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. 2020 Oct 23;10(1):190.
doi: 10.1186/s13568-020-01127-y.

Design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer

Dayana Morales-Borrell  1 , Nemecio González-Fernández  2 , Néstor Mora-González  2   3 , Carlos Pérez-Heredia  2   4 , Ana Campal-Espinosa  2 , Eddy Bover-Fuentes  2 , Eladio Salazar-Gómez  2   5 , Yissel Morales-Espinosa  2
Affiliations

Design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer

Dayana Morales-Borrell et al. AMB Express. .

Abstract

Culture medium composition is one of the most important parameters to analyze in biotechnological processes with industrial purposes. The aim of this study was to design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer. The influence of several carbon and nitrogen sources and their molar ratios on P. indica H32 growth was investigated. The effect of different micronutrients such as mineral salts and vitamin on P. indica H32 growth was determined as well. A mixture design based on Design-Expert 10.0 Software was performed to optimize the culture medium concentration. Finally, in the designed medium, an attribute of the biological mechanism of action of the P. indica H32 against nematodes, was evaluated: the hydrogen sulfide production. It was found that tested carbon/nitrogen ratios were not a significant influence on P. indica H32 growth. Growth of P. indica H32 was favored with use of sucrose, yeast extract and phosphate buffer without the addition of any tested micronutrients. An optimal concentration of 10 g/L sucrose and 5 g/L yeast extract were obtained at a cost of 0.10 $/L. In this concentration, the specific growth rate (µ) and maximal optical density (Xmax) were equal to 0.439 h- 1 and 8.00 respectively. It was evidenced that under the culture conditions used, P. indica H32 produced hydrogen sulfide. The designed medium led to a 1.08 $/L reduction of costs in comparison to LB medium. These results were critical to carry on with biotechnological development of P. indica H32 as a bioproduct.

Keywords: Bioproduct; Culture media; Mixture design; Nematicidal activity; Optimization; Pseudoxanthomonas indica.

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Conflict of interest statement

The authors declares that they have no competing interests.

Figures

Fig. 1
Fig. 1
Influence of carbon and nitrogen sources on µ and Xmax of P. indica H32. a Values of µ obtained with the evaluated carbon sources. b Values of µ obtained with the evaluated nitrogen sources. c Values of Xmax obtained with the evaluated nitrogen sources. G: glucose. S: sucrose. AC: ammonium chloride. BP: bacteriological peptone. YE: yeast extract. All bars represent the mean values ± 95.0 percent LSD intervals
Fig. 2
Fig. 2
Influence of mineral salts and vitamins on P. indica H32 growth. a Values of µ obtained with the tested media. b Values of Xmax obtained with the tested media. 1: medium 1 (7.73 g/L sucrose, 7.07 g/L yeast extract and 100 mL/L phosphate buffer). 2: medium 1 + CaCl2. 3: medium 1 + MgSO4. 4: medium 1 + NaCl. 5: medium 1 + trace salts solution. 6: medium 1 + vitamins. 7: medium 1 + CaCl2 + MgSO4 + NaCl + trace salts solution + vitamins. All bars represent the mean values ± 95.0 percent LSD intervals
Fig. 3
Fig. 3
Predicted optimum of the response variables from the optimization of the culture medium (H medium) concentration using the two-component mixture. A: Sucrose. B: Yeast extract. All experimental points of the design contain the same proportion of phosphate buffer (100 mL/L). formula image Design points
Fig. 4
Fig. 4
Hydrogen sulfide production by the P. indica H32 at 24 h of culture in the H medium. 1, 2 and 3 number of replicas. A: P. indica H32. B: Negative control (H medium). C: Positive control (Brevibacterium celere C924, active component of the HeberNem® registered biological product for the nematode control and plant growth promoting)
See this image and copyright information in PMC

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