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Polyhydroxyalkanoates as metabolites of prokaryotic microorganisms adapted to extreme conditions
Vodička, Juraj ; Slaninová, Eva (referee) ; Obruča, Stanislav (advisor)
The objective of this work is a study of polyhydroxyalkanoates as bacterial carbon and energy storage forms, which are produced intracellularly. The theoretical part of the study is focused on physical properties of PHA, their biotechnological production and degradation. The experimental part deals with production of polyhydroxyalkanoates by halophile bacterium Halomonas organivorans using several different carbon substrates. Firstly, the gene encoding for PHA-synthase, phaC, was successfully amplified and detected. Extracellular lipolytical activity was also proved. Secondly, the most appropriate substrate for bacterial growth and PHA production were found to be hydrocarbons, especially galactose and mannose. Due to these results, it is highly desirable to use coffee grounds for the biotechnological production of PHB, as galactomannan-rich waste. On the other hand, lactose and xylose have shown to be unsuitable carbon sources. For that reason, waste whey and lignocellulosic materials rich for pentoses are inconvenient substrates for the intended purpose. There is a potential use of waste molasses because of good results of sucrose at contrast. By the cultivation on kitchen waste and waste glycerol were the growth slow and the production of PHA reduced significantly. The optimal salt concentration for bacterial growth and PHA production were discovered as 60 grams per litre. Addition of several precursors of 3-hydroxyvalerate after 24 hours of cultivation have not induced the copolymer P(3HB-co-3HV) formation. Even though these precursors have inhibited the bacterial growth, the significant increase of the PHB percent contend was observed. In general, the yield of the production has been considerable, it should be noted than almost in each successful cultivation the PHA content in bacterial cells exceeded 50 weight %. With a respect to results, more experiments for full evaluation of H. organivorans as a potential biotechnological PHA producing bacteria are regarded.
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Polyhydroxyalkanoates as metabolites of prokaryotic microorganisms adapted to extreme conditions
Vodička, Juraj ; Slaninová, Eva (referee) ; Obruča, Stanislav (advisor)
The objective of this work is a study of polyhydroxyalkanoates as bacterial carbon and energy storage forms, which are produced intracellularly. The theoretical part of the study is focused on physical properties of PHA, their biotechnological production and degradation. The experimental part deals with production of polyhydroxyalkanoates by halophile bacterium Halomonas organivorans using several different carbon substrates. Firstly, the gene encoding for PHA-synthase, phaC, was successfully amplified and detected. Extracellular lipolytical activity was also proved. Secondly, the most appropriate substrate for bacterial growth and PHA production were found to be hydrocarbons, especially galactose and mannose. Due to these results, it is highly desirable to use coffee grounds for the biotechnological production of PHB, as galactomannan-rich waste. On the other hand, lactose and xylose have shown to be unsuitable carbon sources. For that reason, waste whey and lignocellulosic materials rich for pentoses are inconvenient substrates for the intended purpose. There is a potential use of waste molasses because of good results of sucrose at contrast. By the cultivation on kitchen waste and waste glycerol were the growth slow and the production of PHA reduced significantly. The optimal salt concentration for bacterial growth and PHA production were discovered as 60 grams per litre. Addition of several precursors of 3-hydroxyvalerate after 24 hours of cultivation have not induced the copolymer P(3HB-co-3HV) formation. Even though these precursors have inhibited the bacterial growth, the significant increase of the PHB percent contend was observed. In general, the yield of the production has been considerable, it should be noted than almost in each successful cultivation the PHA content in bacterial cells exceeded 50 weight %. With a respect to results, more experiments for full evaluation of H. organivorans as a potential biotechnological PHA producing bacteria are regarded.
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