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The enzymatic hydrolysis of waste paper - a source of raw materials for production of liquid biofuels
Lepař, Petr ; Stloukal, Radek (referee) ; Gabriel, Petr (advisor)
In diploma thesis the process of enzymatic hydrolysis of waste paper as a source for the production of liquid biofuels is discused. It follows directly the homonymous diploma thesis from Ing. Brummer, and it is based on the findings, which were solved and decided in previous work. In the theoretical part there is a summarization of basic information on the enzymatic hydrolysis of waste paper and the associated influences of various factors of the rate and degree of hydrolysis. Higher attention is paid to a waste cardboard and its pretreatment methods due to the maximalization of the yield of hydrolysis. The next part summarizes options of the fermentative production of biofuels, focusing on the method of simultaneous saccharification and fermentation, where the further appropriate organism for ethanol fermentation is discussed. The last part is about the technological process from the raw material input to the separation of ethanol. In the experimental section the pre-treatment of waste paper in order to maximize the efficiency of hydrolysis was examined. The best results were achieved using a vibratory mill. In addition, various parameters for simultaneous saccharification and fermentation were optimized using enzymes from Novozymes® company and the yeast Saccharomyces cerevisiae. The conversion rate of waste paper cellulose to reducing sugars was observed by spectrophotometric method by Somogyi - Nelson and the amount of produced ethanol was quantified using HPLC / RI. As a part of this thesis some conditions (amount of enzyme, substrate, nutrients, yeasts, temperature, pH, type of buffer) were optimized to maximize the effectiveness of the overall process. All experiments were carried out on corrugated cardboard, which was chosen as the most promising material for hydrolysis that was among the waste paper pulp in diploma thesis by Ing. Brummer.
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Enzymatic hydrolysis of waste paper pulp - source of raw material for production of liquid biofuels
Brummer, Vladimír ; Ing.Marián Illáš (referee) ; Gabriel, Petr (advisor)
This master’s thesis is aimed at process of enzymatic hydrolysis of lignocellulosic material – waste paper as a source of raw material for production of liquid biofuels. In the theoretical part of this work are summarized previously used methods of hydrolysis and lignocellulosic materials used for the process of hydrolysis as a source of fermentable sugars for fermentation technology. The different types of waste paper are evaluated from the composition and usability with consideration to the papermaking process in order to select the appropriate type of waste paper for the enzymatic hydrolysis process. In the next part of this work are suggested technological premises and procedures for the preparation of raw materials and the subsequent enzymatic hydrolysis of these pre–treated materials. In the experimental part were optimized parameters of enzymatic hydrolysis using the Novozymes company enzyme package. Enzymatic degradation of cellulose to reducing sugars was observed using Somogyi – Nelson method. For the verification of hydrolysis conditions were used model materials with high cellulose content – pulp and filter paper. Conditions, which seems to be the best after testing on the model materials, were verified on specific waste paper materials – offset cardboard, recycled paper, matte MYsol paper and for comparison again on model materials – pulp and filter paper. The highest yields was achieved with the use of cardboard, which was further tested using various combinations of pretreatment to material for purpose of increase the yields of hydrolysis.
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Use of wastes from winemaking in food and cosmetics applications
Výmolová, Markéta ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The presented thesis deals with the various types of hydrolysis of waste from the production of wine, parings and bunch-stems for subsequent biotechnological use as enrichment of feeding mixtures and the production of bio-ethanol. A composite analysis was done at the start of the measurements for the determination of dry matter, extraction of oil from the parings and assessment of cellulose in the bunch-stems. As a part of this thesis a chemical and enzyme hydrolysis of waste materials have been performed. In hydrolysates the contents of reducing carbohydrates and antioxidants, polyphenols and flavonoids were analysed. The results showed that the samples with a concentration of 50 g/l of material hydrolysed in a water bath for 60 minutes were the best conditions for cultivating yeasts. Hydrolysed parings and bunch-stems were used as the production medium for the production of ethanol in Saccharomyces cerevisiae. A change of the concentration of carbohydrates and ethanol in the samples has been measured by HPLC. The sample hydrolysed enzymatically using cellulase exhibited the highest concentration of biomass (c = 2,140 g/l), the sample hydrolysed chemically in the water bath formed the highest content of ethanol (c = 9,880 g/l).
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Waste materials utilization for preparing hydrolysates for the fermentation phase.
Vadovičová, Natália ; Hrstka, Miroslav (referee) ; Babák, Libor (advisor)
Bachelor thesis focuses on the study and comparison of different types of hydrolysis, their optimization and maximization of yields for the upcoming fermentation. Orange peel was chosen as a substrate to conduct the experiments. First, the substrate was mechanically grinded to form a suspension. Each suspension then underwent one out of the examined methods of hydrolysis. Chosen methods were physical, such as microwaves, increased temperature or ultrasound, and chemical acidic and alkaline. Combinations of both types were also examined. The last optimized method was enzymatic hydrolysis. First set of experiments was conducted using enzymes Novozymes® NS50013 and NS50010. Production of cellulase and pectinase enzymes by A. niger during solid-state fermentation that lasted 10 days was also studied. The yields of reducing sugars of all the experiments were calculated using the Somogyi-Nelson method. Enzymatic hydrolysis was proven to be the most effective using the combination of both of the enzymes for a period of 96 hours at pH = 4.5 and temperature 45 °C. Yield of the reducing sugars under these conditions reached 27,4241 ± 0,0007 gl-1.
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Biotechnological production of polyhydroxyalkanoates using wastes of coffee production
Vašíčková, Monika ; Benešová, Pavla (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis is focused on study of production of polyhydroxyalkanoates (PHA) by bacteria Burkholderia cepacia and Burkholderia sacchari. Production of PHA has been studied using spent coffee grounds hydrolysates which have been prepared by different aproaches. In the thesis is compared standard method of coffee ground hydrolysis by mineral acid followed by enzymatic hydrolysis. Hydrolysates which have been produced were analyzed in terms of concentration of saccharides and efficiency of hydrolysis. The best producer of PHA (based on results) was bacterium Burkholderia cepacia, in the medium which has been prepared by hydrolysis of spent coffee grounds extracted by 60% solution of ethanol. Biomass yield of this cultivation was 3,553 g/l with 32,472% PHA content. This PHA contained 6,09% 3-hydroxyvalerate. In the other experiment, we verified alternative hydrolysis of spent coffee grounds – by commercially available enzymes. We used cellulase, hemicellulase and the enzymatic cocktail which has been produced by mould. Hemicellulase was the most effective hydrolytic enzyme and its application resulted in production of the highest amount of biomass – 5,708 g/l. In this cultivation, only homopolymer PHB has been which is probably caused by the fact, that during the enzymatic hydrolysis levulinic acid or any other potential precursor of 3HV is formed.
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Production of liquid biofuels by using enzymatic hydrolysis of waste paper
Roštek, Martin ; Jecha, David (referee) ; Brummer, Vladimír (advisor)
This bachelor thesis focuses on the retrieval summary of current knowledge about the process of enzymatic hydrolysis with utilization of waste paper as raw material, in order to convert it into liquid biofuels. The thesis summarizes the general knowledge of the lignocellulosic a raw materials and their resources, an overview of available hydrolysis methods for materials and fermentation technologies, and also summary of methods currently used to rise enzymatic hydrolysis process effectivity. The next section provides an overview of commercial plants which use enzymatic hydrolysis of lignocellulosic materials.
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Enzymatic hydrolysis of waste cardboard using the SSF method - a source of raw materials for the production of liquid biofuels.
Hlaváček, Viliam ; Stloukal, Radek (referee) ; Gabriel, Petr (advisor)
This master’s thesis discusses the useof enzymatic hydrolysis process of waste cardboard using simultaneous saccharification and fermentation (SSF) as a source of raw materials for production of liquid biofuels. This thesis is based on theses written by Ing. Brummer and Ing.Lepař.Thus, results gained in these works have been used and also further developed. The theoretical part summarizes the reasons for further development of SSF method and discusses, as well, the achievements reached in the processing of lignocellulosic waste materials by the SSF method so far.This section also discusses the general characteristics of lignocellulosic materials and also of the cellulolytic enzymes. It focusses also on individual pretreatment methods of lignocellulosic material and options of increasing the yield of the whole process. The experimental part verifies the particular results reached in previous theses and at the same time a further optimization of the method has been carried out because of the transfer of the whole process into a fermenter. Cardboard was set as the substrate for the experiments as it was evaluated by Ing. Brummer as the best one for enzymatic hydrolysis which was carried out by enzymes from Novozymes®. Parameters such as temperature, pH and kind of used buffer, the loading concentration of substrate and enzymes, were set according to the thesis of Ing. Lepař, which was aimed to their optimization. The SSF process done in fermenter of 2.0 l volume confirmed the previous results and furthermore it has been more effective through optimization of the added inoculum volume. It has been confirmed that the best substrate is cardboard finely grinded by vibrating mill. Also experiments with added nutrients had been done as an effort to increase the ethanol concentration, but these haven’t resulted insatisfying results. The maximal concentration of ethanol was 23,49 g/l, which was achieved after further optimization of various conditions. This result equals to experimental yield of 84,79 %.
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Study of growth and optimization of selected metabolites production by Zymomonas mobilis
Lukačková, Adéla ; Vránová, Dana (referee) ; Babák, Libor (advisor)
In the diploma thesis are discussed the process of enzymatic hydrolysis of waste paper as a source for the production of bioethanol by bacteria Zymomonas mobilis. In the theoretical part summarize basic information about particular methods of hydrolysis, about paper used as a raw material for enzymatic hydrolysis, about possibilities of the fermentative production of bioethanol focusing on the method of simultaneous saccharification and fermentation comparison with enzymatic hydrolysis and fermentation. Suitable microorganisms for ethanolic fermentation and simultaneous saccharification and fermentation and their advantages and disadvantages, are further discussed in this part as well. The theoretical part ends with the suggestion of the technological process for production of bioetanol. It covers all necessary steps from the input of raw material to the separation of produced ethanol. In the experimental part various parameters of hydrolysis, fermentation and simultaneous saccharification and fermentation were optimized using enzymes from Novozymes® company and the Zymomonas mobilis CCM2770 and Zymomonas mobilis LMG457 bacterium. The conversion rate of paper cellulose to gluckose and production of ethanol were observed by HPLC/RI method. Type of buffer, quantity of cells, enzyme and substrate were optimized in order to maximize the efficiency of the process. All experiments were performed on paper containing high amount of cellulose and for comparison on standard medium which contains gluckose. The highest yields was achieved with the use of Novozymes® Cellulosic ethanol enzyme Kit. The strain Zymomonas mobilis LMG457 has demonstrated as a better producer.
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Use of technical hemp extracts in hair cosmetics
Benková, Sarah ; Holub, Jiří (referee) ; Márová, Ivana (advisor)
Technical hemp as a versatile plant has found applications in many sectors of the industry. In cosmetics, it is mainly used for bioactive substances as well as proteins. The subject of this bachelor thesis was the preparation and characterization of a fraction containing hydrolysed hemp proteins and its use in hair cosmetics. In the theoretical part, the characteristics of the plant as well as its chemical composition are processed. Attention is paid especially to hemp proteins, their hydrolysis possibilities, and the subsequent use of hydrolysates in hair cosmetics. Within the experimental part, the preparation of hemp protein hydrolysates from hempseed cake of the Santhica 70 variety was optimized using the industrial enzymes TS-E 1930 and TS-E 1993 and the subsequent determination of the protein content and profile of prepared hydrolysates. The carbohydrate, chlorophyll, phenolic and flavonoid content of the analysed hydrolysates was also determined. A hair conditioner was prepared from the selected hydrolysate and tested by ten respondents. Thanks to the prepared conditioner, hair cosmetics with the addition of hydrolysed hemp proteins has been shown to enhance shine and pliability of the hair, as well as help the healing of frayed ends.
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