BIOACTIVE COMPOUND ANALYSIS AND BIOACTIVITIES OF ENDOPHYTIC BACTERIA FROM CISSUS QUADRANGULARIS
HTML Full TextBIOACTIVE COMPOUND ANALYSIS AND BIOACTIVITIES OF ENDOPHYTIC BACTERIA FROM CISSUS QUADRANGULARIS
Parvin Zeenath Anwar 1, Uma Gowrie Sezhian * 1 and Arunagirinathan Narasingam 2
Department of Plant Biology and Plant Biotechnology 1, Ethiraj College for Women (Autonomous), Chennai - 600008, Tamil Nadu, India.
Department of Microbiology & Biotechnology 2, Presidency College, Chennai - 600005, Tamil Nadu, India.
ABSTRACT: Endophytic bacteria were isolated from the stem of Cissus quadrangularis on Nutrient agar medium. The bacterial isolates were cultured in Nutrient broth. Ethyl acetate extracts of endophytic bacteria were evaluated for the presence of bioactive compounds and total phenolic content. The phytochemical screening of extracellular ethyl acetate extracts showed the presence of phenols. The total phenol content in the extracellular extract of a bacterium labeled as CqB9 was found to be 7.05 ± 0.13 mg/g. The extracellular crude extract of CqB9 displayed considerable antibacterial activities against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Klebsiella pneumoniae. GC-MS analysis of CqB9 extract revealed several peaks indicating the presence of different secondary metabolites having bioactive potential. The extract of CqB9 was found to possess antioxidant activity with an IC50 value of 157.5 ± 0.4 µg. The cytotoxic activity using MTT assay showed that the crude extract inhibited the proliferation of the MCF-7 cell line in a dose-dependent manner. CqB9 was found to be similar to Achromobacter anxifer by 16S rRNA sequencing. The bioactive components such as phenols and those predicted by GC-MS may contribute to the bioactivities of the isolate. The results reveal that the endophytic bacterium isolated from Cissus quadrangularis has the potential to produce bioactive compounds. The endophytic bacteria and the metabolites produced by them need to be explored further for potential source and novel natural bioactive compounds.
Keywords: |
Endophyte, Cissus quadrangularis, Antimicrobial, Antioxidant, GC-MS, Cytotoxic
INTRODUCTION: There is an emerging need for bioactive compounds derived from natural sources for the development of drugs 1. Endophytes are a specific group of microorganisms that are found in internal tissues of healthy plant and do not cause visible damage to their hosts.
Exploring the untapped natural products from the endophytes increases the chances of finding novel compounds. Attention has been directed towards endophytes of medicinal plants as these plants are being traditionally used for generations to treat symptoms of numerous diseases.
Exploring bioactive compounds of endophytic bacteria isolated from an ethnomedicinal plant may provide an alternative source for the extraction of potential metabolites on a large scale 2. Cissus quadrangularis is a perennial plant of the family Vitaceae. It is also known as an adamant creeper, square stalked vine, veldt grape, devil's backbone, asthisamharaka, hadjod, pirandai and mangara valli 3. Cissus quadrangularis has been investigated scientifically in animal models based on the folk and traditional uses to validate its potential to cure a variety of ailments 4. Cissus quadrangularis has potent fracture healing property and also possesses antimicrobial, antiulcer, antioxidative, antiosteo-porotic and gastroprotective activities 5. Stem and root extracts of this plant are important in the management of various ailments. Some other reports on Cissus quadrangularis justify its effectiveness in antioxidant and free radical scavenging activity in-vitro 6,7. Cissus quadrangularis also has anticancerous potential 8, 9.
Most of the literature covers work on bioactivities of endophytic fungi, but endophytic bacteria remain unexplored. Meagre reports are available on the endophytic bacteria of Cissus quadrangularis and their bioactivities. As a part of our ongoing efforts towards finding novel bioactive agents from natural resources, we investigated bioactive components, antibacterial, antioxidant, and cyto-toxic efficacy of endophytic bacterium isolated from the stem of Cissus quadrangularis.
MATERIALS AND METHODS:
Isolation of Endophytic Bacteria from Stem of Cissus quadrangularis: The stems of plant Cissus quadrangularis were collected from the Department of Plant Biology and Plant Biotechnology, Ethiraj College for Women, Chennai, Tamil Nadu. The plant sample was identified and authenticated at Botanical Survey of India (BSI) Coimbatore, Tamil Nadu (Confirmation I.D No: BSI/SRC/5/23/2018/Tech/1884). Fresh and healthy stems were washed thoroughly under running tap water and were surface sterilized. Finally, they were washed thrice with sterile distilled water. Thereafter, the outer part of the stem was excised off and the remaining part of the stem was sliced into thin sections. The sections of stems were placed over the Nutrient Agar plate and incubated for 2-4 days. Aseptic conditions were maintained throughout the steps. The endophytic bacteria emerging from the sections of stem were isolated and purified by continuous subculturing 10.
Preparation of Crude Extract of Endophytic Bacteria: Each bacterial culture was inoculated in Nutrient Broth and incubated for 10 days. Supernatants were collected after centrifugation. The supernatants were subjected to solvent extraction using ethyl acetate. The concentrated residue of each extract was regarded as a crude sample and stored under refrigeration for further use.
Analysis of Bioactive Compounds: The crude extracts of the endophytic bacteria were subjected to phytochemical analysis for the presence of secondary metabolites, namely carbohydrates, tannins, phenols, flavonoids, and terpenoids by the standard methods 11, 12.
Determination of Total Phenolic Content: The amount of phenol in each of the extracellular extracts was determined by the Folin-Ciocalteu’s colorimetric method and calculated from a calibration curve obtained with gallic acid as standard (10 mg/10 ml). From the standard solution, 20 to 100 µl was taken and added to different test tubes. The extract was added in a separate test tube at a concentration of 10 mg/ml and 5ml of Folin-Ciocalteu’s reagent (1:10 dilution) was added, and the contents were mixed thoroughly. 4ml of 0.7 M sodium carbonate was added, and the mixture was incubated for 30 min. The absorbance was measured at 765 nm in a UV-Visible spectrophotometer. The results were expressed in gallic acid equivalence of the samples (GE) mg/g of the extract 13.
Antimicrobial Activity: The crude extracts of the endophytic bacteria were screened for antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Klebsiella pneumoniae. This assay was carried out using standard agar well diffusion method 14. Nutrient agar medium was used for the assay. The plates were incubated for 24 h at 37 °C. Zones of inhibition were observed and measured.
Identification of Bioactive Compounds using GC-MS Analysis: The extracellular extract of CqB9 was subjected to Gas Chromatography-Mass Spectrometry (GC-MS) to analyze the bioactive compounds. GC-MS analysis was performed at the SAIF, IIT-Madras, Chennai, Tamil Nadu. The endophytic extract was subjected to GC and MS, JOEL GC mate equipped with secondary electron multiplier (Agilent Technologies 6890N Network GC system for Gas chromatography). The column (HP5) was fused with silica 50 m × 0.25 mm I.D. The experimental conditions were maintained at 20 min at 100 °C, column temperature was 235 °C for 3 min; injector temperature was 240 °C; carrier gas: helium; and split ratio was 5:4. The sample (1μl) was evaporated in a splitless injector at 300 °C, and the run time was 30 min. The components were identified by gas chromatography coupled with mass spectrometry. The spectrum of GC-MS was analyzed using the NIST library. The name, molecular weight and molecular formula of the compounds were ascertained.
DPPH Free Radical Scavenging Activity: The ability of the samples to reduce the DPPH radical (1, 1-diphenyl-2-picrylhydrazyl) was investigated by the method described by Blois 15. A stock solution of the compound was prepared for the concentration of 10 mg/ml. Different concentrations of the extract (200, 400, 600, 800, 1000 µg) were added at an equal volume to the methanolic solution of DPPH (0.1 mM). The reaction mixture was incubated for 30 min at room temperature; the absorbance was recorded at 517 nm. The experiment was repeated three times. Ascorbic acid was used as a standard control. The annihilation activity of free radicals was calculated in percent inhibition according to the formula
% Inhibition = {(A of control – A of Test) / A of control} × 100
Cytotoxic Activity: Breast cancer cell line MCF-7, obtained from National Centre for Cell Science (NCCS), Pune, India, was used to determine the cytotoxic activity. The cells were maintained in Minimal Essential Media supplemented with 10% FBS, penicillin in 5 % CO2 at 37 ºC. Cytotoxicity of the extract at various concentrations was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, as described by Mosmann 16 with modification. The viable cells were determined by the absorbance at 570 nm by a microplate reader. All readings were taken in triplicates.
% Inhibition = (Control Test / Control) × 100
16S rRNA Sequencing: The endophytic bacterium CqB9 was identified by 16S rRNA gene sequence. The genomic DNA of endophytic bacteria was extracted, and 16S rDNA was amplified in Polymerase Chain Reaction (PCR) using the genomic DNA as a template and bacterial universal primers. The sequence was analyzed using a bioinformatics tool, and a phylogenetic tree was constructed using Mega X and bootstrap algorithm 17.
RESULTS AND DISCUSSION:
Preliminary Qualitative Analysis of Bioactive Compounds in the Endophytic Bacterial Extracts: From the present study, it can be inferred that all the endophytic bacterial extract showed the presence of carbohydrates and phenols in Table 1. Tannins and flavonoids were found to be present in CqB9, Cq11, and CqB12. Several secondary metabolites are alkaloids, steroids, terpenoids, peptides, polyketones, flavonoids, quinols, and phenols. These compounds also have an important role in therapeutic applications such as anticancer, antioxidant, antimicrobial, anti-inflammatory, and immunosuppressive agents 18. The presence of these metabolites indicates that the extracts of these endophytic bacteria may possess bioactivities.
TABLE 1: PRELIMINARY QUALITATIVE ANALYSIS OF BIOACTIVE COMPOUNDS IN THE ENDOPHYTIC BACTERIAL EXTRACTS
Tests | CqB1 | CqB2 | CqB3 | CqB4 | CqB5 | CqB6 | CqB7 | CqB8 | CqB9 | CqB10 | CqB11 | CqB12 | CqB13 |
Carbohydrates | + | + | + | + | + | + | + | + | + | + | + | + | + |
Tannins | - | - | - | - | - | - | - | - | + | - | + | + | - |
Flavonoids | - | - | - | - | - | + | + | - | + | - | + | + | - |
Phenols | + | + | + | + | + | + | + | + | + | + | + | + | + |
Terpenoids | - | - | - | + | + | + | - | - | + | - | + | - | - |
Steroids | - | - | - | - | - | - | - | - | + | - | - | - | - |
+ indicates presence, - indicates the absence
Determination of Total Phenolic Content: Total phenolic content of extracellular ethyl acetate extracts of 13 endophytic bacteria, estimated using the Folin-Ciocalteu’s colorimetric method, is shown in Fig. 1. Among the 13 extracellular extracts of the isolates, the highest phenolic content of 7.05 ± 0.13 mg/g was estimated in the extract of CqB9. Phenols are secondary plant metabolites that are ubiquitously present in plants and are also produced by plant-associated bacteria. The presence of such metabolites enhance the level of antioxidant 19, 20, and antagonistic activities 21, 22 of endophytic bacteria, making them effective strains for the isolation and characterization of bioactive compounds.
FIG. 1: DETERMINATION OF TOTAL PHENOLIC CONTENT
Phenolic compounds are a class of antioxidant agents that act as free radical terminators, and their bioactivities may be related to their abilities to chelate metals, inhibit lipoxygenase and scavenge free radicals 23. Antibacterial activity of phenolic compounds may be attributed to different mode of action like inhibition of synthesis of nucleic acid, destabilization, and permeabilization of cytoplasmic membrane 24.
Antimicrobial Activity: The extracellular ethyl acetate crude extracts of all the isolates were screened for antibacterial activities against Staphylococcus aureus, Bacillus subtilis, E. coli, and Klesbsiella pneumoniae. The results Table 2 revealed that the extract CqB9 possesses considerable antibacterial activity. These activities may be due to the presence of bioactive compounds. Antibacterial activities of ethyl acetate extracts of Cissus quadrangularis against Staphylococcus aureus, Escherichia coli and Bacillus subtilis has been reported 25.
TABLE 2: SCREENING FOR ANTIBACTERIAL ACTIVITIES OF EXTRACELLULAR CRUDE EXTRACTS
Test
Bacteria |
Cq
B1 |
Cq
B2 |
Cq
B3 |
Cq
B4 |
Cq
B5 |
Cq
B6 |
Cq
B7 |
Cq
B8 |
Cq
B9 |
Cq
B10 |
Cq
B11 |
Cq
B12 |
Cq
B13 |
Control |
1 | ++ | +++ | - | - | - | - | +++ | + | +++ | ++ | ++ | - | +++ | ++++ |
2 | + | ++ | - | - | - | +++ | ++ | - | +++ | - | - | - | ++ | ++++ |
3 | +++ | ++ | ++ | ++ | +++ | ++ | + | - | + | ++ | - | + | + | ++++ |
4 | +++ | - | - | ++ | ++ | ++ | - | - | +++ | - | ++ | ++ | - | ++++ |
+: Diameter of zone of inhibition </=10mm, ++: Zone of inhibition >10mm< /=15mm, +++: Zone of inhibition > 15mm, ++++: >20mm, -: no activity. 1. Bacillus subtilis, 2. Staphylococcus aureus, 3. Escherichia coli, 4. Pseudomonas aeruginosa
GC-MS: The utilization of GC-MS for the screening of different compounds is common today 26. The efficiency of separation and identification of compounds from complex biological mixtures is very high by GC-MS 27. The GC-MS of extracellular crude extract of CqB9 showed presence of 10 major compounds as indicated by some highest peaks Fig. 2. The predicted compounds and their biological activities are discussed in Table 3. GC-MS analysis indicated that the bacterium CqB9 produced bioactive compounds having antioxidant, antimicrobial, and anticancerous activities.
FIG. 2: GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS OF EXTRACELLULAR EXTRACT OF CQB9
TABLE 3: GC-MS ANALYSIS OF EXTRACELLULAR EXTRACTS OF CQB9
S. no. | Compound | Retention time in minutes | Molecular weight g/mol | Molecular Formula | Nature of the compound | Activity found |
1 | Diethyl Pthalate | 13.83 | 222 | C12H14O4
|
Aromatic diester | Antimicrobial, Antioxidant, Anticancer 28 |
2 | n-Hexadecanoic acid | 17.4 | 256 | C16H32O2 | Saturated fatty acid | Antioxidant, Pesticide, Hypocholesterolemic, Nematicide, Anti-androgenic Flavor,
Hemolytic, 5-Alpha Reductase Inhibitor, cytotoxic 28, 29 |
3 | Oleic acid | 18.3 | 282 | C18H34O2 | Unsaturated fatty acid | Antihypertensive, Increase HDL and Decrease LDL Cholesterol, Antibacterial 28, 30 |
4 | 9-Octadecenoic acid, [E]- | 19.05 | 282
|
C18H34O2
|
Elaidic acid/ trans-Oleic acid | Antidiarrhoeal 31-33 |
5 | 2,6-Bis(1,1dimethylethyl)-4phenylmethylene
cyclohexa-2,5-dien-1-one |
20.78 | 294 | C21 H26 O | Ketoenol | Not found |
6 | 1,2-Benzenedicarboxylic acid, mono(2-ethylhexyl) ester | 22.1 | 279 | C22H34O4 | Plasticizer compound | Antimicrobial 34
|
7 | 3,4-Dihydroxy-1,6-bis(3-methoxy-phenyl)-hexa-2,4-diene-1,6-dione | 31.88 | 354 | C18H12O6 | ketoenol | Not found |
8 | Isopropyl stearate | 29.45 | 326 | C21H42O2 | Stearic acid | An emollient, skin conditioning agent, binder and humectant activities 35 |
9 | But-2-endiamide, N, N’-bis(4-methoxyphenyl)- | 23.65 | 326 | C16H12N2O4 | amide | Not found |
DPPH Free Radical Scavenging Activity: DPPH radical is widely used to estimate antioxidant activity due to its ability to tap hydrogen atoms 37. The extracellular ethyl acetate extract of CqB9 exhibited significant antioxidant activity in a dose-dependent manner, as depicted in Fig. 3. Fifty percent inhibition (IC50) was achieved when the concentration of CqB9 extract was 157.5 µg. The IC50 value of 334.9 µg and 427.1 µg was reported for the endophytic bacteria Bacillus tequilensis and Bacillus subtilis, respectively isolated from a medicinal plant Fagonia indica 38. IC50 value is a widely used parameter for the measurement of free radical scavenging activity. Low IC50 indicates sig-nificant activities as compared to high IC50 value 39.
FIG. 3: DPPH FREE RADICAL SCAVENGING ASSAY OF EXTRACELLULAR EXTRACT OF ENDOPHYTE CQB9
Cytotoxic Activity: MTT assay is a well-established in-vitro method for assessing cytotoxicity against cancer cell lines. Extracts of Cissus quadrangularis have shown to inhibit the proliferation of MCF-7 cell lines 40. In order to understand the effect of extracellular ethyl acetate extract of the endophytic bacterium CqB9 isolated from Cissus quadrangularis on human breast cancer cells, experiments were conducted using cultured MCF-7 cell lines. The cytotoxic potential of the extract on MCF-7 cell lines are given in Fig. 4. MTT assay results revealed that as the concentration of CqB9 extract was increased, the percent inhibition of MCF-7 cell lines also increased.
FIG. 4: CYTOTOXIC ACTIVITY OF EXTRACELLULAR EXTRACT OF CQB9 AGAINST MCF-7 CELL LINE
The extract showed a percent inhibition of 28.33 ± 0.6 at 100 µg/ml concentration, and the percent inhibition increased to 51.42 ± 0.63 at a concentration of 1200 µg/ml. Results of cytotoxic assays showed that the extracellular ethyl acetate extract of endophytic bacterium CqB9 inhibited MCF-7 cancer cell proliferation.
16S rRNA Sequencing: The endophytic bacterium CqB9 was characterized by 16S rRNA sequencing, and a phylogenetic tree was constructed using MEGA X software and bootstrap algorithm Fig. 5. CqB9 was found to be similar to Achromobacter anxifer.
To the best of our knowledge, this is the first paper to report the isolation of endophytic bacterium from Cissus quadrangularis identified to be similar to Achromobacter anxifer.
FIG. 5: PHYLOGENETIC TREE OF THE ENDOPHYTE CQB9 BASED ON 16S rRNA SEQUENCING CONSTRUCTED USING MEGA X AND BOOTSTRAP ALGORITHM
CONCLUSION: Endophytes are considered to be potent sources of bioactive compounds. In this study, endophytic bacteria were isolated from a medicinal plant Cissus quadrangularis, and one of them was identified to be similar to Achromobacter anxifer by 16S rRNA analysis. The study shows the antibacterial and antioxidant potential of the endophytic bacteria isolated from Cissus quadrangularis. The bioactive compounds were predicted by using GCMS. The results of this study represent that endophytic bacteria may serve as a potential source of natural antibacterial, antioxidant and anticancerous compounds. The cytotoxic activity using MTT assay showed that the crude extract inhibited the proliferation of MCF-7 cell line in a dose dependent manner. However, this needs to be further confirmed by staining methods, supporting parameters like DNA fragmentation experiments and comparison with cytotoxic activity on normal cell lines. Further, the bioactive components having antibacterial, antioxidant and cytotoxic property need to be purified and studied. This study is focused on exploring endophytic bacteria from the stem of Cissus quadrangularis for their novel bioactive compounds.
In conclusion, the identification of the metabolites from the extracellular extracts of CqB9 identified as Achromobacter anxifer, predicts that this endophyte is capable of producing bioactive compounds. Further work is to be carried out to confirm the bioactivities of these metabolites. This study will provide an introduction to more comprehensive work on bioactive compounds produced by the bacterial endophytes from this important medicinal plant.
ACKNOWLEDGEMENT: The authors thank former and present Head, the Faculty members, supporting staff of the Department of Plant Biology and Plant Biotechnology and the Principal, Ethiraj College for Women, (autonomous) Chennai - 600 008, for their valuable support and encouragement throughout the entire period of research. We would also like to express our thanks for the facilities extended by the Central Instrumentation Centre of Ethiraj College for Women. Sincere thanks to IIT Madras for the extension of facilities.
CONFLICTS OF INTEREST: Authors declare no conflict of interest.
REFERENCES:
- Fazilath U, Mohan CD, Chandra NS and Srinivas C: Advances in Endophytic Fungal Research. Springer, Edition 1, 2019: 243-65.
- Alwin A, Kristin IM and Bret AN: Exploring the potential of endophytes from medicinal plants as a source of antimycobacterial compounds. Microbiol Res 2014; 169: 483-95.
- Siddiqua A and Mittapally S: A review on Cissus quadrangularis. The Pharma Innovation Journal 2017; 6(7): 329-34.
- Thalkari AB, Karwa PN, Sagde RM, Chopane PS and Zhambare KK: Cissus quadrangularis: A Natural Booster. Res J Pharmacognosy and Phytochem 2019; 11(3): 129-36.
- Tharshanodayan NJQ and Rohini P: A review on medicinal properties and nutraceutical importance of Cissus quadrangularis. Int J of Life Science 2019; 7(2): 368-74.
- Kuppuramalingam AP and Ramesh B: Antioxidant activity of Cissus quadrangularis stem in-vitro. World Journal of Pharmaceutical Research 2018; 7(11): 759-65.
- Sasi B and Tamizhiniyan P: Antioxidant properties of various parts Cissus quadrangularis in different solvents. Journal of Medicinal Botany 2018; 2: 27-33.
- Suresh P, Xavier AS, Karthik VP and Punnagai K: Anticancer activity of Cissus quadrangularis methanolic extract against MG63 human osteosarcoma cells – an in-vitro evaluation using cytotoxicity assay. Biomed Pharmacol J 2019; 12(2): 975-80.
- Geetha S and Vasuki R: Anticancer activity of Cissus quadrangularis. International Journal of Innovative Technology and Exploring Engineering 2019; 8(6): 603-05.
- Strobel G and Daisy B: Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 2003; 67: 491-502.
- Raaman N: Phytochemical Techniques. New India Publishing Agency, New Delhi, 2006: 19-25.
- Harborne JB: Phytochemical methods: a guide to modern techniques of plant analysis. Chapman and Hall Limited, London, Third Edition 1998.
- Slinkard K and Singleton VL: Total phenol analysis: Automation and comparison with manual methods. Am J Enol Viticult 1977; 28: 49-55.
- Perez C, Pauli M and Bazerque P: An Antibiotic Assay by Agar Well Diffusion Method. Acta Biologiae et Medicinae Experimentalis 1990; 15: 113-15.
- Blois MS: Antioxidant Determinations by the use of a stable free radical. Nature 1958; 181: 1199-1200.
- Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.
- Kumar S, Stecher G, Li M, Knyaz C and Tamura K: MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Mol Biol Evol 2018; 35: 1547-49.
- Seca AML and Pinto DCGA: Biological potential and medical use of secondary metabolites. Medicines 2019; 6(2): 66.
- Kaurinovic B and Vastag D: Antioxidants, IntechOpen, London 2019. doi 10.5772/intechopen.83731.
- Liaudanskas M, Zymone K, Viskelis J, Klevinskas A and Janulis V: Determination of the phenolic composition and antioxidant activity of pear extracts. Hindawi Journal of Chemistry 2017; 1-9.
- Aldulaimi OA: General overview of phenolics from plant to laboratory, good antibacterials or not. Pharmacognosy Reviews 2017; 11(22): 123-27.
- Pereira JA, Oliveira I, Sousa A, Valento P, Andrade PB, Ferreira IC, Ferreres F, Bento A, Seabra R and Estivinho L: Walnut (Juglans regia) leaves: phenolic compounds, antimicrobial activity and antioxidant potential of different cultivars. Food Chem Toxico 2007; 45: 2287-95.
- Ramesha A and Srinivas C: Antimicrobial activity and phytochemical analysis of crude extracts of endophytic fungi isolated from Plumeria acuminata and Plumeria obtusifolia L. Euro J Exp. Bio 2014; 4: 35-43.
- Gorniak I, Bartoszewski R and Kroliczewski J: Comprehensive review of antimicrobial activities of plant flavonoids. J. Phytochem Rev 2019; 18: 241-72.
- Selvamaleeswaran P, Manimaran E and Sureshkumar M: Antimicrobial activity of medicinally important plant-Cissus quadrangularis Linn against some pathogenic bacteria. Journal of Chemical and Pharmaceutical Research 2016; 8(11): 92-96
- Tiwari V, Roy R and Tiwari M: Antimicrobial active herbal compounds against Acinetobacter baumannii and other pathogens. Front. Microbiol 2015; 6: 618.
- Al-Rubaye AF, Hameed IH and Kadhim MJ: A review: uses of gas chromatography-mass spectrometry (GC-MS) technique for analysis of bioactive natural compounds of some plants. International Journal of Toxicological and Pharmacological Research 2017; 9(1): 81-85.
- Singaravadivel C and Santhanaraj KJ: Gas chromato-graphy and mass spectroscopic determination of phyto-compounds in Cissus vitiginea leaf: Der Pharmacia Lettre 2016; 8: 292-97.
- Tulika T and Agarwal M: Phytochemical and GC-MS analysis of bioactive constituents in the ethanolic of Pistia stratiotes and Eichhornia crassipes (Mart.) solms. J of Pharmacognosy and Phytochemistry 2017; 6: 195-206.
- Yoon BK., Jackman JA, Valle-Gonzalez ER and Cho NJ: Antibacterial free fatty acids and monoglycerides: biological activities, experimental testing, and therapeutic applications. International Journal of Molecular Sciences 2018; 19(4): 1114.
- Ramachandran P and Elezabeth DVS: GC–MS study on ethanolic extract of Vetiveria lawsonii The Journal of Phytopharmacology 2016; 5(3): 108-11.
- Bentrad N, Terrak G R, Benmalek Y and Rahmania F: Studies on chemical composition and antimicrobial activities of bioactive molecules from date palm (Phoenix dactylifera) Pollens and seeds. Afr J Tradit Complement Altern Med 2017; 14: 242-26.
- Garba S and Garba I: Anti-Diarrhoeal Properties of Cis-9-octadecenoic acid isolated from Landolphia owariensis Organic& Medicinal Chem IJ 2017; 3: 555619.
- Adeyemi MA, Ekunseitan DA, Abiola SS, Dipeolu MA, Egbeyale LT and Sogunle OM: Phytochemical analysis and GC-MS determination of Lagenaria breviflora fruit. International Journal of Pharmacognosy and Phytochemical Research 2017; 9(7): 1045-50.
- Elaiyaraja A and Chandramohan G: Comparative phytochemical profile of Indoneesiella echioides (L.) Nees leaves using GC-MS. J Pharmacogn and Phytochem 2016; 5: 158-71.
- Ariharan VN, Kalirajan K and Prasad NP: GC-MS determination of bioactive components of three traits of vilvam (bael). Asian J Pharm Clin Res 2015; 8: 338-42.
- Gupta D: Methods for determination of antioxidant capacity: a review. Int J Pharm Sci Res 2015; 6(2): 546-66.
- Rahman L, Shinwari KZ, Iqrar I, Lutfur R and Tanveer F. An assessment on the role of endophytic microbes in the therapeutic potential of Fagonia indica. Ann Clin Microbiol Antimicrob 2017; 16: 53.
- Fidrianny I, Budiana W and Ruslan K: Antioxidant activities of various extracts from Ardisia sp leaves using dpph and cuprac assays and correlation with total flavonoid, phenolic, carotenoid content. International Journal of Pharmacognosy and Phytochemical Research 2015; 7(4): 859-65.
- Subramani B: Analysis of potential toxicological, phytochemical and anticancer properties from Cissus quandrangularis. Research Journal of Lifesciences, Bioinformatics, Pharmaceutical and Chemical Sciences 2018; 4(3): 94-105.
How to cite this article:
Anwar PZ, Sezhian UG and Narasingam A: Bioactive compound analysis and bioactivities of endophytic bacteria from Cissus quadrangularis. Int J Pharm Sci & Res 2020; 11(11): 5553-60. doi: 10.13040/IJPSR.0975-8232.11(11).5553-60.
All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
Article Information
25
5553-5560
749
737
English
IJPSR
P. Z. Anwar, U. G. Sezhian * and A. Narasingam
Department of Plant Biology and Plant Biotechnology, Ethiraj College for Women (Autonomous), Chennai, Tamil Nadu, India.
umasezhian@gmail.com
11 November 2019
20 February 2020
21 March 2020
10.13040/IJPSR.0975-8232.11(11).5553-60
01 November 2020