Cloning and production of Xylanase B fromPaenibacillus barcinonensisinBacillus subtilishosts

Autor:	Oscar Gallardo, F. I. Javier Pastor, Pilar Diaz
Rok vydání:	2007
Předmět:	Protease Bacillaceae biology Chemistry medicine.medical_treatment Bacillus subtilis biology.organism_classification medicine.disease_cause Biochemistry Bacillales Catalysis Microbiology Paenibacillus Shuttle vector medicine Xylanase Escherichia coli Biotechnology
Zdroj:	Biocatalysis and Biotransformation. 25:157-162
ISSN:	1029-2446 1024-2422
DOI:	10.1080/10242420701379932
Popis:	Xylanase B from Paenibacillus barcinonensis was cloned in shuttle vectors for Escherichia coli and Bacillus subtilis, and expressed in Bacillus hosts. Several recombinant strains were constructed, among which B. subtilis MW15/pRBSPOX20 showed the highest production. This recombinant strain consists of a protease double mutant host containing P. barcinonensis xynB gene under the control of a phage SPO2 strong promoter. Maximum production was found when the strain was cultured in nutrient broth supplemented with xylans. Analysis of xylanase B location in B. subtilis MW15/pRBSPOX20 showed that the enzyme remained cell-associated in young cultures, consistent with its intracellular location in its original host, P. barcinonensis, and the lack of a signal peptide. However, when cultures reached the stationary phase, xylanase B was released to the external medium as a result of cell lysis. The amount of enzyme located in the supernatants of old cultures could account for 50% of total xylanase activity. Analysis...
Databáze:	OpenAIRE
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