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Growth screening of Rhodospirillum rubrum using microcalorimetry
Vetchý, Antonín ; Slaninová, Eva (referee) ; Krouská, Jitka (advisor)
The presented work deals with the study of the growth of the bacterium Rhodospirillum rubrum using microcalorimetry. The theoretical part is devoted to the characterization of the investigated microorganism and the description of the main measurement method. Two strains of the mentioned bacterium were selected for the measurement – the wild type strain and the mutant strain with a deletion of the gene for PHA synthase. Firstly, the optimization of measurement conditions was performed, including the examination of various values of the initial optical density. The main objective of the work was to compare the growth of both strains of R. rubrum on different substrates differing in carbon source. Normalized heat flow curves obtained by microcalorimetry, and growth curves measured by the traditional spectrophotometric method were used for comparison. In conclusion, replicable measurement conditions were established, providing comparable results for both strains of bacteria. The most suitable medium for biomass growth of both strains of the bacterium Rhodospirillum rubrum appears to be the SYN FAY medium, while the least suitable medium seems to be the SYN AY medium containing acetate, where partial inhibitory effects on bacterial growth were observed. The results of the analysis using the microcalorimeter were further confirmed and extended by measuring growth curves on an ELISA reader.
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Behaviour of microorganisms using microcalorimetry
Piletskaya, Maryia ; Sedláček, Petr (referee) ; Krouská, Jitka (advisor)
The diploma thesis deals with the study of the behavior of a microorganism with the help of microcalorimetry. The theoretical part of the thesis deals mainly with the characterization of the microorganism Rhodospirillum rubrum selected for experimental work and the application of microcalorimetry in microbiology. The second goal was to screen the growth of the bacterial strain R. rubrum using different substrates, and subsequently, based on experimental results, to select the most suitable substrate, and optimize its concentration and aeration of the medium for the microorganism using microcalorimetry. Hexanoate, malate, acetate and fructose were used to examine the most suitable substrate by microcalorimetry. Another goal was data processing and discussion of the advantages and disadvantages of substrate selection and media aeration with respect to bacterial strain growth. Hexanoate was found to be the best substrate for growth, followed by optimization of the concentration and aeration of the medium. The results of the microcalorimetric measurement were confirmed by spectrophotometric study of the growth curve of the investigated bacteria. The effect of osmotic pressure on bacteria and the dependence of their survival on the amount of polyhydroxyalkanoates (PHA) in the cell were also studied. The amount of PHA in the biomass was determined chromatographically.
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Behaviour of microorganisms using microcalorimetry
Piletskaya, Maryia ; Sedláček, Petr (referee) ; Krouská, Jitka (advisor)
The diploma thesis deals with the study of the behavior of a microorganism with the help of microcalorimetry. The theoretical part of the thesis deals mainly with the characterization of the microorganism Rhodospirillum rubrum selected for experimental work and the application of microcalorimetry in microbiology. The second goal was to screen the growth of the bacterial strain R. rubrum using different substrates, and subsequently, based on experimental results, to select the most suitable substrate, and optimize its concentration and aeration of the medium for the microorganism using microcalorimetry. Hexanoate, malate, acetate and fructose were used to examine the most suitable substrate by microcalorimetry. Another goal was data processing and discussion of the advantages and disadvantages of substrate selection and media aeration with respect to bacterial strain growth. Hexanoate was found to be the best substrate for growth, followed by optimization of the concentration and aeration of the medium. The results of the microcalorimetric measurement were confirmed by spectrophotometric study of the growth curve of the investigated bacteria. The effect of osmotic pressure on bacteria and the dependence of their survival on the amount of polyhydroxyalkanoates (PHA) in the cell were also studied. The amount of PHA in the biomass was determined chromatographically.
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