NDLI: Gene cloning and expression in Escherichia coli of a bi-functional β-d-xylosidase/α-l-arabinosidase from Sulfolobus solfataricus involved in xylan degradation

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Gene cloning and expression in Escherichia coli of a bi-functional β-d-xylosidase/α-l-arabinosidase from Sulfolobus solfataricus involved in xylan degradation

Content Provider	Springer Nature Link
Author	Morana, Alessandra Paris, Ornella Maurelli, Luisa Rossi, Mosé Cannio, Raffaele
Copyright Year	2006
Abstract	An open reading frame encoding a putative bi-functional β-d-xylosidase/α-l-arabinosidase (Sso3032) was identified on the genome sequence of Sulfolobus solfataricus P2, the predicted gene product showing high amino-acid sequence similarity to bacterial and eukaryal individual β-d-xylosidases and α-l-arabinosidases as well as bi-functional enzymes such as the protein from Thermoanaerobacter ethanolicus and barley. The sequence was PCR amplified from genomic DNA of S. solfataricus P2 and heterologous gene expression obtained in Escherichia coli, under optimal conditions for overproduction. Specific assays performed at 75°C revealed the presence in the transformed E. coli cell extracts of this archaeal activity involved in sugar hydrolysis and specific for both substrates. The recombinant protein was purified by thermal precipitation of the host proteins and ethanol fractionation and other properties, such as high thermal activity and thermostability could be determined. The protein showed a homo-tetrameric structure with a subunit of molecular mass of 82.0 kDa which was in perfect agreement with that deduced from the cloned gene. Northern blot analysis of the xarS gene indicates that it is specifically induced by xylan and repressed by monosaccharides like d-glucose and l-arabinose.
Starting Page	123
Ending Page	132
Page Count	10
File Format	PDF
ISSN	14310651
Journal	Extremophiles
Volume Number	11
Issue Number	1
e-ISSN	14334909
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-10-11
Publisher Place	Tokyo
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	β-d-xylosidase/α-l-arabinosidase Sulfolobus solfataricus Xylan degradation Gene cloning and expression Ecology Biotechnology Microbiology
Content Type	Text
Resource Type	Article
Subject	Medicine Molecular Medicine Microbiology

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