192
R. Baishya and S. Banerjee
2.6 Quantification of EPS Components
EPS was quantified by measuring proteins by Lowry’s method and carbohydrates by
Anthrone method (Teanpaisan et al. 2016).
2.7 Emulsifying Activity
The EPS solutions were heated and then cooled and volume was made up to 2 ml
using then phosphate-buffered saline (PBS). Olive oil was added and vortexed for
1 min and absorbance was measured at 540 nm after 30 min (Kavita et al. 2014).
2.8 Protein Profile Analysis by SDS-PAGE
EPS proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoresis were performed in 5% stacking gel and
10% separating gel in Tris-Glycine-SDS buffer and was run with a constant voltage of
20 V/cm. The gels were then stained with Coomassie Brilliant Blue R-250 (Sambrook
et al. 1989). The molecular weight of the proteins bands was compared with the
standard protein ladder (10–245 kDa) to know its respective sizes.
3 Results and Discussion
3.1 Antimicrobial Activity
The antimicrobial potential of all the five plant extracts against 30 strong biofilmforming clinical strains was evaluated according to their zone of inhibition. All the 30
strains were susceptible to Curcuma longa (haldi), Azadirachta indica (neem), 71%
samples were found susceptible to Ocimum sanctum (tulsi) and Zingiber officinale
(ginger), and 93.33% were susceptible to Allium sativum (Garlic) (Fig. 1).
3.2 Study of Anti-biofilm Activity of Plant Extract
The anti-biofilm activity of plant extracts on the selected strain S. aureus QS1 by
crystal violet method is shown in Fig. 2. For tulsi, there is a gradual increase of
anti-adherence activity from 10 to 38%; for neem, the anti-adherence activity was
R. Baishya and S. Banerjee
2.6 Quantification of EPS Components
EPS was quantified by measuring proteins by Lowry’s method and carbohydrates by
Anthrone method (Teanpaisan et al. 2016).
2.7 Emulsifying Activity
The EPS solutions were heated and then cooled and volume was made up to 2 ml
using then phosphate-buffered saline (PBS). Olive oil was added and vortexed for
1 min and absorbance was measured at 540 nm after 30 min (Kavita et al. 2014).
2.8 Protein Profile Analysis by SDS-PAGE
EPS proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoresis were performed in 5% stacking gel and
10% separating gel in Tris-Glycine-SDS buffer and was run with a constant voltage of
20 V/cm. The gels were then stained with Coomassie Brilliant Blue R-250 (Sambrook
et al. 1989). The molecular weight of the proteins bands was compared with the
standard protein ladder (10–245 kDa) to know its respective sizes.
3 Results and Discussion
3.1 Antimicrobial Activity
The antimicrobial potential of all the five plant extracts against 30 strong biofilmforming clinical strains was evaluated according to their zone of inhibition. All the 30
strains were susceptible to Curcuma longa (haldi), Azadirachta indica (neem), 71%
samples were found susceptible to Ocimum sanctum (tulsi) and Zingiber officinale
(ginger), and 93.33% were susceptible to Allium sativum (Garlic) (Fig. 1).
3.2 Study of Anti-biofilm Activity of Plant Extract
The anti-biofilm activity of plant extracts on the selected strain S. aureus QS1 by
crystal violet method is shown in Fig. 2. For tulsi, there is a gradual increase of
anti-adherence activity from 10 to 38%; for neem, the anti-adherence activity was