J Vet Sci.  2010 Dec;11(4):327-332. 10.4142/jvs.2010.11.4.327.

Optimization of culture media of pathogenic Mycoplasma hyopneumoniae by a response surface methodology

• Hwang MH
• Damte D
• Cho MH
• Kim YH
• Park SC

• KMID: 1072182
• DOI: http://doi.org/10.4142/jvs.2010.11.4.327

Abstract

Composition of culture medium for mass production of Mycoplasma hyopneumoniae was optimized using a response surface methodology (RSM). Initially, the influence of glucose, thallium acetate, fresh yeast extract, horse serum, and porcine serum on the production of mycoplasmal protein was assessed using a 'one factor at a time' technique. Next, factors with a significant effect, including fresh yeast extract, and horse and porcine sera, were selected for further optimization using a central composite design (CCD) of RSM. The experimental results were fitted into a second order polynomial model equation. Estimated optimal condition of the factors for maximum production of mycoplasmal protein (i.e., triple-fold increase from 0.8 mg/L produced by basal mycoplasma media to 2.5 mg/L) was 10.9% fresh yeast extract, 15% horse serum, and 31.5% porcine serum (v/v). For the optimized conditions, a 2.96 mg/L experimental result was observed, similar to the estimated optimal conditions result of the CCD.

Keyword

fresh yeast extract; horse serum; Mycoplasma hyopneumoniae; optimization; response surface analysis

MeSH Terms

Biotechnology/*methods
Culture Media/*chemistry
Mycoplasma hyopneumoniae/*growth & development

Figure

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  Fig. 1 Effect of five experimental factors on mycoplasmal protein production (A) 50% glucose solution, (B) horse serum, (C) fresh yeast extract, (D) porcine serum, and (E) 2% thallium acetate solution each varying at five levels while all the other factors were kept constant. *p < 0.05, BM: base medium.

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  Fig. 2 Response surface plot showing the effect of the three variables/experimental factors on mycoplasmal protein yield; (A) Effect of horse serum and porcine serum, (B) effect of yeast extract and porcine serum, and (C) effect of horse serum and yeast extract.

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  Fig. 3 Comparison of the base media (BM), and modified and optimized Friis media on mycoplasmal protein production at different time intervals. *p < 0.001, **p < 0.01.

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