About: Serine protease from midgut of Bombus terrestris males

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	A serine protease was isolated from midguts of the bumblebee male Bombus terrestris by a combination of precipitation procedures with column chromatography. The purified enzyme exhibited two bands with molecular masses of 25 and 26 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These bands showed a proteolytic activity in zymography assay. Midgut enzymes showed optimum proteolytic activity at pH 9 and 35 degrees C using N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenyl-alanine 4-nitroanilide as a substrate. The Michaelis constant (Km) and maximum reaction rate (Vmax) were 0.55 +/- 0.042 mM and 0.714 +/- 0.056 mol p-nitroalanine produced min1 mg protein1, respectively. Inhibition was affected by trypsin inhibitor, but not by phenylmethylsulfonyl fluoride and N-tosyl-L-phenylalanine chloromethyl ketone, which indicated the trypsin-like but not chymotrypsin-like specificity. The identity of the serine protease was confirmed by nanoliquid-tandem mass spectrometry. Eleven unique peptides of the B. terrestris serine protease were found. It shows high homology to a previously reported B. ignitus serine protease covering more than 65% of the protein amino acid sequence. A serine protease was isolated from midguts of the bumblebee male Bombus terrestris by a combination of precipitation procedures with column chromatography. The purified enzyme exhibited two bands with molecular masses of 25 and 26 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These bands showed a proteolytic activity in zymography assay. Midgut enzymes showed optimum proteolytic activity at pH 9 and 35 degrees C using N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenyl-alanine 4-nitroanilide as a substrate. The Michaelis constant (Km) and maximum reaction rate (Vmax) were 0.55 +/- 0.042 mM and 0.714 +/- 0.056 mol p-nitroalanine produced min1 mg protein1, respectively. Inhibition was affected by trypsin inhibitor, but not by phenylmethylsulfonyl fluoride and N-tosyl-L-phenylalanine chloromethyl ketone, which indicated the trypsin-like but not chymotrypsin-like specificity. The identity of the serine protease was confirmed by nanoliquid-tandem mass spectrometry. Eleven unique peptides of the B. terrestris serine protease were found. It shows high homology to a previously reported B. ignitus serine protease covering more than 65% of the protein amino acid sequence. (en)
Title	Serine protease from midgut of Bombus terrestris males Serine protease from midgut of Bombus terrestris males (en)
skos:prefLabel	Serine protease from midgut of Bombus terrestris males Serine protease from midgut of Bombus terrestris males (en)
skos:notation	RIV/61388963:_____/13:00391467!RIV14-TA0-61388963
http://linked.open...avai/riv/aktivita	I P
http://linked.open...avai/riv/aktivity	I, P(GA203/09/1446), P(TA01020969)
http://linked.open...iv/cisloPeriodika	3
http://linked.open...vai/riv/dodaniDat	2014
http://linked.open...aciTvurceVysledku	Kindl, Jiří Pichová, Iva Valterová, Irena Brabcová, Jana Zarevúcka, Marie
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Ústav organické chemie a biochemie AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	104762
http://linked.open...ai/riv/idVysledku	RIV/61388963:_____/13:00391467
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Bombus terrestris; midgut; serine protease; bumblebee (en)
http://linked.open.../riv/klicoveSlovo	serine protease midgut bumblebee Bombus terrestris
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[B90277AD89EC]
http://linked.open...i/riv/nazevZdroje	Archives of Insect Biochemistry and Physiology
http://linked.open...in/vavai/riv/obor	CE
http://linked.open...ichTvurcuVysledku	5 (xsd:int)
http://linked.open...cetTvurcuVysledku	8 (xsd:int)
http://linked.open...vavai/riv/projekt	Study of Enzymes Involved in the Biosynthesis of Pheromones of Model Species of Bumblebees Pollinators as a crucial factor in agriculture.
http://linked.open...UplatneniVysledku	2013
http://linked.open...v/svazekPeriodika	82
http://linked.open...iv/tvurceVysledku	Kindl, Jiří Pichová, Iva Valterová, Irena Brabcová, Jana Mikšík, Ivan Zarevúcka, Marie Jágr, Michal Brabcová, J.
http://linked.open...ain/vavai/riv/wos	000315102100002
issn	0739-4462
number of pages	12 (xsd:int)
http://bibframe.org/vocab/doi	10.1002/arch.21075

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