About: Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris.

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About: Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. Goto Sponge NotDistinct Permalink

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	Heterologous expression of the bacterial enzyme haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26 in methylotrophic yeast Pichia pastoris is reported. The haloalkane dehalogenase gene linB was subcloned into the pPICZaA vector and integrated into the genome of P. pastoris. The recombinant LinB secreted from the yeast was purified to homogeneity and biochemically characterized. The deglycosylation experiment and mass spectrometry measurements showed that the recombinant LinB expressed in P. pastoris is glycosylated with a 2.8 kDa size of high mannose core. The specific activity of the glycosylated LinB was 15.6 +/- 3.7 micromol/min/mg of protein with 1,2-dibromoethane and 1.86 +/- 0.36 micromol/min/mg of protein with 1-chlorobutane. Activity and solution structure of the protein produced in P. pastoris is comparable with that of recombinant LinB expressed in Escherichia coli. The melting temperature determined by the circular dichroism (41.7 +/- 0.3C for LinB expressed in P. Heterologous expression of the bacterial enzyme haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26 in methylotrophic yeast Pichia pastoris is reported. The haloalkane dehalogenase gene linB was subcloned into the pPICZaA vector and integrated into the genome of P. pastoris. The recombinant LinB secreted from the yeast was purified to homogeneity and biochemically characterized. The deglycosylation experiment and mass spectrometry measurements showed that the recombinant LinB expressed in P. pastoris is glycosylated with a 2.8 kDa size of high mannose core. The specific activity of the glycosylated LinB was 15.6 +/- 3.7 micromol/min/mg of protein with 1,2-dibromoethane and 1.86 +/- 0.36 micromol/min/mg of protein with 1-chlorobutane. Activity and solution structure of the protein produced in P. pastoris is comparable with that of recombinant LinB expressed in Escherichia coli. The melting temperature determined by the circular dichroism (41.7 +/- 0.3C for LinB expressed in P. (en)
Title	Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. (en)
skos:prefLabel	Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. (en)
skos:notation	RIV/00216224:14310/06:00017437!RIV10-MSM-14310___
http://linked.open...avai/riv/aktivita	Z
http://linked.open...avai/riv/aktivity	Z(MSM0021622412), Z(MSM0021622415)
http://linked.open...iv/cisloPeriodika	46
http://linked.open...vai/riv/dodaniDat	2010
http://linked.open...aciTvurceVysledku	Prokop, Zbyněk Damborský, Jiří Zdráhal, Zbyněk Chaloupková, Radka Monincová, Marta
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	Masarykova univerzita / Přírodovědecká fakulta
http://linked.open...dnocenehoVysledku	475335
http://linked.open...ai/riv/idVysledku	RIV/00216224:14310/06:00017437
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Dehalogenation; Pichia pastoris; Heterologous expression; Glycosylation; Thermal stability; Catalytic activity (en)
http://linked.open.../riv/klicoveSlovo	Catalytic activity Thermal stability Glycosylation Heterologous expression Pichia pastoris Dehalogenation
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[1A5EAB386BA2]
http://linked.open...i/riv/nazevZdroje	PROTEIN EXPRESSION AND PURIFICATION
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...UplatneniVysledku	2006
http://linked.open...v/svazekPeriodika	1/2006
http://linked.open...iv/tvurceVysledku	Chaloupková, Radka Damborský, Jiří Nagata, Yuji Prokop, Zbyněk Zdráhal, Zbyněk Monincová, Marta Nakamura, Takashi Zamocky, Marcel
http://linked.open...ain/vavai/riv/wos	000235902700010
http://linked.open...n/vavai/riv/zamer	Interactions among the chemicals, environment and biological systems and their consequences on the global, regional and local scales (INCHEMBIOL) Molecular basis of cell and tissue regulations
issn	1046-5928
number of pages	7 (xsd:int)
http://localhost/t...ganizacniJednotka	14310
is http://linked.open...avai/riv/vysledek of	Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris. Expression of Glycosylated Haloalkane Dehalogenase LinB in Pichia pastoris.

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