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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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Biotechnological production of polyhydroxyalkanoates on wheat bran
Guziurová, Pavlína ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
The bachelor thesis deals with the biotechnological production of polyhydroxyalkanoates (PHA) from lignocellulosic waste – wheat bran. The aim was to determine whether the hydrolysate from wheat bran subjected to different types of pretreatment could be used as a substrate for the production of PHA using selected bacterial strains. The selected strains were Halomonas organivorans, Halomonas halophila and two strains of Schlegelella thermodepolymerans. Halophilic strains were proved as the best producers and were subsequently used for cultivations. The hydrolysates after neutral pretreatment were utilized by the bacteria most efficiently, due to the lowest content of microbial inhibitors (phenolic substances) where the bacteria produced the most PHA. The highest value of produced PHA was determined on the hydrolysate after neutral pretreatment by using the strain Halomonas organivorans, namely 2,82 g/l. The hydrolysate was also used for the production of lactic acid bacterial strains of Lactobacillus. The highest achieved concentration of produced lactic acid was 16,73 g/l by the Lactobacillus casei strain.
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Biotechnological production of polyhydroxyalkanoates on wheat bran
Guziurová, Pavlína ; Pernicová, Iva (referee) ; Obruča, Stanislav (advisor)
The bachelor thesis deals with the biotechnological production of polyhydroxyalkanoates (PHA) from lignocellulosic waste – wheat bran. The aim was to determine whether the hydrolysate from wheat bran subjected to different types of pretreatment could be used as a substrate for the production of PHA using selected bacterial strains. The selected strains were Halomonas organivorans, Halomonas halophila and two strains of Schlegelella thermodepolymerans. Halophilic strains were proved as the best producers and were subsequently used for cultivations. The hydrolysates after neutral pretreatment were utilized by the bacteria most efficiently, due to the lowest content of microbial inhibitors (phenolic substances) where the bacteria produced the most PHA. The highest value of produced PHA was determined on the hydrolysate after neutral pretreatment by using the strain Halomonas organivorans, namely 2,82 g/l. The hydrolysate was also used for the production of lactic acid bacterial strains of Lactobacillus. The highest achieved concentration of produced lactic acid was 16,73 g/l by the Lactobacillus casei strain.
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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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