Limits...
Molecular characterization of an endophytic Phomopsis liquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites

View Article: PubMed Central - PubMed

ABSTRACT

An endophytic fungus Phomopsis liquidambaris CBR-15, was isolated from Cryptolepis buchanani Roem. (Asclepiadaceae) and identified by its characteristic culture morphology and molecular analysis of the ITS region of rDNA and intervening 5.8S rRNA gene. The impact of different culture media on biosynthesis of antimicrobial metabolites was tested by disc diffusion assay. Polyketide synthase gene (PKS) of the endophytic fungus was investigated using three pairs of degenerate primers LC1–LC2c, LC3–LC5c and KS3–KS4c by PCR. TLC-bioautography method was employed to detect the antimicrobial metabolites. Antimicrobial metabolites fractionated with ethyl acetate extract showed significant antimicrobial activity against the test bacteria and fungi. Biosynthesis of antimicrobial metabolites was optimum as depicted by zone of inhibition from ethyl acetate extract cultured in potato dextrose broth. Strain CBR-15 was identified as Phomopsisliquidambaris and PKS genes of the fungus were amplified with LC3–LC5c and KS3–KS4c sets of degenerate primers. These findings suggest that endophytic P.liquidambaris CBR-15 harbor iterative type I fungal PKS gene domain which indicates the biosynthetic potential of endophytic fungi as producers of natural antimicrobial metabolites. The study also demonstrates the utilization and optimization of different culture media which best supports for the biosynthesis of the antimicrobial metabolites from P.liquidambaris.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-014-0204-2) contains supplementary material, which is available to authorized users.

No MeSH data available.


Antimicrobial activity of ethyl acetate extract of P.liquidambaris CBR-15 cultured in different media by disc diffusion assay against E. coli (2a and 2b) and B. subtilis (2c and 2d) where+VE  positive control,−VE  negative control and PDB,MPY, YSB, MCB are the different culture media (see “Materials and methods” for abbreviation) extract of Phomopsisliquidambari CBR-15
© Copyright Policy - OpenAccess
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4362740&req=5

Fig2: Antimicrobial activity of ethyl acetate extract of P.liquidambaris CBR-15 cultured in different media by disc diffusion assay against E. coli (2a and 2b) and B. subtilis (2c and 2d) where+VE  positive control,−VE  negative control and PDB,MPY, YSB, MCB are the different culture media (see “Materials and methods” for abbreviation) extract of Phomopsisliquidambari CBR-15

Mentions: During disc diffusion assay, antimicrobial activity of ethyl acetate extracts cultured in different media were analyzed. The utilization of different mycological media as nutritional supplements can impact on the production of bioactive secondary metabolites. Application of multiple fermentation conditions is the desirable method that could enhance the probability of successful discovery of bioactive metabolites from a given strain (Bills et al. 2008). One such way to trigger the production of secondary metabolites is to vary the medium composition. The principle behind this method, named as one strain—many compounds (OSMAC) approach, is to expose the microorganism to other cultivating conditions than the standards used in laboratories (Fuchser and Zeeck 1997; Schiewe and Zeeck 1999; Hofs et al. 2000; Bills et al. 2008). Media composition, temperature, pH, culture vessel, aeration, cultivation time, light intensity can increase or reduce the production of the bioactive compounds by the strain (Bode et al. 2002; Siqueria et al. 2011). Yenn et al. (2012) reported anti-candidal activity of Phomopsis sp. ED2 cultured in yeast extract sucrose (YES) broth with aqueous extract of host plant. However, understanding of the exact mechanisms for the change in metabolic profile due to change in culture or fermentation conditions is usually not completely understood and therefore difficult to predict (Bode et al. 2002). This work demonstrates that PDB serves as optimum culture media for the biosynthesis of antimicrobial metabolites which facilitates isolation and characterization of antimicrobial metabolites from P.liquidambaris (Fig. 2).Fig. 2


Molecular characterization of an endophytic Phomopsis liquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites
Antimicrobial activity of ethyl acetate extract of P.liquidambaris CBR-15 cultured in different media by disc diffusion assay against E. coli (2a and 2b) and B. subtilis (2c and 2d) where+VE  positive control,−VE  negative control and PDB,MPY, YSB, MCB are the different culture media (see “Materials and methods” for abbreviation) extract of Phomopsisliquidambari CBR-15
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4362740&req=5

Fig2: Antimicrobial activity of ethyl acetate extract of P.liquidambaris CBR-15 cultured in different media by disc diffusion assay against E. coli (2a and 2b) and B. subtilis (2c and 2d) where+VE  positive control,−VE  negative control and PDB,MPY, YSB, MCB are the different culture media (see “Materials and methods” for abbreviation) extract of Phomopsisliquidambari CBR-15
Mentions: During disc diffusion assay, antimicrobial activity of ethyl acetate extracts cultured in different media were analyzed. The utilization of different mycological media as nutritional supplements can impact on the production of bioactive secondary metabolites. Application of multiple fermentation conditions is the desirable method that could enhance the probability of successful discovery of bioactive metabolites from a given strain (Bills et al. 2008). One such way to trigger the production of secondary metabolites is to vary the medium composition. The principle behind this method, named as one strain—many compounds (OSMAC) approach, is to expose the microorganism to other cultivating conditions than the standards used in laboratories (Fuchser and Zeeck 1997; Schiewe and Zeeck 1999; Hofs et al. 2000; Bills et al. 2008). Media composition, temperature, pH, culture vessel, aeration, cultivation time, light intensity can increase or reduce the production of the bioactive compounds by the strain (Bode et al. 2002; Siqueria et al. 2011). Yenn et al. (2012) reported anti-candidal activity of Phomopsis sp. ED2 cultured in yeast extract sucrose (YES) broth with aqueous extract of host plant. However, understanding of the exact mechanisms for the change in metabolic profile due to change in culture or fermentation conditions is usually not completely understood and therefore difficult to predict (Bode et al. 2002). This work demonstrates that PDB serves as optimum culture media for the biosynthesis of antimicrobial metabolites which facilitates isolation and characterization of antimicrobial metabolites from P.liquidambaris (Fig. 2).Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

An endophytic fungus Phomopsis liquidambaris CBR-15, was isolated from Cryptolepis buchanani Roem. (Asclepiadaceae) and identified by its characteristic culture morphology and molecular analysis of the ITS region of rDNA and intervening 5.8S rRNA gene. The impact of different culture media on biosynthesis of antimicrobial metabolites was tested by disc diffusion assay. Polyketide synthase gene (PKS) of the endophytic fungus was investigated using three pairs of degenerate primers LC1–LC2c, LC3–LC5c and KS3–KS4c by PCR. TLC-bioautography method was employed to detect the antimicrobial metabolites. Antimicrobial metabolites fractionated with ethyl acetate extract showed significant antimicrobial activity against the test bacteria and fungi. Biosynthesis of antimicrobial metabolites was optimum as depicted by zone of inhibition from ethyl acetate extract cultured in potato dextrose broth. Strain CBR-15 was identified as Phomopsisliquidambaris and PKS genes of the fungus were amplified with LC3–LC5c and KS3–KS4c sets of degenerate primers. These findings suggest that endophytic P.liquidambaris CBR-15 harbor iterative type I fungal PKS gene domain which indicates the biosynthetic potential of endophytic fungi as producers of natural antimicrobial metabolites. The study also demonstrates the utilization and optimization of different culture media which best supports for the biosynthesis of the antimicrobial metabolites from P.liquidambaris.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-014-0204-2) contains supplementary material, which is available to authorized users.

No MeSH data available.