Antibacterial and antioxidant activities of endophytic fungi extracts of medicinal plants from Central Sulawesi

Abstract

Endophytic fungi are a group of fungi which grow inside the plant tissues without causing negative symptoms to the host plant and are able to produce biologically active substances. This research was carried out to evaluate the antibacterial and antioxidant activity of 40 endophytic fungi isolated from 10 species of medicinal plants collected from Palolo, Central Sulawesi. Thin layer chromatography (TLC) bioautography guided screenings were done to evaluate antibacterial and antioxidant activities. The antibacterial activity was done against Staphylococcus aureus InaCC-B5 and Escherichia coli InaCC-B4, while antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Minimum inhibitory concentrations (MICs) of active extracts were further evaluated against these bacteria, while half-maximal inhibitory concentration (IC 50) of active extracts was determined by the microdilution broth method. The results of TLC bioautography screening showed 30 extracts inhibited the growth of S. aureus, 29 extracts inhibited the growth of E. coli, 27 extracts inhibited both S. aureus and E. coli, and 23 extracts posessed antioxidant activity. There were six extracts with MIC value of <100 µg/ml against S. aureus and nine extracts with MIC value of <100 µg/ml against E. coli. Six extracts indicated very strong antioxidant activity.

Full text

Journal of Applied Pharmaceutical Science Vol. 8(08), pp 069-074, August, 2018
Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2018.8811
ISSN 2231-3354
© 2018 Praptiwi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-
ShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
*Corresponding Author
Andria Agusta; Natural Product Chemistry Laboratory, Reseach Center
for Biology, Indonesian Institute of Sciences. Jl. Raya Bogor Km. 46,
Cibinong 16911, Indonesia. E-mail: andr002 @ lipi.go.id
Antibacterial and antioxidant activities of endophytic fungi extracts
of medicinal plants from Central Sulawesi
Praptiwi, Marlin Raunsai, Dewi Wulansari, Ahmad Fathoni, Andria Agusta*
Natural Product Chemistry Laboratory, Reseach Center for Biology, Indonesian Institute of Sciences, Jl. Raya Bogor Km. 46, Cibinong 16911, Indonesia.
ARTICLE INFO ABSTRACT
Article history:
Received on: 09/12/2017
Accepted on: 08/02/2018
Available online: 31/08/2018
Endophytic fungi are a group of fungi which grow inside the plant tissues without causing negative symptoms to
the host plant and are able to produce biologically active substances. This research was carried out to evaluate the
antibacterial and antioxidant activity of 40 endophytic fungi isolated from 10 species of medicinal plants collected
from Palolo, Central Sulawesi. Thin layer chromatography (TLC) bioautography guided screenings were done to
evaluate antibacterial and antioxidant activities. The antibacterial activity was done against Staphylococcus aureus
InaCC-B5 and Escherichia coli InaCC-B4, while antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl
(DPPH) method. Minimum inhibitory concentrations (MICs) of active extracts were further evaluated against these
bacteria, while half-maximal inhibitory concentration (IC50) of active extracts was determined by the microdilution
broth method. The results of TLC bioautography screening showed 30 extracts inhibited the growth of S. aureus,
29 extracts inhibited the growth of E. coli, 27 extracts inhibited both S. aureus and E. coli, and 23 extracts posessed
antioxidant activity. There were six extracts with MIC value of <100 µg/ml against S. aureus and nine extracts with
MIC value of <100 µg/ml against E. coli. Six extracts indicated very strong antioxidant activity.
Key words:
antibacterial, antioxidant,
endophytic fungi,
bioautography.
INTRODUCTION
Endophytic fungi are groups of fungi with very
specic ecosystem inside plant tissues and produces varieties of
secondary metabolites (Agusta, 2009). Secondary metabolites
from endophytic fungi show important biological activities
such as antioxidant, anticancer, immunomodulatory, antivirus,
antituberculosis, anti-parasite and insecticides (Hussain et al.,
2014). Endophytic fungi produce secondary metabolites similar to
the host plant; therefore, endophytic fungi can be used as a source
of producing active metabolites and leads in drug developments
(Strobel, 2003; Owen and Hundley, 2004). The study on the
isolation and evaluation of bioactivities of endophytic fungi from
medicinal plants are increasing lately.
On the other hand, antimicrobial resistance has been
a major concern in the health care system globally (Ferri et
al., 2017). Discovery of novel and active metabolites against
pathogenic microbes as well as to overcome antimicrobial
resistance become very important. In addition to health problems
with increasing resistance, there is also a growing tendency to
search natural antioxidants to overcome degenerative disease
problems. Reactive oxygen species (ROS) are by-products of
biological reactions that cause oxidative damage to biomolecules
and play vital roles in programmed cell death (Cui et al., 2015).
To overcome the negative effect of excessive ROS in human body,
exogenous antioxidant is required. The main characteristic of
antioxidant compounds is the ability to capture and stabilize free
radicals (Prakash et al., 2011), inhibit or delay the occurrence of
free radical reactions due to the presence of relative oxygen; these
properties become important in the prevention of various diseases,
such as cancer and coronary heart disease (Leong and Shui, 2002).
Medicinal plants are reported as host of some endophytic
fungi that are involved in the co-production of active metabolites
(Alvin et al., 2014). The study conducted by Ilyas (2009); Praptiwi
et al. (2010); Praptiwi et al. (2015) showed that endophytic fungi
isolated from medicinal plants such as gambier (Uncaria gambier),
cinnamon (Cinnamomum burmannii) dan Zingiberaceous plants
have antioxidant and/or antibacterial activity.

Praptiwi et al. / Journal of Applied Pharmaceutical Science 8 (08); 2018: 069-074070
Khiralla et al. (2015) has identied and classied 21
endophytic fungi from ve medicinal plants of Sudan origin
and some contain phenol compounds that have the potential as
antioxidant natural sources. The present study aims to isolate and
evaluate the antibacterial and antioxidant activity of endophytic
fungi from ten species of medicinal plants originating from Palolo.
MATERIAL AND METHODS
Material
10 species of plants belonging to seven families which
are Urticaceae (Villebrunea rebescens (Blume), Poikilospermum
suaveolens (Blume) Merr.), Euphorbiaceae (Euphorbia
heterophylla L., Acalypha caturus Blume), Asteraceae (Blumea
balsamifera (L.) DC.), Zingiberaceae, Piperaceae (Piper peltatum
L.), Lamiaceae (Plectranthus scutellarioides (L.) R. Br), and
Verbenaceae (Cleodendron fragrans Wild.) were collected from
Palolo, Central Sulawesi. Identication of the plant specimens
were done at Herbarium Bogoriense, Research Center for Biology-
Indonesian Institute of Sciences.
Isolation of endophytic fungi
Leaves, stems, and rhizomes collected from the eld
were stored at low temperature. After arriving in the laboratory,
these samples were cleaned under tap water and immersed in 70%
ethanol for 1 minutes, then immersed in 5.3% Na-hypochlorite for
5 minutes and nally immersed in 70% ethanol for 30 seconds.
Samples were dried under aseptic conditions. The sterilized
samples were cut aseptically into small pieces (1 × 1 cm2), and
then, placed on top of the Corn Meal Malt Agar (CMMA) growth
medium added with chloramphenicol 0.05 mg/ml, and incubated
at room temperature for 1 week. The emerging colonies were
subcultured several times on Potato dextrose agar (PDA) to obtain
pure isolates.
Secondary metabolites extraction from endophytic fungi
Pure isolate of endophytic fungi was cultured on broth
medium [Potato dextrose broth (PDB)] (200 ml) and incubated in
dark condition, at room temperature for 3 weeks. After incubation
period is completed, growth media and endophytic fungi biomass
were extracted three times with ethyl acetate. The extract was
evaporated by rotary evaporation and the concentrated extract was
stored in the glass vial.
Chemical compounds analysis by Thin Layer
Chromatography (TLC)
The analysis of chemical compounds of endophytic fungi
extracts were performed on silica gel thin layer chromatography
(TLC) plates (silica gel GF254, Merck). The dried extract was
prepared in 10 mg/ml. 10 µl of extract was transferred on TLC
plate and developed in CH2Cl2:MeOH (10:1). Separated chemical
compounds were visualized under 254 nm and 366 nm ultraviolet
(UV) light followed by spraying with spray reagent 1% Ce(SO4)2
and 1% vanillin sulphuric acid.
Detection of antibacterial activity by TLC-bioautography
TLC-bioautography guided screening was performed to
evaluate the antibacterial potency of endophytic fungal extracts.
10 µl of extract was transferred on TLC plate and dried. Plate was
then dipped into bacterial suspension, followed by incubating the
plate under humid condition for 18 hours at 37°C. After incubation
was completed, plates were sprayed with iodonitrotetrazolium
p-violet (INT, Sigma). Growth inhibition of bacteria was observed
by clear zone formation around the extract. The active extracts
were further analyzed by developing the extract with mobile
phase CH2Cl2:MeOH (10:1). The plate was dried and sprayed with
iodonitrotetrazolium p-violet (INT, Sigma).
Detection of antioxidant activity by TLC-bioautography
10 µl of extract was transferred on TLC plate and
catechin used as positive control was also transferred on the
TLC plate. The plate was dried and sprayed with 0.02% DPPH
in methanol. Yellow spot on purple background indicated the
antioxidant activity. The active extract was developed with
CH2Cl2:MeOH (10:1). After drying, plate was sprayed with 0.02%
DPPH in MeOH.
Determination of minimum inhibitory concentration
The minimum inhibitory concentration (MIC) of active
extracts were determined by serial microdilution in 96-well
microplate (Pessini et al., 2003). The wells in column A were
lled with 100 μl of Mueller-Hinton Broth (MHB) medium and
100 µl stock solution of extract (1024 µg/ml) and homogenized.
Columns B through H were lled with 100 µl of MHB. A serial
dilution was carried out with corresponding nal concentration
(256 µg/ml). The test was done in triplicate. After the dilution
process, each well was added with 100 μl of bacterial suspension
(106 cfu/ml). In similar way, it was done with the positive control
of chloramphenicol, growth media as negative control. Microtiter
plate sealed with paralm and incubated at 37°C for 24 hours.
After incubation was completed, each well was added with 10
µl INT 4 mg/ml. The MIC was the lowest concentration showing
clear wells that indicate the absence of bacterial growth.
Determination of IC50 of active extract
The IC50 of the extract was determined by serial
microdilution in 96-well microplate by Takao et al. (2015) with
minor modication. The wells in column A were lled with 195 μl
MeOH and 5 µl extract (10240 µg/ml) and homogenized. Columns
B through H were lled with 100 µl of MeOH. A serial dilution
was carried out with corresponding concentration (256 µg/ml).
After the dilution process completed, each well was added with
100 μl of DPPH (61.50 µg/ml). Methanol was used as negative
control, while catechin was used as positive control. Microplates
were incubated in dark condition at room temperature for 90
minutes. The absorbances of the samples were measured at 517
nm. Antioxidant activity index (AAI) was calculated as follows:
AAI = nal concentration of DPPH in the reaction/IC50
IC50: the concentration of 50% inhibition was calculated by linear
regression equation.
RESULT AND DISCUSSION
A total of 40 isolates of endophytic fungi recovered
from 10 species of medicinal plants collected from Palolo, Central
Sulawesi. The results in Table 1 show that a plant part is colonized

Praptiwi et al. / Journal of Applied Pharmaceutical Science 8 (08); 2018: 069-074 071
by more than one endophytic fungus and different plant parts
might have different composition of endophytic fungi community.
This nding is in accordance with Zabalgogeazcoa (2008) that one
species of plant inhabited by more than one endophytic fungus.
Previous reports by Huang et al. (2008) and Ilyas (2009) showed
similar results that one species of plant inhabited by various
endophytic fungi. The distribution, composition, and population
structure of endophytic fungi rely largely on the taxonomy,
genetic background, age, and tissues of the host plants, and the
types of environments (Sieber, 2007; Jia et al., 2016). Older plant
parts may be colonized by higher number of endophytes than the
younger ones (Zabalgogeazcoa, 2008).
Table 1: MIC of endophytic fungi extracts isolated from plants collected from Palolo, Central Sulawesi.
No Sample
MIC (ug/ml)
No Sample
MIC (ug/ml)
S. aureus E. coli S. aureus E. coli
1 PAL-01B1 64 64 21 PAL-09D2 NT 256
2 PAL-01B2 32 32 22 PAL-10B1 128 256
3 PAL-01D1 NT NT 23 PAL-10B2 256 128
4 PAL-01D2 NT 256 24 PAL-10B3 128 8
5 PAL-02D1 128 256 25 PAL-10D1 256 64
6 PAL-02D2 256 NT 26 PAL-10D3 256 128
7 PAL-03B1 64 32 27 PAL-10D8 NT NT
8 PAL-03B2 256 128 28 PAL-11B1 NT 256
9 PAL-03B3 128 256 29 PAL-11B2 NT NT
10 PAL-03D1 256 64 30 PAL-11B3 128 NT
11 PAL-03D2 256 256 31 PAL-11D1 64 256
12 PAL-04R1 256 256 32 PAL-11D2 256 256
13 PAL-04R2 128 128 33 PAL-14D1 64 128
14 PAL-07B1 8 8 34 PAL-14D2 128 8
15 PAL-07B2 128 64 35 PAL-14D3 256 64
16 PAL-07D1 256 128 36 PAL-15B1 256 128
17 PAL-09B1 256 NT 37 PAL-15B2 NT NT
18 PAL-09B2 128 256 38 PAL-15D1 NT 256
19 PAL-09B3 NT NT 39 PAL-15D2 NT NT
20 PAL-09D1 NT NT 40 PAL-15D3 NT NT
Analysis of the chemical compounds of the extract
The chemical compounds of the endophytic fungi were
analyzed by TLC in order to separate the chemical compounds
within the extract. TLC is an important method for qualitative
and quantitative analysis of drugs and has several advantages
compared to HPLC and GC methods (Pyka, 2014). The plates
were sprayed with color reagents (vanillin reagent and cerium
reagent) to detect compounds in extract.
Observations under 254 nm showed different chemical
compounds in each extract which emitted green and dark-colored
compounds. A substance having a maximum wavelength (λmax) of
250–260 nm may contain aromatic groups such as aromatic amino
acid, simple phenol, and purines or pyrimidines (Harborne, 1973).
Observation under 366 nm showed that TLC plates as background
emitted purple and spot dark-colored chemical compounds.
Substances having λmax 200–400 nm indicated the presence of
compounds has an aromatic group or a conjugated double bond
(Fried and Sherma, 1999). TLC plates sprayed with vanillin and
cerium showed the presence of different chemical compounds in
extract that was characterized by stain spots with multiple colors.
Crude extract of endophyic fungi (Figure 1) contained several
chemical compounds indicated by several spots with different Rf.
These chemical compounds might have biological activities and
mixture of chemical compounds in crude extract may increase the
potential of the active component to produce additive or synergistic
effects, while others may be neutral or inhibit (Dhankhar et al.,
2012).
Antibacterial activity detection by TLC
Antibacterial screening of endophytic fungi extracts
was done by TLC method. TLC method is nest, rapid, efcient,
and uncomplicated method (Masoko and Ellof, 2006; Shahverdi
et al., 2007), requires small amount of test sample and simple
interpretation of results (Valle Jr. et al., 2016). TLC dot-blot direct
bioautography of antibacterial activity screening shown in Figure
2. The results of antibacterial screening of 40 endophytic fungi
extracts showed that 30 extracts inhibited the growth of S. aureus,
29 extracts inhibited the growth of E. coli and 27 extracts were
able to inhibit S. aureus as well as E. coli. The growth inhibition
of bacteria was indicated by clear zone formation on TLC against
a purple background (Das et al., 2010). The purple colour on TLC
plate after spraying with INT was resulted from the conversion of
INT to intensely colored formazan by the dehydrogenases enzyme
of living microorganisms (Silva et al., 2005). Shahverdi et al.
(2007) also stated that INT interacted with viable microorganisms
caused a colour change of INT to red-purple one. Screening
antibacterial activity by TLC dot-blot is simple and time saving
method; however, the component mixtures in the crude extract

Praptiwi et al. / Journal of Applied Pharmaceutical Science 8 (08); 2018: 069-074072
can have synergistic or antagonistic effects (Choma and Jesionek,
2015). The active extracts developed with mobile phase to separate
the bioactive compounds in the extract (Figure 3). Separated
bioactive compounds with antibacterial properties indicated by
white band formation.


Fig. 1: Chromatograms of endophytic fungal extracts developed in dichloromethane-methanol (10:1 v/v), (a) viewed under 254 nm wavelength, (b) viewed under 366
nm wavelength, (c) sprayed with vanillin reagent, (d) sprayed with cerium reagent.
Fig. 2: TLC dot-blot assay for antibacterial activity of endophytic fungi against
E. coli (top) and S. aureus (bottom).
Fig. 3: Bioautograms of endophytic fungi against E. coli (top) and S. aureus
(bottom). The TLC plates were developed in dichloromethane:methanol (10:1
v/v). Clear bands indicated antibacterial activity.
The minimum inhibitory concentration (MIC) of active
extracts was assessed against E. coli and S. aureus. The result in
Table 1 showed that MIC values of PAL endophytic fungi extracts
ranging from 8 to 256 µg/ml. MIC in the range of 100–1000 µg/
ml could be classied as antimicrobial (Borges et al., 2012). The
MIC of several endophytic fungi were <100 µg/ml. According to
Pessini et al. (2003), the MIC value of extract <100 µg/ml was
classsied as good antibacterial activity, while extracts with MIC
value ranging from 100 µg/ml to 500 µg/ml classied as moderate
activity. Among 40 endophytic fungi tested for antibacterial
activity, endophytic fungus PAL-07B1 derived from Piper
peltatum, has good antibacterial activity against S. aureus and
E. coli, with equivalent MIC for both isolates (8 µg/ml). Several
previous studies reported the antibacterial activity of ethyl acetate
extracts of endophytic fungi isolated from Piper (Orlandelli et al.,
2012; Astuti et al., 2014). This result suggested the endophytic
fungus PAL-07-B1 contains potential bioactive compounds as
antibacterial.
Antioxidant activity by TLC-bioautography
The antioxidant activity of extracts was done by DPPH
free radical scavenging activity. DPPH radical when receiving an
electron from antioxidant compound would be reduced to DPPH.
The violet color of DPPH radical turned into yellow (Pavithra and
Vadivukkarasi, 2015). Screening for antioxidant activity by dot-
blot TLC (Figure 4) showed 23 extracts had antioxidant activity.
Antioxidant activity is indicated by the color change to yellow
against a purple background (Dewanjee et al., 2015). The intensity
of yellow color indicates the antioxidant capacity. Further analysis
of active extract was shown in Figure 5.
The developed TLC-bioautography of endophytic fungi
extracts (Figure 5) showed several compounds possess antioxidant
activity within extract. This is indicated by the formation of

Praptiwi et al. / Journal of Applied Pharmaceutical Science 8 (08); 2018: 069-074 073
yellowish white bands. Further analysis of active antioxidant
extracts is to determine its IC50 value (Table 2).
Fig. 4: TLC dot-blot assay for antioxidant activity of endophytic fungi extracts
isolated from medicinal plants from Central Sulawesi.

Fig. 5: TLC-bioautogram of antioxidant activity of endophytic fungi extracts.
The yellowish white band indicates the compounds with antioxidant activity.
Table 2: IC50 and antioxidant activity index (AAI) of endophytic fungi extracts.
No Sample IC50 (ug/ml) AAI value Criteria of AAI value
1 PAL-01B2 5.26 5.846 Very strong
2 PAL-01D2 7.84 3.922 Very strong
3 PAL-02D1 26.00 1.183 Strong
4 PAL-03D1 107.51 0.286 Moderate
5 PAL-04R1 52.70 0.583 Moderate
6 PAL-04R2 10.02 3.069 Very strong
7 PAL-07B2 77.60 0.396 Moderate
8 PAL-09B1 99.72 0.308 Moderate
9 PAL-11B1 10.03 3.066 Very strong
10 PAL-14D3 14.06 2.187 Very strong
11 PAL-15D1 43.15 0.713 Moderate
12 Catechin 1.71 17.982 Very strong
Based on the criteria of AAI value by Scherer and Godoy
(2009), there are ve extracts that displayed very strong antioxidant
activity (PAL 01-B2, PAL 01-D2 from Villebrunearubescens
(Urticaceae), PAL 04-R2 (Zingiberaceae), PAL 11-B1 from
Clerodendronfragrans (Verbenaceae), and PAL 14-D3 from
Acalyphacaturus (Euphorbiaceae); one extract displayed strong
antioxidant activity and ve extracts displayed moderate activity.
Previous study by Praptiwi et al. (2016) showed some endophytic
fungi isolated from Zingiberaceae had strong antioxidant activity,
while endophytic fungi from other studied plants had no previous
reports. The very strong antioxidant activity of extracts was related
to many bioactive compounds as antioxidant within extract may
act synergistically.
CONCLUSION
This study revealed the presence of bioactive secondary
metabolites produced by endophytic fungi from several medicinal
plants collected from Central Sulawesi with antibacterial and/or
antioxidant activity. Endophytic fungi PAL-01B2 and PAL-07B1
showed good antibacterial activity against S. aureus InaCC-B5
and E. coli InaCC-B4. Five endophytic fungi showed very strong
radical scavenging activity (PAL 01-B2, PAL 01-D2, PAL 04-R2,
PAL 11-B1), and PAL 14-D3. Further studies needed to isolate and
purify bioactive compounds, which is responsible for antibacterial
and antioxidant activity. These ndings indicated that endophytic
fungi from medicinal plants collected from Palolo, Central
Sulawesi could be potential for the development as pharmaceutical
agents.
ACKNOWLEDGEMENT
The authors are thankful to Research Center for Biology,
Indonesian Institute of Sciences for nancial support through
DIPA-Pusat PenelitianBiologi.
REFERENCES
Agusta A. 2009. Biologi dan Kimia Jamur Endot. Bandung,
Indonesia. InstitutTeknologi Bandung Publisher.
Alvin A, Miller KI, Neilan BA. Exploring the potential
of endophytes from medicinal plants as sources of antimycobacterial
compounds. Microbiol Res., 2014; 169(7-8):483-95. doi.org/10.1016/j.
micres.2013.12.009.
Astuti P, Wahyono, Nababan OA. Antimicrobial and cytotoxic
activities of endophytic fungi isolated from Piper crocatum Ruiz & Pav.
Asian Pacic Journal of Tropical Biomedicine, 2014, Volume 4, Supplement
2 Pages S592-S596.
Borges, A.; Saavedra, M.J.; Simões, M. The activity of ferulic
and gallic acids in biolm prevention and control of pathogenic bacteria.
Biofouling, 2012, 28, 755–767.
Choma IM and Jesionek W. TLC-Direct Bioautography as
a High Throughput Method for Detection of Antimicrobials in Plants.
Chromatography, 2015, 2, 225-238; doi:10.3390/chromatography2020225.
Cui J-L, Guo T-T, Ren Z-X, Zhang N-S, Wang M-L. Diversity
and antioxidant activity of culturable endophytic fungi from Alpine plants
of Rhodolacrenulata, R. angusta and R. schalinensin. PLoS ONE, 2015;
10(3): e0118204. doi:10.1371/journal. pone.0118204.
Das K, Tiwari RKS, Shrivastava DK. Techniques for evaluation
of medicinal plant products as antimicrobial agent: current methods and
future trends.J. Med Plants Res, 2010; 4(2): 104-111. doi: 10.5897/
JMPR09.030.
Dhankhar S, Kumar S, Dhankhar S, Yadav JP. Antioxidant
activity of fungal endophytes isolated from Salvadora oleoides Decne. Int J
Pharm and Pharm Sci, 2012; 4 (2): 380–85.
Dewanjee S, Gangopadhyay M, Bhattacharyaa N, Khanra R, Dua
TK. Bioautography and its scope in the eld of natural product chemistry.
Journal of Pharmaceutical Analysis, 2015; 5(2): 75-84.
Ferri M, Ranucci E, Romagnoli P, Giaccone V. Antimicrobial
resistance: A global emerging threat to public health systems. Crit Rev Food
Sci Nutr, 2017; 57(13):2857-2876. doi: 10.1080/10408398.2015.1077192.
Fried B, Sherma J. 1999. Thin-Layer Chromatography 4th ed,
revised and expanded. New York, USA. Marcel Dekker, Inc.
Harborne JB. 1973. Phytochemical methods A guide to modern
techniques of plantanalysis 3rd Ed. London Chapman & Hall.
Huang W-Y, Cai Y-Z, Hyde KD, Corke H, Sun M. Biodiversity

Praptiwi et al. / Journal of Applied Pharmaceutical Science 8 (08); 2018: 069-074074
of endophytic fungi associated with 29 traditional chinese medicinal plants.
Fungal Diversity, 2008; 33: 61–75.
Hussain H, Kliche-Spory C, Al-Harrasi A, Al-Rawahi A, Abbas
G, Green IR, Schulz B, Krohn K, Shah A. Antimicrobial constituents from
three endophytic fungi. Asian Pac J Trop Med, 2014; 7 (S1): S224–27.
doi:10.1016/S1995-7645(14)60236-4.
Ilyas M. Biodiversity of endophytic fungi associated with
Uncaria Gambier Roxb. (Rubiaceae) from West Sumatra. Biodiversitas,
Journal of Biological Diversity, 2009; 10 (1): 23–28. doi:10.13057/biodiv/
d100105.
Jia M, Chen L, Xin H-L, Zheng C-J, Rahman K, Han T and Qin
L-P. A Friendly Relationship between Endophytic Fungi and Medicinal
Plants: A Systematic Review. Front. Microbiol., 2016; 7:906. doi: 10.3389/
fmicb.2016.00906.
Khiralla A, Mohamed I, Thomas J, Mignard B, Spina R, Yagi S,
Laurain-Mattar D. 2015. A pilot study of antioxidant potential of endophytic
fungi from some Sudanese medicinal plants. Asian Pac J Trop Med, 2015; 8
(9): 701–4. doi:10.1016/j.apjtm.2015.07.032.
Leong LP, Shui G. An investigation of antioxidant capacity
of fruits in Singapore markets. Food Chemistry, 2002; 76 (1): 69–75.
doi:10.1016/S0308-8146(01)00251-5.
Masoko P and Eloff JN: Bioautography indicates the multiplicity
of antifungal compounds from twenty-four southern African Combretum
species (Combretaceae). Afr J Biotechnol, 2006; 5:1625–1647.
Orlandelli, RC, Alberto RN, Almeida TT, Azevedo JL, Pamphile
JA. In vitro Antibacterial Activity of Crude Extracts Produced by Endophytic
Fungi Isolated from Piper hispidum. Journal of Applied Pharmaceutical
Science, 2012; Vol. 2 (10), pp. 137-141. DOI: 10.7324/JAPS.2012.21027.
ISSN 2231-3354.
Owen NL, Hundley N. Endophytes--the chemical synthesizers
inside plants. Sci Prog., 2004; 87(Pt 2):79-99.
Pavithra K, Vadivukkarasi S. Evaluation of free radical
scavenging activity of various extracts of leaves from Kedrostis foetidissima
(Jacq.) Cogn. Food Science and HumanWellness, 2015; 4: 42–46.
Pessini GL, Dias-Filho BP, Nakamura CV, Cortez DAG.
Antibacterial activity of extracts and neolignans of Piper regnellii (Miq.)
C.DC. var pallescens (C.DC) Yunck. Mem. I. Oswalso Cruz 98, 1115-1120.
Prakash A, Rigelhof F, Miller E. Antioxidant activity. Eur Rev
Med Pharmacol Sci, 2011;15 (4): 376–78. doi:10.1016/j.bmcl.2010.12.025.
Praptiwi, Jamal Y, Fathoni A, and Agusta A. Antimicrobial
metabolite from the culture of endophytic fungus AFK-8 isolated from
kayu kuning (Archangelisia ava (L.) Merr.) Proceeding of International
Seminar: Biotechnology for Enhancement the Tropical Biodiversity, 2010;
pp. 64-72.
Praptiwi, Ilyas M, Fathoni A, Wulansari D, and Agusta A.
Antibacterial screening of the culture of endophytic fungal extracts isolated
from cinnamon stick (Cinnamomum burmannii (Nees&T.Nees) BLUME)
Teknologi Indonesia, 2015; 38 (1): 33–41.
Pyka A. Review: Detection progress of selected progress in TLC.
BioMed Research International, 2014; Volume 2014, Article ID 732078,
http://dx.doi.org/10.1155/2014/732078.
Scherer R, Godoy HT. Antioxidant activity index (AAI) by the
2,2-diphenyl-1-picrylhydrazyl method. Food Chemistry, 2009; vol. 112 (3):
654-658.
Shahverdi AR, Abdolpour F, Monsef-Esfahani HR, Farsam H.
A TLC bioautographic assayfor the detection of nitrofurantoin resistance
reversal compound. J Chromatogr B AnalytTechnol Biomed Life Sci.,
2007;850(1-2):528-30.
Sieber TN. Endophytic fungi in forest trees: are they mutualists?
Fung. Biol. Rev., 2007:21:75–89.2007. doi:10.1016/j.fbr.2007.05.004.
Accessed: January 4, 2018.
Silva MTG, Simas SM, Batista TGFM. Studies on antimicrobial
activity, in vitro, of Physalis angulata L. (Solanaceae) fraction and physalin
B bringing out the importance of assay determination. Mem. Instit.
OswaldoCruz, 2005;100:779–782.
Strobel GA. Endophytes as sources of bioactive products.
Microbes Infect., 2003; 5(6):535-44.
Takao LK., Imatomi M., and Gualtieri. Antioxidant activity
and phenolic content of leaf infusions of Myrtaceae species from Cerrado
(Brazilian savanna). Braz. J. Biol, 2014;75(4): 948-952.
Valle Jr. DL, Puzon JJM, Cabrera EC, andRivera WL. Thin
Layer Chromatography-Bioautography and Gas Chromatography-Mass
Spectrometry of Antimicrobial Leaf Extracts from Philippine Piper betle
L. against Multidrug-Resistant Bacteria Evidence-Based Complementary
and Alternative Medicine, 2016; Volume 2016, Article ID 4976791, http://
dx.doi.org/10.1155/2016/4976791.
Zabalgogeazcoa I. Review. Fungal endophytes and their
interaction with plant pathogens. Spanish Journal of Agricultural Research,
2008; 6 (Special issue), 138-146. Available online at www.inia.es/sjar
ISSN: 1695-971-X, Accessed: August 10, 2017.
How to cite this article:
Praptiwi, Raunsai M, Wulansari D, Fathoni A, Agusta A.
Antibacterial and antioxidant activities of endophytic fungi
extracts from medicinal plant from Central Sulawesi. J App
Pharm Sci, 2018; 8(08): 069-074.