Proteolytické enzymy vegetativních forem a spor bakterie Paenibacillus larvae

Abstract

Due to the high resistance of the spores, the bacterium Paenibacillus larvae is the most dangerous bacterial pathogen of the honey bee (Apis mellifera). Thanks to its biological properties and restricted pathogenicity, this bacterium can be used as a model organism to study gram positive sporulating aerobic rods. This work is focused on completing information about secreted proteases of this bacterium and in a study of proteases bound in a spore structure. MYPGP medium was used for the cultivation of P. larvae. In this medium, lysis of the culture was shown after 40 hours of cultivation. The pH of the medium decreased below 6.4 by lysis. The induction of temperate bacteriophage BLA was detected as a causative agent of this lysis. A new sporulation medium called HCBB agar was proposed for the sporulation of P. larvae. In comparison with HCBB agar with MYPGP agar by 31 strains of P. larvae stored in our collection, HCBB agar was evaluated as an appropriate sporulation medium with a median of sporulatin 4.2 ' 106 spores per cm2 in aerobic conditions and 5.65 ' 106 spores per cm2 in aerobic conditions with 10 % CO2. For purification of the secreted proteases, a one-day culture incubated at room temperature was used. Optimal purification of 87/74 kDa and 42/40 kDa proteases was observed after application of this...

Univerzita Karlova v Praze Lékařská fakulta v Plzni Ústav mikrobiologie Proteolytické enzymy vegetativních forem a spor bakterie Paenibacillus larvae (Dizertační práce) Jaroslav Hrabák Plzeň, 2007 Jaroslav Hrabák Dizertační práce 1 Název: Proteolytické enzymy vegetativních forem a spor bakterie Paenibacillus larvae (Proteolytic Enzymes of Vegetative Forms and Spores of the Bacterium Paenibacillus larvae) Autor: Ing. Jaroslav Hrabák Pracoviště: Ústav mikrobiologie Lékařská fakulta v Plzni Univerzita Karlova v Praze Adresa fakulty: Husova 3, 305 06 Plzeň Adresa pracoviště: Dr. E. Beneše 13, 305 99 Plzeň Školitel: RNDr. Karel Martínek, Ph.D. Studijní program: Všeobecné lékařství Obor: Lékařská mikrobiologie Rok vydání: 2007 Místo vydání: Plzeň Počet číslovaných stran: 94 Počet obrázků: 33 Počet tabulek: 5 Počet příloh: 0 Počet citovaných pramenů: 75 Jaroslav Hrabák Dizertační práce 2 Obsah Obsah…………………………………………………………………………………….. 2 Prohlášení………………………………………………………………………………… 6 Předmluva………………………………………………………………………………... 7 Souhrn……………………………………………………………………………………. 8 Summary…………………………………………………………………………………..10 1. Úvod…………………………………………………………………………………… 12 1.1. Taxonomické zařazení bakterie Paenibacillus larvae………………………..12 1.2. Patogeneze moru včelího plodu……………………………………………....14 1.3. Fyziologie bakterie P. larvae s...