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. 2015 Jan 21:5:753.
doi: 10.3389/fmicb.2014.00753. eCollection 2014.

Characterization of saltern based Streptomyces sp. and statistical media optimization for its improved antibacterial activity

Pandiyan Rajeswari  1 , Polpass Arul Jose  1 , Richa Amiya  1 , Solomon Robinson David Jebakumar  1
Affiliations

Characterization of saltern based Streptomyces sp. and statistical media optimization for its improved antibacterial activity

Pandiyan Rajeswari et al. Front Microbiol. .

Abstract

A moderately halotolerant Streptomyces strain, designated JAJ13 was characterized and a culture medium was statistically optimized to improve its antibacterial activity. Based on the phenotypic and molecular characteristics, strain JAJ13 was identified as a moderately halotolerant Streptomyces sp. JAJ13. Novelty of the strain JAJ13 in production of antibacterial compound was assessed by sequence analysis of KSα gene and LC-MS analysis of the active compound. Optimization of the culture medium for antibacterial compound production by the strain JAJ13 was performed with statistical methodology based on experimental designs. Initially, a starch based basal production medium was selected out of eight different production media screened for antibacterial compound production by Streptomyces sp. JAJ13. Plackett-Burman design was employed to screen the influential media components affecting the antibacterial compound production. Subsequently, statistical optimization of selected medium components was performed by employing the response surface methodology (RSM) with Box-Behnken design. The optimum initial level of CuSO4.5H2O, (NH4)2SO4 and K2HPO4 for the highest antibacterial activity was determined to be at 4.45 mg, 1.96 g, and 1.15 g in 1 L of distilled H2O, respectively. PBD and RSM guided design of experiments resulted in a maximum antibacterial activity of 23.37 ± 2.08 mm, which is a 78.8% increase in comparison with that obtained in the unoptimized medium. This study points the success of statistical model in developing an optimized production media for enhanced antibacterial compound production by Streptomyces sp. JAJ13.

Keywords: Box-Behnken design; Plackett-Burman design; Streptomyces; antibiotics; response surface methodology; solar saltern.

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Figures

Figure 1
Figure 1
Growth and morphology of strain JAJ13 on different ISP media.
Figure 2
Figure 2
Neighbor-joining phylogenetic tree based on 16S rRNA gene sequence of JAJ13 shows the relationships between JAJ13 and related species of the genus Streptomyces. Actinopolyspora mortivallisT was used as out-group. Numbers at nodes indicate the levels of bootstrap support (%) based on a neighbor-joining analysis of 1000 resampled datasets; only values above 50% are shown. Score bar represents 0.01 substitutions per site.
Figure 3
Figure 3
Mass spectrum of antibacterial compound detected in Streptomyces sp. JAJ13.
Figure 4
Figure 4
Effect of eight different production media on antibacterial compound production in Streptomyces sp. JAJ13.
Figure 5
Figure 5
Pareto chart showing the effect of different variables (media components) on antibacterial compound production.
Figure 6
Figure 6
Diagnostic plots showing the model adequacy. (A) Plot of observed response vs. predicted reponse and (B) normal probability plot of the studentized residuals.
Figure 7
Figure 7
Response surface 3D plots showing individual and interactive effects of variables on antibacterial activity of Streptomyces sp. JAJ13. (A) Effects of (NH4)2SO4 and CuSO4.5H2O on antibacterial activity. (B) Effects of K2HPO4 and CuSO4.5H2O on antibacterial activity. (C) Effects of K2HPO4 and (NH4)2SO4 on antibacterial activity.
Figure 8
Figure 8
HPLC profiles of secondary metabolites extracted from Streptomyces sp. JAJ13 culture broth under (A) unoptimized and (B) optimized levels of media components.
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