of alpechin as a substrate to exopolysaccharides
production by
0 PULLULAN j
HALOPHILE
A I
u!J
PHB
Fig. 8. Biopolymers obtained from alpechin.
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260 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
moderately halophilic microorganisms has also been assayed in
our laboratory. The rationale for these experiments was based on
the facts that several halophilic bacterial strain were shown to
degrade a variety of phenolic compounds (Yanase et al., 1992) and
that several moderately halophilic bacteria are shown to be good
producers of exopolysaccharides. Promising results were obtained
working with VolcunieZlu eurihdina, a moderately halophilic
bacterium described by us (Quesada et al., 1990), that produces an
extracellular polysaccharide in very large amounts, whose chemical
and physical properties could be of interest in a number of
industrial applications. The exopolysaccharide production was
investi- gated first using MY medium (Moraine & Rogovin, 1966),
and afterwards using MY medium modified by the incorporation of
different amounts of salts (up to 10%) and alpechin (up to 40%).
The EPS was isolated and purified, the EPS synthesis increased with
increasing salt concentration up to 7.5% (wt/vol); above this
concentration the productivity dropped drastically. In the absence
of salt, alpechin was also found to increase EPS production.
PHB and PHA
In a recent study on the effect of olive oil mill waste-waters
on Azoto- batter, it was observed that it stimulates PHB production
by Azotobacter chroococcum cells (Garcia-Barrionuevo, 199 1). More
recently, Martinez- Toledo et al. (1994) studied the PHB formation
from alpechin using Azotobacter chroococcum H23 strain. Therefore,
alpechin could most probably serve as an inexpensive substrate for
PHB, thereby increasing the likelihood of producing bioplastics at
competitive prices.
USE AS A BIOENERGETIC SOURCE (OR FOR BIOGAS PRODUCTION)
Biogas
Fermentation to produce biogas has been common practice in urban
waste water treatment plants for many years now. Compared to other
existing technologies, anaerobic treatment counts among those that
can reduce the amount of pollutants at a reasonable cost.
The final olive-mill waste waters, when tried as fertiliser, had
kept its phytotoxicity practically unchanged and, as described by
Ioniotakis et al. (1990), such highly toxic waste water cannot be
purified with biological methods. As with other types of anaerobic
treatment, methane and carbon
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Bioremediation of alpechin 261
dioxide are two of the degradation by-products. Alpechin is a
good potential energy source for anaerobic fermentation because of
its high organic content. Methane is a potential of biogas (Rozzi
& DiPinto, 1986; Carrieri et al., 1986); however, the
production costs for biogases are often rather high.
Methanogenesis is restricted to a group of prokaryotic
microorganisms which thrive in strictly anaerobic habitats, which
are designated metha- nogenic bacteria. Several studies have
already been carried out to evaluate the possibility of using
alpechin for methane production (Curi et al., 1988; Martin et al.,
1991).
The advantage of biogases is their further potential uses as
energy sources. For this reason biogas production processes were
retained among the research priorities at the Instituto de la Grasa
y sus Derivados, Sevilla.
Results obtained in Greece (Dal&, 1989; Georgakakis, 1989)
have shown that biogas was produced from olive mill waste wasters,
under anaerobic fermentation, giving yields of 1.5 m3 per m3 of
available volume in the bior- eactors. The fermentation resulted in
a sharp drop of the initial BODs where most of the major groups of
organic compounds were affected.
However, although substantial investments were made to try to
opti- mise this process, so far the results have been less
encouraging than was expected. One of the major problems is most
probably related to the influence of the alpechins, which are toxic
or inhibitory to microorganisms both under aerobic and anaerobic
conditions. The major effect of phenolic acids from alpechin on
microbial consortia involved in the methanogen- esis of olive mill
waters have been investigated (Sorlini et al., 1986).
Pretreatments of alpechin to reduce the phenolic compounds
through aerobic biodegradation processes using Aspergillus terreus
(Martinez- Nieto et al., 1992, 1993), or Azotobacter chroococcum
(Borja et al., 1993), were shown to substantially improve methane
production, when compared to methane production during anaerobic
digestion of untreated olive mill waste waters.
Production of ethanol
Production of other bioenergetic compounds from alpechin (Fig.
9) has also been evaluated. Butanol, butanediol (Wachner, Mendez
& Giolietti, 1988) and ethanol (Bambalov et al., 1989a, b)
counts among the compounds tested.
Ethanol can be produced using olive oil waste effluents as a
substrate. The microbial strains involved in these processes have
been isolated from such effluents and they were tentatively
identified as Candida wickerhamii, C. molischiana and Saccharomyces
cereviae (Bambalov et al., 1989a).
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262 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
IOENERGETICS
Fig. 9. Bioenergetics products obtained from alpechin.
The product yields (g of ethanol per g of sugar) reached 87% of
the theoretical yield, but the final alcohol concentration was low,
about 1.3% at the very best (Bambalov et al., 19893).
The recovery costs for such a low concentration of ethanol from
the fermentation broth are likely to be rather high if traditional
extraction methods are used. However, the process may be less
expensive if other methods such as membrane separation could be
used for extractions.
THE EMPLOYMENT OF ALPECHiN AS SOURCE OF BIOPHARMACEUTICALS
(BIOLOGICAL AND PHARMACOLOGICAL ACTIVITIES) (Fig. 10)
It is known that a number of compounds found in alpechin are
inhibitory for microorganisms (Gonzalez et al., 1990). Although the
use of such compounds as antimicrobials in pharmacological
applications has not yet been exploited, it is certainly to be
considered.
On the other hand, various alpechin components could be
extracted for specific applications. Polyphenols represent such a
class of compounds that could have interesting pharmaceutical
applications. We are aware that many research projects intended to
exploit these compounds are going on. More complex separation
strategies to recover alpechin compo- nents separately have been
suggested. Unfortunately information pertain- ing to these projects
is kept secret by the industries supporting those projects.
However, I will describe some of these useful applications,
based on published works, personal observations or personal
communications.
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Bioremediation of alpechin 263
ANTIBIOTICS
BIOLOGICAL RESPONSE MODIFIERS
BIOPHARMACOLOGICAL ACTIVITY1
NEWS PHARMACOBIOLOGICAL MOLECULES
Fig. 10. Biopharmaceutical uses of alpechin
The antibiotic production is one of the more feasible
applications, based on the known antimicrobial activity of
alpechin. More properly tetra- cycline could be among alpechin
components.
The polyphenols could be obtained from the alpechin and they
could be used in similar applications than other polyphenols
compounds that have been tested for their pharmacological activity.
Particularly they are good antifungal compounds. It has also been
suggested that these compounds could be used for the control of a
number of fruit and vegetable diseases (Lattanzio et al.,
1994).
Recently, vegetal polyphenols were considered as potential
antiviral compounds (Buciumeanu et al., 1994). Based on its
chemical composition I dare to comment that in a similar way,
alpechins polyphenols might also be considered for this
application.
In the last XVIIth International Meeting of the polyphenols
group, Takashi et al. (1994) pointed out that ellagitannin
oligomers have antitumoral activ- ities. Based on the relationship
between the whole structures of these oligo- mers and some that
could be found in alpechin, we suggested that it would be
worthwhile to study the possible application of oligomers from
alpechin as biological response modifiers. Similarly investigations
on polysaccharides obtained by alpechin biotransformation would be
interesting to initiate. The organic acids should also be studied
as biological response modifiers.
Other possibilities should not be excluded and enzyme inhibition
assays are valuable screening tools for the detection of the
bioactive action of alpechin extraction compounds. Their biological
activities are some times very difficult to detect by in vivo
assays due to the mildness of their actions. The separation and
extraction of some compounds could also be used in the industry in
the way that tyrosol and other pharmaceutical products have also
been studied.
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264 A. Ramos-Cormenzana, M. Monteoliva-Sanchez, M. J. Lopez
THE FUTURE
An interesting, relatively new proposal was suggested by
Israelidis (1990) for the treatment of bioremediation of
alpechin.
The method (in combination with a novel membrane developed at
the Research Centre KFA Julich of Germany in collaboration with the
firm ATP Ltd, Iraklion of Greece) offers the following
possibilities: (1) saving of water; and (2) recovery and production
of valuable substan- ces contained in the alpechin such as: (a)
fertilisers between 65 and 80 kg/m3 of alpechin including organic
and inorganic substances; (b) residual oil between 4 and 30 kg/m3
of alpechin; (c) colour substances (anthocyanines) 2-3 kg/m3 of
alpechin; (d) pullulan (polysaccharide) and/or ethanol in
quantities of 8 kg/m3 and 13 kg/m3 of alpechin, respectively.
This process had been primarily developed for sea water
desalination, but it was then trial-tested to treat alpechin in
Heraklion, Crete, and it appears to be very promising.
The point of issue is to develop an ecological and economical
way to dispose of olive oil production waste-waters, with a
simultaneous recovery of the water and of the valuable substances
contained in the alpechin (residual oil, fertiliser, chemical
compounds, fermentation by-products, etc.). The recovery and
production of useful substances should be done using
physicochemical and biotechnological methods.
These aims can be achieved with methods which detoxify alpechin
and separate the water and the valuable substances from alpechin in
order to obtain total recycling (excluding the residual oil).
Our own proposal is also based on the simultaneous recovery of
the water and the valuable substances contained in alpechin. We
suggest its use as composted fertiliser to extract chemical
compounds that could be applied in the biopharmaceutical industry,
and finally use of its biotrans- formation by-products such as the
biopolymers.
Although not completely, we have reviewed here the most relevant
applied systems and some considerations for future prospects.
We would like to point out that, although solutions are already
avail- able to dispose of alpechin, research efforts are still
required to exploit the additional potential values available from
alpechin.
ACKNOWLEDGEMENTS
The authors wish to thank the organisers for the opportunity to
contribute to this Symposium. Research in the authors laboratory
was supported
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Bioremediation of alpechin 265
partly by the Spanish Ministry (project AMB 92 723 CO3-01) and
the CEE (project EVWA-CT920006).
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