Antimicrobial activity of kojic acid from endophytic fungus Colletotrichum gloeosporioides isolated from Sonneratia apetala,a mangrove plant of the Sundarbans
|
摘要
Objective: To isolate and evaluate the antimicrobial activity of the active principle(s) from the ethyl acetate(EtOAc) extract of endophytic fungus Colietotrichum gloeosporioides(C.gloeosporioides) isolated from Sonneratia apetala. Methods: Water agar technique was used to isolate the fungus, and both microscopic and molecular techniques were used for identification of the strain. Potato dextrose broth was used to grow the fungus in large-scale. Reversed-phase preparative HPLC analysis was performed to isolate the major active compound, kojic acid. The EtOAc extract and kojic acid were screened for their antimicrobial activity against two Grampositive and two Gram-negative bacteria as well as a fungal strain using the resazurin 96-well microtitre plate antimicrobial assay. Results: The fungus C. gloeosporioides was isolated from the leaves of Sonneratia apetala. Initial identification of the fugal isolate was carried out using spore characteristics observed under the microscope. Subsequently, the ITS1-5.8 S-ITS2 sequencing was employed for species-level identification of the fungus C. gloeosporioides. Five litres of liquid culture of the fungus produced approximately 610 mg of a mixture of secondary metabolites.Kojic acid(1) was isolated as the main secondary metabolite present in the fungal extract, and the structure was confirmed by 1 D, 2 D NMR and mass spectrometry. The EtOAc extract and compound 1 exhibited considerable antimicrobial activity against all tested microorganisms.Whilst the minimum inhibitory concentration(MIC) values from the EtOAc extract ranged between 2.4×10~(-4)mg/mL and 2.5 mg/mL, those of kojic acid(1) were between 0.125 mg/mL and1 mg/mL. The EtOAc extract and kojic acid(1) were most active against Pseudomonas aeruginosa(MIC = 2.4×10~(-4). mg/mL) and Micrococcus luteus(MIC = 0.125 mg/mL), respectively. Conclusions:The results revealed that the endophytic fungus C. gloeosporioides could be a good source of commercially important kojic acid, which exhibited antimicrobial properties.
Objective: To isolate and evaluate the antimicrobial activity of the active principle(s) from the ethyl acetate(EtOAc) extract of endophytic fungus Colietotrichum gloeosporioides(C.gloeosporioides) isolated from Sonneratia apetala. Methods: Water agar technique was used to isolate the fungus, and both microscopic and molecular techniques were used for identification of the strain. Potato dextrose broth was used to grow the fungus in large-scale. Reversed-phase preparative HPLC analysis was performed to isolate the major active compound, kojic acid. The EtOAc extract and kojic acid were screened for their antimicrobial activity against two Grampositive and two Gram-negative bacteria as well as a fungal strain using the resazurin 96-well microtitre plate antimicrobial assay. Results: The fungus C. gloeosporioides was isolated from the leaves of Sonneratia apetala. Initial identification of the fugal isolate was carried out using spore characteristics observed under the microscope. Subsequently, the ITS1-5.8 S-ITS2 sequencing was employed for species-level identification of the fungus C. gloeosporioides. Five litres of liquid culture of the fungus produced approximately 610 mg of a mixture of secondary metabolites.Kojic acid(1) was isolated as the main secondary metabolite present in the fungal extract, and the structure was confirmed by 1 D, 2 D NMR and mass spectrometry. The EtOAc extract and compound 1 exhibited considerable antimicrobial activity against all tested microorganisms.Whilst the minimum inhibitory concentration(MIC) values from the EtOAc extract ranged between 2.4×10~(-4)mg/mL and 2.5 mg/mL, those of kojic acid(1) were between 0.125 mg/mL and1 mg/mL. The EtOAc extract and kojic acid(1) were most active against Pseudomonas aeruginosa(MIC = 2.4×10~(-4). mg/mL) and Micrococcus luteus(MIC = 0.125 mg/mL), respectively. Conclusions:The results revealed that the endophytic fungus C. gloeosporioides could be a good source of commercially important kojic acid, which exhibited antimicrobial properties.
Objective: To isolate and evaluate the antimicrobial activity of the active principle(s) from the ethyl acetate(EtOAc) extract of endophytic fungus Colietotrichum gloeosporioides(C.gloeosporioides) isolated from Sonneratia apetala. Methods: Water agar technique was used to isolate the fungus, and both microscopic and molecular techniques were used for identification of the strain. Potato dextrose broth was used to grow the fungus in large-scale. Reversed-phase preparative HPLC analysis was performed to isolate the major active compound, kojic acid. The EtOAc extract and kojic acid were screened for their antimicrobial activity against two Grampositive and two Gram-negative bacteria as well as a fungal strain using the resazurin 96-well microtitre plate antimicrobial assay. Results: The fungus C. gloeosporioides was isolated from the leaves of Sonneratia apetala. Initial identification of the fugal isolate was carried out using spore characteristics observed under the microscope. Subsequently, the ITS1-5.8 S-ITS2 sequencing was employed for species-level identification of the fungus C. gloeosporioides. Five litres of liquid culture of the fungus produced approximately 610 mg of a mixture of secondary metabolites.Kojic acid(1) was isolated as the main secondary metabolite present in the fungal extract, and the structure was confirmed by 1 D, 2 D NMR and mass spectrometry. The EtOAc extract and compound 1 exhibited considerable antimicrobial activity against all tested microorganisms.Whilst the minimum inhibitory concentration(MIC) values from the EtOAc extract ranged between 2.4×10~(-4)mg/mL and 2.5 mg/mL, those of kojic acid(1) were between 0.125 mg/mL and1 mg/mL. The EtOAc extract and kojic acid(1) were most active against Pseudomonas aeruginosa(MIC = 2.4×10~(-4). mg/mL) and Micrococcus luteus(MIC = 0.125 mg/mL), respectively. Conclusions:The results revealed that the endophytic fungus C. gloeosporioides could be a good source of commercially important kojic acid, which exhibited antimicrobial properties.
引文
[1]Premanathan M,Arakaki R,Izumi H,Kathiresan K,Nakano M,Yamamoto N,et al.Antiviral properties of a mangrove plant,Rhizophora apiculata against immunodeficiency virus.Antiviral Res 1999;44:113-122.
[2]Bandaranayake WM.Bioactivities,bioactive compounds and chemical constituents of mangrove plants.Wetlands Ecol Management 2002;10:421-452.
[3]Jamale B,Joshi GV.Effect on age of mineral constituents.Polyphenoloxides and peroxides in mangrove leaves.Indian J Exp Biol1998;16:117-120.
[4]Strobel GA,Yang X,Sears J,Kramer R,Sidhu R,Hess WM.Taxol from Pestalotiopsis microspora,an endophytic fungus from Taxus wallacbiana.Microbiol 1996;142:435-440.
[5]Bhimba BV,Agnel Defora Franco DA,Mathew JM,Jose GM,Joel EL,Thangaraj M.Anticancer and antimicrobial activity of mangrove derived fungi Hypocrea lixii.Chin J Nat Med 2012;10:77-80.
[6]Nurunnabi TR,Nahar L,Al-Majmaie S,Rahman SMM,Sohrab MH,Billah MM,et al.Anti-MRSA activity of oxysporone and xylitol from the endophytic fungus Pestalotia sp.growing on the Sundarbans mangrove plant Heritiera fomes.Phytother Res 2018;32(2):348-354.
[7]Ren H,Lu H,Shen W,Huang C,Guo Q,Li Z,et al.Sonneratia apetala Buch.Ham in the mangrove ecosystem of China:An invasive species or restoration species?Ecol Eng 2009;35:1243-1248.
[8]Bandaranayake WM.Survey of mangrove plants from northern Australia for phytochemical constituents and UV-absorbing compounds.Curr TopPhytochem 1995;14:69-78.
[9]Hossain SJ,Islam MR,Pervin T,Iftekharuzzaman M,Hamdi OAA,Mubassara S,et al.Antibacterial,anti-diarrhoeal,analgesic,cytotoxic activities and GC-MS profiling of Sonneratia apetala(Buch.-Ham.)seed.Prev Nutr Food Sci 2017;22:157-165.
[10]Zabala NQ.Silviculture of species.Development of professional education in the Forestry Sector of Bangladesh.Rome:Food and Agriculture Organization of the United Nations;1990.
[11]Qadri M,Johri S,Shah BA,Khajuria A,Sidiq T,Lattoo SK,et al.Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas.Springerplus 2013;2:1-14.
[12]Sarker SD,Nahar L,Kumarasamy Y.Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth,and its application in the in vitro antibacterial screening of phytochemicals.Methods 2007;42:321-324.
[13]Eyong KO,Ambassa P,Yimdjo MC,Sidjui LS,Folefoc GN.A new source of kojic acid isolated from Kigelia africana:A possible precursor for quinone biosynthesis.RASAYAN J Chem 2012;5:477-480.
[14]Cai L,Hyde KD,Taylor PWJ,Weir BS,Waller J,Abang MM,et al.A polyphasic approach for studying Colletotrichum.Fungal Divers 2009:39:183-204.
[15]Crouch JA,Clarke BB,White JW,Hillman BI.Systematic analysis of the falcate-spored graminicolous Colletotrichum and a description of six new species of the fungus from warm-season grasses.Mycologia 2009;101:717-732.
[16]Damm U,Woudenberg JHC,Cannon PF,Crous PW.Colletotrichum species with curved conidia from herbaceous hosts.Fungal Divers 2009;39:45-87.
[17]Damm U,Baroncelli R,Cai L,Kubo Y,O-Connell R,Weir B,et al.Colletotrichum:Species,ecology and interactions.IMA Fungus 2010;1:161-165.
[18]Zhang JJ,Chen H,Chen MJ,Wang H,Wang Q,Song XX,et al.Kojic acid mediated damage responses induce mycelial regeneration in the basidiomycete Hypsizygus marmoreus.PLoS One 2017;12(11):e0187351.
[19]Wang Y,Xiu L,Gao W,Niu L,Huang C,Yang P.et al.Isoprenylated phenolic compounds from Morus macroura as potent tyrosinase inhibitor.Planta Med 2018;84(5):336-343.
[20]Gust PJ,Luke JD.Kojic acid.J Dermatol Nurses Assoc 2016;8:338-340.
[21]Sarkar R,Gokhale N,Godse K,Ailawadi P,Aya L,Sarma N,et al.Medical management of melisma:A review with consensus recommendation by Indian pigmentary expert group.Indian J Dermatol2017;62:450-469.
[22]Kim TY,Jang JY,Jeon SJ,Lee HW,Bae CH,Yeo JH,et al.Nematicidal activity of kojic acid produced by Aspergillus oryzae against Meloidogyne incognita.J Microbiol Biotechnol 2016;26:1383-1391.
[23]Wei SP,Ji ZQ.Isolation,synthesis and antifungal activity of kojic acid and its derivative.Chem Nat Compd 2016;52:123-127.
[24]Rodrigues APD,Farias LHS,Carvalho ASC,Santos AS,do Nascimento JLM,Silva EO.A novel function for kojic acid,a secondary metabolite from Aspergillus fungi,as antileishmanial agent.PLoS One 2014;9:e9125.
[2]Bandaranayake WM.Bioactivities,bioactive compounds and chemical constituents of mangrove plants.Wetlands Ecol Management 2002;10:421-452.
[3]Jamale B,Joshi GV.Effect on age of mineral constituents.Polyphenoloxides and peroxides in mangrove leaves.Indian J Exp Biol1998;16:117-120.
[4]Strobel GA,Yang X,Sears J,Kramer R,Sidhu R,Hess WM.Taxol from Pestalotiopsis microspora,an endophytic fungus from Taxus wallacbiana.Microbiol 1996;142:435-440.
[5]Bhimba BV,Agnel Defora Franco DA,Mathew JM,Jose GM,Joel EL,Thangaraj M.Anticancer and antimicrobial activity of mangrove derived fungi Hypocrea lixii.Chin J Nat Med 2012;10:77-80.
[6]Nurunnabi TR,Nahar L,Al-Majmaie S,Rahman SMM,Sohrab MH,Billah MM,et al.Anti-MRSA activity of oxysporone and xylitol from the endophytic fungus Pestalotia sp.growing on the Sundarbans mangrove plant Heritiera fomes.Phytother Res 2018;32(2):348-354.
[7]Ren H,Lu H,Shen W,Huang C,Guo Q,Li Z,et al.Sonneratia apetala Buch.Ham in the mangrove ecosystem of China:An invasive species or restoration species?Ecol Eng 2009;35:1243-1248.
[8]Bandaranayake WM.Survey of mangrove plants from northern Australia for phytochemical constituents and UV-absorbing compounds.Curr TopPhytochem 1995;14:69-78.
[9]Hossain SJ,Islam MR,Pervin T,Iftekharuzzaman M,Hamdi OAA,Mubassara S,et al.Antibacterial,anti-diarrhoeal,analgesic,cytotoxic activities and GC-MS profiling of Sonneratia apetala(Buch.-Ham.)seed.Prev Nutr Food Sci 2017;22:157-165.
[10]Zabala NQ.Silviculture of species.Development of professional education in the Forestry Sector of Bangladesh.Rome:Food and Agriculture Organization of the United Nations;1990.
[11]Qadri M,Johri S,Shah BA,Khajuria A,Sidiq T,Lattoo SK,et al.Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas.Springerplus 2013;2:1-14.
[12]Sarker SD,Nahar L,Kumarasamy Y.Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth,and its application in the in vitro antibacterial screening of phytochemicals.Methods 2007;42:321-324.
[13]Eyong KO,Ambassa P,Yimdjo MC,Sidjui LS,Folefoc GN.A new source of kojic acid isolated from Kigelia africana:A possible precursor for quinone biosynthesis.RASAYAN J Chem 2012;5:477-480.
[14]Cai L,Hyde KD,Taylor PWJ,Weir BS,Waller J,Abang MM,et al.A polyphasic approach for studying Colletotrichum.Fungal Divers 2009:39:183-204.
[15]Crouch JA,Clarke BB,White JW,Hillman BI.Systematic analysis of the falcate-spored graminicolous Colletotrichum and a description of six new species of the fungus from warm-season grasses.Mycologia 2009;101:717-732.
[16]Damm U,Woudenberg JHC,Cannon PF,Crous PW.Colletotrichum species with curved conidia from herbaceous hosts.Fungal Divers 2009;39:45-87.
[17]Damm U,Baroncelli R,Cai L,Kubo Y,O-Connell R,Weir B,et al.Colletotrichum:Species,ecology and interactions.IMA Fungus 2010;1:161-165.
[18]Zhang JJ,Chen H,Chen MJ,Wang H,Wang Q,Song XX,et al.Kojic acid mediated damage responses induce mycelial regeneration in the basidiomycete Hypsizygus marmoreus.PLoS One 2017;12(11):e0187351.
[19]Wang Y,Xiu L,Gao W,Niu L,Huang C,Yang P.et al.Isoprenylated phenolic compounds from Morus macroura as potent tyrosinase inhibitor.Planta Med 2018;84(5):336-343.
[20]Gust PJ,Luke JD.Kojic acid.J Dermatol Nurses Assoc 2016;8:338-340.
[21]Sarkar R,Gokhale N,Godse K,Ailawadi P,Aya L,Sarma N,et al.Medical management of melisma:A review with consensus recommendation by Indian pigmentary expert group.Indian J Dermatol2017;62:450-469.
[22]Kim TY,Jang JY,Jeon SJ,Lee HW,Bae CH,Yeo JH,et al.Nematicidal activity of kojic acid produced by Aspergillus oryzae against Meloidogyne incognita.J Microbiol Biotechnol 2016;26:1383-1391.
[23]Wei SP,Ji ZQ.Isolation,synthesis and antifungal activity of kojic acid and its derivative.Chem Nat Compd 2016;52:123-127.
[24]Rodrigues APD,Farias LHS,Carvalho ASC,Santos AS,do Nascimento JLM,Silva EO.A novel function for kojic acid,a secondary metabolite from Aspergillus fungi,as antileishmanial agent.PLoS One 2014;9:e9125.