COMPOSITIONS COMPRISING SHIKIMIC ACID OBTAINED FROM OIL PALM BASED
MATERIALS AND METHOD OF PRODUCING THEREOF
FIELD OF INVENTION
The invention generally relates to a method to obtain and produce
shikimic acid from plants and plant based materials, and more
particularly to a method for producing shikimic acid based on oil
palm and oil palm based materials.
BACKGROUND OF THE INVENTION
Reference to any prior art in this specification is not, and should
not be taken as, an acknowledgement or any form of suggestion that
this prior art forms part of the common general knowledge in
Malaysia or any other country.
Shikimic Acid (SA) is an imperative biochemical intermediate in
plants and microorganisms, and has emerged as a vital starting
material in the synthesis of an anti-influenza drug, widely known as
Tamiflu (oseltamivir ). It is discovered to be highly functionalized
and it plays a major role in the production of essential aromatic
amino acids which are not synthesized in the human body, such as
phenylalanine, tryptophan and tryrosine. Presently, shikimic acid
can be obtained based on microbe-catalyzed synthesis schemes or from
plants, whereby the primary plant-based source for shikimic acid is
the star anise (Illicium anisatum) , whereby the yield of shikimic
acid from star anise seed can be as high as 7%.
Oseltamivir, also known as the trade name Tamiflu, aids in the
treatment and prophylaxis of both Influenzavirus A and
Influenzavirus B infection, by way of blocking the influenza virus
from spreading between cells and body. The administration of
oseltamivir is preferably conducted within 48 hours from the first
appearance of flu symptoms, as during this period it is observed
that the virus has yet to replicate and infect many cells thus
inhibition is considerably more effective. Under normal
circumstances, the compound from Tamiflu is hydrolyzed by esterases
in the liver, forming active form of drug, which is the oseltamivir
carboxylate. Oseltamivir carboxylate then binds to and thereby
inhibits viral neuralminidase . Such inhibition of neuraminidase by
Tamiflu and other neuraminidase inhibitors prevents the release of
new virus from the infected host cells.
It is observed based on current conditions that the primary
challenge in oseltamivir production is the availability of a viable
source of shikimic acid. As shikimic acid cannot be synthesized
economically, the notion in shortage of shikimic acid has been
surfaced due to increasing demand and consequently researchers began
bioprospecting in many areas, in their endeavours to find
alternative sources apart from star anise seed as briefly mentioned
above. Accordingly, shikimic acid content has been discovered in a
number of plants, particularly in the seeds of the sweetgum fruit,
the Indian plant Calophylium apetalum and Araucaria excelsea .
Nevertheless it has been observed that these plants produce toxic
compounds that necessitate prohibitively expensive refinement
methods to ensure and thus obtain purified shikimc acid.
It is being reported that at least 50 million people have been
treated with oseltamivir and this amount is expected to increase
within the upcoming years. It is further estimated that the market
size for Tamiflu will be at 400 million doses per year, at which
130mg of shikimic acid is required to synthesize one dose of
Tamiflu. From here, it can be expected that at least 52 metric tons
of shikimic acid is required to meet the upcoming demands. Although
production can be increased by way of replacing isolation from plant
sources with recombinant microbial catalysts following the need but
shortage of shikimic acid will intermittently be a challenging
factor especially in the occurrence of a pandemic worldwide.
Following this there is a great need to provide a renewable and
inexpensive source or viable alternative sources of shikimic acid.
The commercial value of oil palm ( Elaies guineensis) is in the oil,
which offers various health advantages and benefits, owing to its
high content of phytonutrients demonstrate antioxidant properties.
Nevertheless, at present, the oil palm based extracts, in particular
its vegetation liquour and palm oil mill effluent (POME) , has been
discovered by way of various studies to have beneficial implications
on skin ageing, cancer and heart diseases.
Malaysia is currently one of the world's largest producers of palm
oil. Being the major foreign exchange earner industry and to fulfill
the rapid progression of the world's demand for palm oil based on
the world population growth, Malaysia has at least 265 palm oil
mills all over the country.
Typically, the extraction of palm oil which is the primary product
of palm fruit, involves a combination of various processes, wherein
the main steps include the reception of fresh fruit bunches from
plantations, sterilizing and threshing of the bunches, mashing the
fruit and pressing the crude palm oil prior to storage and
exportation. However, the extraction of palm oil leads to the
production of oil palm vegetation liquor which is discarded in the
waste stream as Palm Oil Mill Effluent (POME) . In addition Empty
Fruit Bunches (EFB) and other waste materials are discarded in the
milling process
Palm o l mill effluent (POME) for instance, if untreated, is a
pollutant due to the high chemical oxygen demand (COD) and
biological oxygen demand (BOD) . The major components of POME include
oil and grease and solids, while elements found in POME include
phosphorus, potassium, boron, iron, manganese, copper, magnesium and
zinc. So far POME has been used in the preparation of fertilizers or
as a substrate for bacterial production of biodegradable plastics.
It is being reported that more than 500 kg (around 0.5 m3) of liquid
wastes, primarily in the form of palm oil mill effluent (POME) , are
discharged during the processing of 1.0 metric tons of fresh fruit
bunches (Ma et al. 1996) in a typical oil palm mill. Accordingly, it
is expected that more than 50 m 3 of POME from a mill after
processing 100 metric tons of fresh fruit bunches.
Proceeding from the above and based on recent studies on
phytochemicals extracted from oil palm, the palm oil industry
creates an abundance of prospect in beneficial phytochemicals and
other compounds recovery due to the amount of oil palm based
materials including waste materials generated from the industry, if
treated effectively.
It is a primary object of the present invention to provide a method
for production of shikimic acid based on oil palm extracts.
It is yet another object of the present invention to provide a
method for production of shikimic acid based on oil palm by
products .
It is another object of the present invention to provide a
composition and a method for producing a composition comprising
shikimic acid obtained from oil palm vegetation liquor.
It is yet another object of the present invention to provide a
composition and a method for producing shikimic acid obtained from
palm oil mill effluent (POME) .
It is yet another object of the present invention to provide a
composition comprising shikimic acid obtained from extracts from oil
palm-based materials and method for producing said shikimic acid
based on oil palm based materials.
It is yet another object of the present invention to provide a
composition comprising oil palm phenolics (OPP) with shikimic acid
content and method for producing said shikimic acid based on oil
palm based materials.
It is yet a further object of the present invention to provide a
composition and method for producing shikimic acid obtained from oil
palm liquid waste based materials.
Further objects and advantages of the present invention may become
apparent upon referring to the preferred embodiments of the present
invention as shown in the accompanying drawings and as described in
the following description.
SUMMARY OF INVENTION
There is disclosed a method for production of shikimic acid
comprising the step of obtaining extracts from oil palm-based
materials .
It is disclosed that the extracts comprises oil palm phenolics (OPP)
obtained from oil palm-based materials.
It is further disclosed that the shikimic acid are purified from the
OPP extracts.
It is further disclosed that the OPP extracts are obtained from palm
oil vegetation liquor.
It is further disclosed that the OPP extracts are obtained from palm
oil mill effluent (POME) .
In another aspect of the present invention there is disclosed a
composition containing shikimic acid, wherein said shikimic acid is
obtained from extracts of oil palm.
DETAILED DESCRIPTION OF THE INVENTION
In line with the above summary, the disclosed description and
examples relates to a composition comprising shikimic acid and
method of producing the said shikimic acid based on oil palm based
materials. It shall be apparent however to one skilled in the art
that the exemplifications are provided to better elucidate the
embodiments of the present invention and therefore should not be
construed as limiting the scope of protection.
As briefly described earlier, during oil palm fruit processing, the
extraction of palm oil generates a waste stream of vegetation
liquor, which is thereby considered as an aqueous phase of the
milling process prior to being discarded as palm oil mill effluent
(POME) and solid wastes.
In one embodiment of the present invention, a method for producing
shikimic acid comprises the steps of obtaining extracts from oil
palm based materials is disclosed. Suitably, it should be noted that
oil palm based materials including vegetation liquors, any oil palm
by products, palm oil milling wastes materials, oil palm based waste
materials (liquid and solid) , materials from oil palm based
industries, any ancillary oil palm vegetative matters, any part of
the oil palm tree for instance but not limiting to the stem, pulp,
seed, trunk, fruitlet, fruitbunch, frond, any part of the oil palm
fruit and other oil palm based materials and sources not name
explicitly herein.
The method further includes the step of obtaining extracts from oil
palm based materials preferably without using solvents.
In one embodiment of the present invention, the method may include
the step of obtaining water soluble, and enriched with phenolic
compounds, extracts from oil palm based materials.
In accordance with the present invention, the step of obtaining
extracts from oil palm based materials may be performed based on
conventional separation principles. It should be noted that however
the method involved for extraction is solvent-free.
According to the present invention, the method may further includes
the step of obtaining water soluble, antioxidant rich extracts known
as oil palm phenolics (OPP) from oil palm based materials including
vegetation liquors, any oil palm by products, palm oil milling
wastes materials, materials from oil palm based industries, any
ancillary oil palm vegetative matters, any part of the oil palm tree
for instance but not limiting to the stem, pulp, seed, trunk,
fruitlet, fruitbunch, frond, any part of the oil palm fruit and
other oil palm based materials and sources not name explicitly
herein.
Based on another embodiment of the present invention, the extracts
may be obtained from materials used or generated from oil palm based
industries .
In another embodiment of the present invention, the extraction of
the oil palm phenolics (OPP) as briefly described above in
accordance with a preferred embodiment of the present invention may
involve the steps of providing three-phase decantor system, and
preferably a plurality of separation stages using different types of
membranes suitably adapted to separate residual oil, ionic
contaminants and components of high molecular weight, thereby
obtaining oil palm phenolics (OPP) .
A s an example, oil palm phenolics (OPP) in accordance with an
embodiment of the present invention can be obtained from palm oil
mill effluent (POME) . Generally palm oil mill effluent (POME) is a
thick brownish liquid that contains approximately 4% solids by dry
weight. POME can be a source of nutrient content (Zakaria et. al)
upon treated to reduce the organic load (Ma et al.). Conventionally,
the treated POME may be recycled to the field. In one embodiment of
the present invention, shikimic acid is purified from said oil palm
phenolics (OPP) .
According to the present invention, the method further includes the
step of extraction or purifying shikimic acid from the oil palm
based materials extracts.
In accordance with a preferred embodiment of the present invention,
preparation of the extracts, oil palm or parts of oil palm may be
treated with glyphosate, a compound typically used in eliminating
unwanted plants especially weeds, so as to increase levels or obtain
high yield of shikimic acid. It is observed that glyphosate inhibits
the exnzyme EPSP synthase, thus condenses shikimate-3-phosphate and
phosphore-enol-pyruvate to form 5-enolpyruvylshikimate-3-phosphate,
a precursor of chorismate. It is understood that the dose of
glyphosate is suitably controlled to as to allow maximum yield of
shikimic acid.
It is understood that the extracts from oil palm based materials or
OPP extracts can be scaled up to accommodate larger or more
substantial extraction batch sizes desired.
In order to determine the radical scavenging activity of the OPP
prepared for the purpose of the present invention, standard
procedures may be carried out. Such may include, but not limiting
to, conducting Flash Chromatography or High Performance Liquid
Chromatography (HPLC) on the OPP extracts. Results showing the
compounds are obtained and presented in chromatograms . A free
radical scavenging activity based on the plotted graph of obtained
fractions and absorbance may be provided.
A s an example, HPLC was carried out on a sample of OPP extracts.
Results obtained to show the presence and HPLC profile of shikimic
acid are as shown in FIG 1 .
In accordance with a preferred embodiment of the present invention,
the determination of elements within the extracts can be carried out
using various standard procedures, for instance, but not limiting
to, mass spectroscopy.
A Nuclear magnetic resonance (NMR) analysis may further be carried
out to confirm such purification.
In an embodiment of the present invention, extraction and
purification of shikimic acid from OPP may be conducted based on
standard purification methods. Suitably, it is understood that the
extract and purification can be scaled up to accommodate larger or
more substantial extraction batch sizes desired.
According to the present invention, the yield of shikimic acid
obtained from OPP is approximately 1% of the dry weight. It is
expected that the amount may increase if plant is treated with
glyphosate prior to extracting the OPP.
An example of estimates of the amount of palm oil mill effluent
(POME) may be required for production of shikimic acid at different
extraction efficiencies are as depicted in TABLE 1 below:
Apart from being the primary building block in the production of
Tamiflu, shikimic acid has been reported based on various recent
5 studies, to provide other various health related benefits.
Shikimic acid obtained based on the present invention may be used as
an immunomodulatory agent. As an example, in an in-vitro model,
incubation of peripheral blood mononuclear cells with shikimic acid
10 in combination with the polyphenol quercetin led to an increase in
the release of chemokines IL-8 and IL-6, whereby these chemokines
play a major role in recruiting and activating lymphocytes for the
innate immunity system. Such discovery therefore demonstrates that
shikimic acid and quercetin from dietary may enhance the innate
15 immunity system, which would be beneficial for combating viral
infections (Bertelli et.al) .
In accordance with the present invention, the shikimic acid obtained
may be used in the treatment for hair loss. As an example, an
experiment was conducted and water soluble extracts containing
shikimic acid obtained from Illicium anisatum (aniseed) has been
reported to be able to aid in increasing subcutaneous blood flow and
in mice and thus stimulate the elongation of mouse follicles in
culture (Sakaguchi et. al .)
Further in accordance to the present invention, the OPP extracts
containing shikimic acid obtained from oil palm based materials may
be used to make, or are contained in but not limiting to, drinks,
edible products, health supplements, antioxidant additives,
cosmeceutical products or medicinal products.
From the foregoing it is disclosed that oil palm based materials can
be considered as the most abundant and inexpensive source for
shikimic acid. It is estimated that 46 million tons of palm oil mill
effluent (POME) can be produced yearly.
It is understood by a person skilled in the art that the methods for
experiments and studies are described as exemplifications herein and
thus the results are not intended, however, to limit or restrict the
scope of the invention in any way and should not be construed as
providing conditions, parameters, agents or starting materials which
must be utilized exclusively in order to practice the present
invention. It is therefore understood that the invention may be
practiced, within the scope of the appended claims, with equivalent
methods for the experiments than as specifically described and
stated in claims.

CLAIMS
1 . A method for production of shikimic acid comprising the step
of obtaining extracts from oil palm-based materials.
2 . A method as claimed in Claim 1 further comprising purifying
said shikimic acid from said extracts from oil palm-based
materials .
3 . A method as claimed in Claim 1 wherein the extracts are
obtained from oil palm by- products.
4 . A method as claimed in Claim 1 wherein the extracts are
obtained from oil palm based waste materials.
5 . A method as claimed in Claim 1 wherein the extracts are
obtained from oil palm liquid waste materials.
6 . A method as claimed in Claim 1 wherein the extracts are
obtained from palm oil vegetation liquor.
7 . A method as claimed in Claim 1 wherein the extracts are
obtained from materials from oil palm based industries.
8. A method as claimed in Claim 1 to 4 wherein the extracts are
obtained from palm oil mill effluent (POME) .
9 . A method as claimed in Claim 1 wherein the extracts are
obtained from anyone of the following oil palm based
materials; oil palm by products, palm oil milling wastes
materials, materials from oil palm based industries, any
ancillary oil palm vegetative matters, any part of the oil
palm tree for instance but not limiting to the stem, pulp,
seed, trunk, fruitlet, fruitbunch, frond, any part of the oil
palm fruit and other oil palm based materials and sources.
A method as claimed in Claim 1 wherein the extracts
comprise of oil palm phenolics (OPP) .
11. A method as claimed in Claim 1 wherein said extracts are
water soluble and enriched with anti-oxidants .
12. A method as claimed in Claim 1 wherein the method further
comprising treating said oil palm with glyphosate prior to
obtaining the extracts from said oil palm-based materials.
13. A composition containing shikimic acid, wherein said
shikimic acid is obtained from extracts of oil palm or oil
palm based materials.
14. A composition as claimed in Claim 13 wherein the extracts
comprise of oil palm phenolics (OPP) .
15. A composition as claimed in Claim 13 to 15 wherein said
shikimic acid is obtained from palm oil mill effluent (POME).
16. A composition as claimed in Claim 13 wherein the shikimic
acid is obtained from palm oil vegetation liquor.