BIODIVERSITAS ISSN: 1412-033X (printed edition)

Volume 10, Number 3, July 2009 ISSN: 2085-4722 (electronic)

Pages: 151-157 DOI: 10.13057/biodiv/d100309

Extraction of Coconut Oil (Cocos nucifera L.) through

Fermentation System
♥
RINI HANDAYANI , JOKO SULISTYO, RITA DWI RAHAYU
Microbiology Division, Research Center for Biology, Indonesian Institute of Sciences (LIPI), Cibinong-Bogor 16911

Received: 28th October 2008. Accepted: 4 December 2008.

ABSTRACT

Coconut oil (Cocos nucifera L.) has a unique role in the diet as an important physiologically functional food. The health and
nutritional benefits that can be derived from consuming coconut oil have been recognized in many parts of the world for
centuries. There are few techniques for coconut oil extraction, such as physical, chemical, and fermentation or enzymatic
processes using microbial inoculum as enzymatic starter. Starter with different concentration (1.0; 2.5; 5.0; and 10%) of
microbial strains were added into coconut cream and allowed to be fermented for over night. The extracted oil was analyzed
for further experiment, especially on its antibacterial activity. The maximum yield of 27.2% was achieved by adding 5.0%
starter. Water content, acid value, FFA, and peroxide value of the fermented coconut oil were 0.3%, 0.45%, 0.22% and
2.54% respectively. A gas chromatogram showed that this fermented oil contained high lauric acid (46.82%), and 6.01%
caprylic, 7.5% capric, 17.02% miristic, 7.21% palmitic, 3.11% palmitoleic, 5.41% stearic, and 1.3% linoleic acid,
respectively. An inhibitory effect of such kind coconut oil which contains potential fatty acid against bacterial growth was
further examined. It was found that this edible oil exhibited antibacterial activity to inhibit the growth of Bacillus subtilis,
Escherichia coli, Pseudomonas fluorescence, Bacillus cereus and Salmonella; however it showed slightly inhibitory effect
when it was exposed to Bacillus cereus and Escherichia coli.
© 2009 Biodiversitas, Journal of Biological Diversity

Key words: coconut oil, inoculum, fermentation, lauric acid, antibacterial.

INTRODUCTION part of their diet have a low incidence of health

problems associated with blood clotting, including
A novel method on improving quality of coconut oil heart disease and stroke (Prior et al., 1981). Coconut
(Cocos nucifera L.) to produce best quality of coconut oil is very stable and does not need to be refrigerated
oil for industrial application is increased. One of them since it contains a saturated fatty acid, because all
is a method of extraction through fermentation or the carbon-atom linkages are filled or saturated with
enzymatic system (Rosenthal et al. 1996). Virgin hydrogen. This means that they do not normally go
coconut oil (VCO) is made from fresh coconuts, not rancid, even when they heated, degraded, irradiated,
copra. Since high temperatures and chemicals oxygenated for cooking or other purposes (Issacs,
solvent are not used, the oil retains its naturally 1986; Rindengan and Novarianto, 2004).
occurring phyto-chemicals which produce a distinctive The VCO processed by fermentation or enzymatic
coconut taste and smell. The oil is pure white when system has more beneficial and safety effect rather
the oil is solidified, or crystal clear like water when than traditional methods from copra, since they often
liquefied. The oil contained high lauric acid (C-12, c.a. infected by insects or aflatoxin producing molds that
50%) as saturated fatty acid and has known well as caused potential toxicity problem during
medium chain fatty acid (MCFA). MCFAs are burned manufacturing. Traditional coconut oils are
up immediately after consumption and therefore the considered to be low quality products which indicated
body uses it immediately to make energy rather than by high moisture and free fatty acid content. It was
store it as body fat (Enig, 1996; Kabara, 1984). therefore easily to rancid and turned to brown and
Studies have revealed that populations who exhibited relatively short life-time by sensory test
traditionally consume large quantities of coconut as a (Soeka et al., 2008).
Extraction process of coconut oil through
fermentation or enzymatic system involved microbial
starter inoculums or enzymatic starter that play a role
 Corresponding address: on breaking of coconut milk emulsion, while through
Jl. Raya Bogor Km. 46 Cibinong-Bogor 16911
Tel. +62-21-8765066. Fax. +62-21-8765063/62 traditional processes the oil extraction were carried
e-mail: handayanirini@yahoo.co.uk out physically by using of heating or mechanical
152 B I O D I V E R S I T A S Vol. 10, No. 3, July 2009, pp. 151-157

expelling (Ketaren, 1986). Activities of enzymes were Agar media preparation

affected by substrate and enzyme concentration, pH, Potatoes dextrose agar (PDA) media and nutrient
temperature, and incubation time (Pelczar and Chan, agar (NA) media containing 0.75 g yeast extract, 1.25
1986). Microbial starter was utilized as their shown on g peptone, 5g agar and 10g potato or malt extracts,
proteolytic, amylolytic and lipolytic capacities. These respectively, were prepared according to Cappuccino
kinds of enzymes are required to hydrolyze protein, and Sherman (1983). These ingredients were
carbohydrate and lipid components contained in the dissolved into 250 mL distilled water, and then melted
coconut kernel. Basically, the purpose of the using microwave for 3 min to accelerate their
fermentation or enzymatic processes is to make the solubility. The melted media were poured into tubes
coconut emulsion into unstable condition and and autoclaved for 15 min at 121ºC and cooled down
therefore easily to separate into oil phase on upper onto elevate rack to prepare slant culture media.
layer and carbohydrate, protein and water phase on
below layer (Soeka et al., 2008; Rahayu et al., 2008). Microbial screening
The purpose of research was to develop the To prepare enzymatic starter for extracting virgin
extraction method of coconut oil to produce high coconut oil, both of yeast and mold strains were
quality virgin coconut oil (VCO) and expectedly useful inoculated onto PDA and bacterial strain onto NA and
for improving technology on extraction of VCO then incubated for 3 days at room temperature. Stock
naturally. cultures were transferred into liquid media containing
coconut water, coconut skim, pineapple or malt
extract, urea and molasses.
MATERIALS AND METHODS
Assay for enzymatic activities
Microbial strains
Selected media for assaying proteolytic and
The strains used in this experiment were
amylolytic activities was referred to a method of
Lactobacillus bulgaricus, Saccharomyces cerevisiae,
Sulistyo et al. (1999). One ose-needle of stock culture
Candida rugosa, Aspergillus oryzae, Salmonella,
of bacterial strain was inoculated into nutrient broth
Pseudomonas fluorescens, Escherichia coli, and o
(NB) and incubated for 24h at 37 C. One mL of
Bacillus substilis, soy-sauce starter (Aspergillus
culture was added into 9.0 mL of NB media and
oryzae), bake yeast (Saccharomyces sp), tempeh o
incubated for 24h at 37 C. One mL of respective
starter (Rhizopus oligosporus), and beverage yeast
stock cultures were inoculated into 9.0 mL NB, and
(Candida utilis) obtained from the collection of o
incubated at 37 C for 2 days. Proteolytic activity was
Microbiology Division of Research Center for Biology,
measured semi quantitatively on the media containing
Indonesian Institute of Science (LIPI) Cibinong-Bogor.
1% KH2PO4, 2% MgSO4.7H2O, 1% yeast extract, 5%
agar and 2% skim milk. Qualitatively analysis was
Chemical reagents
done based on activity tested on agar media grown
Bacto-peptone, yeast extract, agar, potatoes
with 3 days-old microbial strains. The proteolytic
dextrose agar (PDA), KH2PO4, MgSO4.7H2O, soluble
activity was indicated by present of clear zone
starch were purchased from Sigma and Merck.
surrounded colonies of strains. Amylolytic activity was
measured as by measuring proteolytic activity when
Steps of experiment 1% soluble starch was applied to the media rather
The experiment was carried out on five steps, i.e.: than 2% skim milk. Observation was carried out on
(i) strains selection, (ii) starter production, (iii) coconut present of clear zone after employing iodine reagent
oil extraction, (iv) qualitative and quantitative analysis (Mestecky et al., 2004).
of oil product, (v) assay on antibacterial activity of
coconut oil. Enzymatic starter preparation
The media for production of starter containing
Coconut oil preparation coconut water, coconut skim, pineapple or malt
The coconut type used for making virgin coconut extract, urea and molasses in 500mL Erlenmeyer
oil was according to method of Rindengan and flask was sterilized using autoclave for 15 min at
Novarianto (2004). The coconut cultivars used in this 121ºC. After cooling down to a room temperature, the
experiment were kelapa dalam, genjah salak, and media were inoculated by Lactobacillus bulgaricus,
genjah kuning. Matured coconut was grated and the Aspergillus oryzae, Candida rugosa and
grated coconut was then mixed with hot tap water Saccharomyces cerevisiae, respectively. The
(1:1, w/v). After squeezing and filtering, coconut milk respective starters those were incubated with different
was pooled into a clean jar and stayed for 1h. After cultures were then employed to the coconut cream
separating into two layers those were cream on upper o
and incubated at 40 C for overnight. The oil was
part which riched in oil content while skim layer riched obtained through this process were then measured
in protein on below part was drained off, and the and analyzed. Influence of strains growth toward
remained cream one was then fermented overnight to incubation temperature at 25, 30, 35, 40 and 45 C
o
prepare virgin coconut oil. and pH of media at 3, 4 , 5, and 6 during incubation
 HANDAYANI et al. – Extraction of virgin coconit oil 153

on the shaker for 5 days and employed concentration A = Quantity of NaOH

of starter at 1%, 3%, and 5% toward the yield of N = Normality of NaOH
obtaining coconut oil were studied to determine M = MW of lauric acid
capacity of respective microbial strains those were
suitable for preparing an effective starter in producing Moisture content
high yield of VCO (Sulistyo et al., 1999). Moisture content was determined by weighing 10g
The strain that exhibited high yield on oil production of sample and placed onto a petri dish that had
was selected and examined further in comparison to already determined for its blank weight. The sample
the capacities of commercial starter products on was heated at 105ºC for 2h in an oven and cooled it
extracting coconut oil through fermentation system, down in a desiccator for approximately 15 min and
such as soy-sauce starter mold (A. oryzae), bake weighed again.
yeast (Saccharomyces sp.), tempeh starter (R.
oligosporus), and alcoholic beverage yeast (C. utilis). Moisture content = A - B x 100%
The obtaining oils derived from different coconut type A
were then filtered through activated charcoal and
analyzed using gas chromatography (GC). Assay on antibacterial activity
Antibacterial activity was assayed by preparing
Yield measurement nutrient broth (NB) in some reaction tubes. The media
Yield of obtaining coconut oil was determined by containing 0.3 g yeast extract and 0.5 g peptone in
o
using the method of gravimetric (v/v) as follows: distilled water was sterilized by autoclaving at 121 C
for 15 min. The NB media was incubated on the
Yield = Volume of obtaining oil (mL) x 100% shaker after inoculating with 1ose of tested bacteria
Volume of coconut cream (mL) for 2 days. One mL of pre-incubated media which
containing the tested bacteria was then diluted into
Fatty acid analysis the tubes containing 9.0 mL of sterilized distilled
Sample of VCO was analyzed according to the water and more over diluted gradient up to obtaining
-3 -3
method of Rietschel et al. (1972). Approximately 20- dilution at 10 . Finally, 0.1 mL of the 10 diluting
30 mg of sample was placed into a tube with cap and sample was transferred onto the petri dish containing
added with 1.0 mL 0.5 N NaOH in methanol and NA media. To determine the activity of VCO against
hydrolyzed for 20 min. After addition with 2.0 mL of bacterial growth, a smeared paper dish with VCO was
16% BF3 in methanol and 2.0 mL of saturated NaCl to placed onto the media, and the activity of antibacterial
remove emulsion, the reaction mixture was then was assayed by observing the present of clear zone
extracted with hexane. The hexane layer was then surrounding the colony that had grown by tested
transferred into a flask containing anhydrate 0.1g strains after 2 days incubation (Carson and Riley,
Na2SO4 as moisture absorbent. The prepared sample 1995).
was injected onto GC using internal standard of fatty
acids, under GC condition at 190-200ºC, flow rate 1.0
RESULTS AND DISCUSSION
cm/s, fused silica capillary column (3 m length), flame
ionization detector and volume of sample injection
was 4.0 µL. Amylolytic and proteolytic activity
To determine the capacity of some microbial
Proximate analysis strains in producing enzymatic starter that was
suitable for extraction of VCO, we had employed four
According to the method of Suminar et al. (2001),
selected microbial strains were L. bulgaricus, S.
the proximate analysis for determining acid value,
cerevisiae, C. rugosa and A. oryzae. The enzymatic
free fatty acid, moisture and peroxide content of VCO
was carried out by preparing 2.5 g of sample onto activities of these strains were investigated according
erlenmeyer flask. A titration reagent of 25 mL alcohol- to the method of gel diffusion on the media containing
starch for amylolytic activity or skim milk for
benzene (1:1, v/v) was pre-heated on a water bath at
proteolytic activity as mentioned in the Methods and
70ºC for 10 min after addition with 3 drops of
Materials. The strain of L. bulgaricus showed the
phenolphtalein as indicator and the mixture was
highest activity for amylolytic and proteolytic enzymes
titrated with 0.01 N NaOH up to the solution just
turned to slight red. The solution was mixed with as indicated by formation of colony surrounding clear
sample and heated for 5 min and titrated again with zones. Diameter of clear zone ( 2.0 cm) was
0.01 N NaOH at least for 10 min. undoubtedly illustrated that the strain of L. bulgaricus
capable to produce amylase and protease those were
Acid value = A x N x 40 availably important to digest protein and carbohydrate
Sample weight (g) which contained in coconut cream as its substrate.
The strain L. bulgaricus was furthermore selected to
FFA (%) = A x N x M x 100% be employed as potential starter for extracting VCO,
Sample weight (mg) while the other strains, S. cerevisiae, C. rugosa and
A. oryzae, respectively. A. oryzae had not been
154 B I O D I V E R S I T A S Vol. 10, No. 3, July 2009, pp. 151-157

employed as starter since their proteolytic and coagulate protein in consequence of phases
amylolytic activities given are lower than L. bulgaricus formation of oil on upper part, protein in the middle
(Table 1). and water layer on lower part (Rindengan and
Novarianto, 2004). Due to a lower molecular weight,
Table 1. Proteolytic and amylolytic activities of the selected the oil part formed through the process could be
strains. directly separated from protein and water part by
draining off both of them through a valve (Figure 1.B).
Diameter of clear zone (cm) To reduce interference of water content or insoluble
Activity/Strain
LB K-1A SC CR
materials into the oil part, a further process of
Proteolytic 2.0 1.6 1.3 1.6
obtaining oil by refining through filter paper or vacuum
Amylolytic 2.1 1.4 1.4 1.5
Note: LB: Lactobacillus bulgaricus; K-1A: Aspergillus oryzae; filter and rinse with hot water following by vacuum
SC: Saccharomyces cerevisiae; CR: Candida rugosa. evaporation was required to avoid chemically
processing to achieve the virgin state of oil as shown
Fermentation system on Figure 1.C.
The fermentation of coconut cream occurred when Fermented coconut oil has been known well as
the enzymatic starter had been employed for virgin coconut oil (VCO) since high temperatures,
processing. Crude coconut oil was formed due to a chemicals or other physical treatment are not used in
phenomenon of protein digestion that plays a role to its processing. As it had been naturally and
stabilize emulsion of the coconut cream into a soluble traditionally processed through enzymatic
material. The enzymatic starter with high capacity of fermentation, unhydrogenated, undeodorized, and
amylolytic and proteolytic could hydrolyze unbleached, the component of fatty acids, especially
carbohydrate and protein which contained in the lauric acid of this coconut oil is not change since it is
coconut the cream as its substrate into soluble sugar least vulnerable of all the dietary oils to oxidation and
and amino acid and peptide (Soeka et al., 2008). The free-radical formation, and it is therefore the safest to
extraction process of coconut oil via fermentation or use in cooking. It does not become polymerized and
enzymatic system involved microbial cell and form by-products as do other oils when heated to
enzymes those could solve the emulsion; however, normal cooking temperatures (Kaunitz and Dayrit,
their activities were influenced by some conditions of 1992; Rindengan and Novarianto, 2004; Sulistyo,
substrate, enzyme, pH, temperature, and incubation 2004).
period (Pelczar and Chan 1986). Structurally, coconut oil is very rare amongst all
Preliminary step on extraction process of VCO the other dietary lipids. As a different class of
was initiated after separating the coconut cream saturated fat that behaves very differently in the body
which higher in lipid content from coconut skims from each other, since it is composed almost entirely
which higher in carbohydrate and protein content as of medium chain fatty acids (MCFA), a powerful anti-
shown on Figure 1.A. After addition with starter microbial, where mother's milk is very high in
followed by overnight fermentation of the coconut them. Coconut oil is composed of an incredible 64%
cream at room temperature, the starter containing MCFA. The body metabolizes MCFA and absorbed
enzymes were stimulated to digest starch and directly from the intestine into the portal vein, and
ferment it into alcohol and organic acids that sent straight on to the liver, where they are burned for

A B C

Figure 1. A. Coconut milk separation, B. Fermentation process, C. Purified coconut oil.

 HANDAYANI et al. – Extraction of virgin coconit oil 155

fuel, almost like a carbohydrate. Rather than produce the concentration of substrate achieves optimal
fat, they are used to produce immediate energy. And, condition since the enzyme is saturated by the
the body uses much less energy to digest MCFA. substrate binding enzyme complexes.
They are easily digested by saliva and stomach
enzymes and do not require pancreatic enzymes.
This relieves stress on both the pancreas and 30

Yield of Oil (%)

the digestive system. For this reason, MCFA are 25
20
essential in baby formulas, and are routinely used in 15
hospitals for patients with digestive, metabolic and 10
malabsorption problems. Also, the MCFA in VCO are 5
0
used to improve insulin secretion and the utilization of
LB 3 K-1A 4 CR 5 SC 6
glucose, and therefore greatly helps relieve the pH of Starter
symptoms and reduce the health risks of diabetes
(Enig, 1996).

Screening on microbial strains Figure 2. Effect of pH of starter on yield of extracting oil.

Influence of employing pH (Figure 2), temperature
(Figure 3) and starter concentration (Fig 4) on the
yield of extracting coconut oil exhibited that the strain

Yield of Oil (%)

of L. bulgaricus could effectively extract the oil higher 30
than the tested microbial strains when the starter was 20
employed to incubate the coconut cream under the
º 10
fermentation condition at pH 5.0, 45 C and 5% starter
concentration. 0
Figure 2 showed the influence of incubation pH on 25 30 35 40 45
LB K-1A
CR SC
the yield of extracting oil. It was found that the highest Incubation Temperature ( C) 0

yield of oil (27.0%) could be obtained after incubating

the starter at pH 5.0. It is probably that the strain of L.
bulgaricus which employed as the starter is a Figure 3. Effect of temperature on yield of extracting oil.
facultative aerobic strain that had optimal proteolytic
capability at pH 5.0. This pH value is affected on
occurring toward substrate binding enzyme since the
+
concentration of H potentially affected a linkage 30
Yield of Oil (%)

between active site of enzyme and its substrate which

20
led to conform the active site into optimal condition for
binding the substrate based on the principal of Lock 10

and Key. 0
Figure 3 showed the influence of incubation LB 1K-1A CR 3 SC 5
temperature on the yield of extracting oil. It was found Starter Concentration (%)
that the highest yield of oil (27.2%) could be obtained
o
after incubating the starter at 45 C. It is probably that
the strain of L. bulgaricus behaves at optimal
o Figure 4. Effect of starter concentration on yield of
condition between 40-45 C. The binding of enzyme to extracting oil.
its substrate and rising temperature up to a certain
degree had increased kinetic energy and promoted
movements of reacted molecules. It was therefore Moreover the starter capability of L. bulgaricus in
increasing bumping occurrence between enzyme and extracting of VCO was investigated furthermore in the
its substrate optimally. The enzyme exhibited its comparison to other strains contained in soy-sauce
activity at certain optimal condition of temperature, starter mold (A. oryzae), bake yeast (Saccharomyces
and therefore when the temperature is over than its sp), tempeh starter (R. oligosporus), and alcoholic
optimal condition, the enzyme would certainly be beverage yeast (C. utilis). To produce kinds of VCO
denaturated. products, the coconut milk was prepared by different
Figure 4 showed the influence of starter strain of coconut cultivars such as kelapa dalam of
concentration on the yield of extracting oil. It was Cianjur Regency, genjah salak and genjah kuning of
found that the highest yield of oil (26.8%) could be Bogor Regency. The yield of oil derived from each
obtained after incubating the starter at 5.0% (v/v). At types of coconut were then purified by filtration and
low concentration rate of reaction was too low, absorption using absorbent of activated charcoal as
however, the rate would be higher as increasing of shown on Table 2.
substrate concentration which catalyzed by the The Table showed that the highest yield of oil was
enzyme. Increasing of enzyme is not effective when obtained by using coconut cream of kelapa dalam
156 B I O D I V E R S I T A S Vol. 10, No. 3, July 2009, pp. 151-157

Table 2. Yield (mL) of VCO derived from different type of (Soeka et al., 2008).
coconut.
Quantitative analysis of VCO
Coconut VCO Kecap Tempeh Beverage Bake
To determine the content of FFA, moisture
Cultivars starter starter starter yeast yeast
content, peroxide value, the extracting VCO obtained
Kelapa dalam 25.5 28.0 21.0 - -
Genjah salak 21.5 23.0 18.0 - - by using of the starter of L. bulgaricus was then
Genjah 15.5 17.0 15.0 - - analyzed and showed as Table 4.
kuning
Note: (-) none oil formed after processing. Table 4. Analysis of VCO according to SII. 0150-72 for
edible oil.

(24.83 mL) while using genjah salak and genjah Component of Reference Concentration
kuning gave lower yield of oil was 20.83 mL and Analysis Value of SII (%)
15.83 mL, respectively. It was apparent that initial Moisture content Max 0.5 % 0.30
composition of coconut cream significantly influence Acid value Max 0.5 % 0.45
on final yield of extraction oil. The coconut cream of Free fatty acid Max 2.5 % 0.22
kelapa dalam exhibited calories (359 cal) and lipid by Peroxide value Max 3.0 % 2.54
means of oil content (34.7%) higher than both of the
genjah types of coconut cultivar those were 180 cal
and 13.0% on the average (Palungkun, 1993). It was Table 4 showed that characteristic of obtaining
found that the oil layer was formed on the coconut VCO for edible oil referred to the moisture content,
cream was incubated by the starter of L. bulgaricus, acid value FFA and peroxide values was agreed with
A. oryzae of soy-sauce and R. oligosporus of tempeh a range of value in accordance to the Standard
inoculum, while there was none of oil layer found on International of Indonesia (SII) 0150-72 for edible oil.
the coconut cream was incubated by Saccharomyces It was suggested that our VCO which was extracted
sp. of bake and C. utilis of beverage starters. through enzymatic fermentation process by
Apparently, the enzyme of these kinds of starters had employing the starter of L. bulgaricus was
been inactivated or not appropriated as the starter for appropriated to be consumed as safety and healthy
enzymatic fermentation process of coconut oil, and edible oil. The quality of the obtaining oil was
resulted in none oil formation since the coconut corresponded to requirement for quality standard of
cream as the substrate was not agreed with the good edible oil. One of spoiled edible oil indicator is
enzymes contained in such starters. high in acid and peroxide values, since their
existences in the product indicate an alteration
Fatty acid analysis of VCO caused by oxidation on chemical content is being
To determine fatty acid composition of the virgin occurred and resulted frequently in a problem of
coconut oil (VCO) obtained by the enzymatic rancid.
fermentation, sample of the oil was then analyzed by
using GC. It was found that the highest yield of lauric Assay of antibacterial activity
acid of oil (42.95%) was obtained by employing the The antimicrobial properties lauric acid and its
starter of L. bulgaricus into coconut cream derived derivative monolaurin from coconut oil have shown
from kelapa dalam strain as shown on Table 3. promise in this study. Lauric acid, which is present in
high concentration in coconut oil, forms monolaurin in
Table 3. Analysis of lauric acid of extracting VCO. the animal body and this derivative of lauric acid can
inhibit the growth of pathogenic microorganisms
Lauric acid concentration (%) (Kabara, 1984). The research focused on
Fatty Acid
VCO-LIPI AO–D LB-D AO-GS LB-GS Pseudomonas fluorescence, Bacillus substilis,
Lauric acid 46.82 41.46 42.95 40.68 35.08 Salmonella and Escherichia coli. To determine
potential of lauric acid contained in this extracting oil
obtained through enzymatic fermentation, this oil was
Table 3 showed that the VCO obtained by then studied furthermore against microbial growth. Its
employing the starter of L. bulgaricus into coconut antimicrobial activity was observed by the existence
cream of kelapa dalam (LB-D) yielded higher lauric of clear zone formed surrounding paper-disc that had
acid content (42.95%) rather than by employing the pre-submerged into this oil on the media grown with
starter of A. oryzae with strain of kelapa dalam (AO- colonies of tested microbial strains. It was found that
D, 41.46%) and the starter A. oryzae with the strain of the clear zone surrounding the paper disc on the
genjah salak (AO-GS, 40.68%) or the starter of L. media fully grown with strain of Salmonella, indicated
bulgaricus with the strain of genjah salak (LB-GS, that this oil had activity against the growth of tested
35.08%). It was found somehow; the yield of strain as shown on Figure 5. It is now clear and
obtaining oil still lower yet rather than the oil had been scientifically validated that the inclusion of coconut oil
obtained by using the starter of VCO-LIPI as the in the diet could and should be utilized for its
standard of comparison of previous experiment preventive and healing properties.
 HANDAYANI et al. – Extraction of virgin coconit oil 157

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Standard International of Indonesia (SII) 0150-72. To Penerapan teknologi fermentasi pada bioproses fermentasi
determine potential of lauric acid contained in this oil, minyak kelapa (fermikel). Berita Biologi 4 (5): 273-279.
the study was focused on some microbial strains and Sulistyo, J. 2004. Pemasyarakatan, Penerapan, dan
It was found that there were clear zone surrounding Pengembangan TTG. Lokakarya TTG-Badan Pemberdayaan
Masyarakat dan Penanggulangan Sosial Pemerintah Kota
the paper disc after submerging into this oil, onto agar Bogor. Bogor, 4 November 2004.
media grown with strain of Salmonella, indicating that Soeka, Y.S., J. Sulistyo, dan E. Naiola. 2008. Analisis biokimia
this oil exhibited activity against the growth of the minyak kelapa hasil ekstraksi secara fermentasi. Biodiversitas
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