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Preparation and use of acid proteases for digestion in H/D exchange.
Kukla, Jan ; Man, Petr (advisor) ; Pompach, Petr (referee)
- 5 - Abstract Hydrogen/deuterium exchange coupled to mass spectrometry (HX-MS) utilizes the spontaneous exchange of protein backbone amide hydrogens for deuterium atoms from solution to gain information about changes in protein structure. To localize these changes to specific areas of the protein, enzymatic digestion by aspartate proteases is used. The proteases' ability to produce small overlapping peptides and to provide full sequence coverage of the studied protein is essential for pinpointing the protein regions of interest. In this study recombinant proteases nepenthesin I (Nepenthes gracilis) and rhizopuspepsin (Rhizopus chinensis) were prepared and compared to commercially available proteases porcine pepsin A and aspergillopepsin (Aspergillus saitoi). The comparison was performed using various activity assays, where the effects of pH, temperature and denaturing and reducing agents on the activity of the proteases were studied. All four proteases were also immobilized on a polymeric resin POROS and their activity in an online HX-MS digestion setup was tested using myoglobin as a model substrate.
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Konstrukční řešení tenzometrické měřicí hlavy
Kukla, Jan
The main objective of my thesis is to design the constuction of strain gauge mea-surring head. This constrction allows testing float in the pool and facilitate the job of a human operator. By measuring we determined the physical properties of the float. The construction is mounted on the robot KUKA. I chose suitable sensor single-point for measuring buoyacy and tensile strength float on the surface. Strain sensor has an cover IP67. Material of the supportng structure is made of aluminium alloy from company MayTec. The sensors are located in the axis of the center of gravity of the float, which i experimentally calculated. From my experience, character of the float in work on the surface and under load be expected change center of gravity. Construction is adapted for this situation. Final product is light and durable construction with precise sensors and low price.
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Preparation and use of acid proteases for digestion in H/D exchange.
Kukla, Jan ; Man, Petr (advisor) ; Pompach, Petr (referee)
- 5 - Abstract Hydrogen/deuterium exchange coupled to mass spectrometry (HX-MS) utilizes the spontaneous exchange of protein backbone amide hydrogens for deuterium atoms from solution to gain information about changes in protein structure. To localize these changes to specific areas of the protein, enzymatic digestion by aspartate proteases is used. The proteases' ability to produce small overlapping peptides and to provide full sequence coverage of the studied protein is essential for pinpointing the protein regions of interest. In this study recombinant proteases nepenthesin I (Nepenthes gracilis) and rhizopuspepsin (Rhizopus chinensis) were prepared and compared to commercially available proteases porcine pepsin A and aspergillopepsin (Aspergillus saitoi). The comparison was performed using various activity assays, where the effects of pH, temperature and denaturing and reducing agents on the activity of the proteases were studied. All four proteases were also immobilized on a polymeric resin POROS and their activity in an online HX-MS digestion setup was tested using myoglobin as a model substrate.
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Výroba a vlastnosti vrtáků
Kukla, Jan
The bachelor work is based on a survey of the production and basic characteristic of the production and properties of drills in the company, which deals with the total production of drilling tools. Five-axis CNC tool grinders are able to achieve high precision processing with a wide range of different types of materials. Hardness, toughness and heat resistance of drill bits are specified prefabricated bore material (aluminium, stainless steel, etc.), the geometry of a cutting edge drills (Mitsubishi, Gühring, "RDS", etc.) and the type of the coating (TiAlN, AlCrN, ArTiN, CC Alu speed, etc.), or processing of the surface (nitriding, blackening).
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Marketing strategy of a particular company
Votavová, Alena ; Průša, Přemysl (advisor) ; Kukla, Jan (referee)
This thesis deals with the marketing strategy of a company F AIR spol. s r. o.,which is the first flying school in the Czech Republic and has its main domicile at the airport Benešov. The objective of this thesis is to introduce the products of the company, to analyse its marketing strategy and marketing environment, and finally to propose feasible changes of the contemporary strategy. To achieve this objective, I have applied several subanalyses and my own marketing research. The accumulated figures are evaluated in the final SWOT analysis. I have used this figures to bring up several changes, which could be conductive to the increase of the company's turnover and to the improvment of its customer's satisfaction.
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