Recombinant Expression of an Alkali Stable GH10 Xylanase from Paenibacillus barcinonensis

• Published in: Journal of agricultural and food chemistry Vol. 58; no. 8; pp. 4814 - 4818
• Main Authors: Valenzuela, Susana V, Díaz, Pilar, Javier Pastor, F. I
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 28.04.2010
• Subjects: alkali stable GH10 xylanase; Alkalies; amino acid sequences; Base Sequence; Biological and medical sciences; chemical analysis; Chemical Aspects of Biotechnology/Molecular Biology; chemical composition; Cloning, Molecular; DNA Primers; Endo-1,4-beta Xylanases - genetics; Endo-1,4-beta Xylanases - metabolism; enzyme activity; Food industries; Fundamental and applied biological sciences. Psychology; molecular sequence data; new species; novel foods; Paenibacillus; Paenibacillus - enzymology; Paenibacillus - genetics; Paenibacillus barcinonensis; protein synthesis; pulp and paper industry; recombinant proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; xylanases; Biotechnology; Paenibacillus; alkali stable; enzymatic hydrolysis; bioconversion; xylanase; Enzyme; Glycosylases; Hydrolysis; Alkali; Glycosidases; Hydrolases; Xylan endo-1,3-β-xylosidase; Enzymatic reaction
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf9045792

Xylanase A from Paenibacillus barcinonensis, a new species isolated from a rice field, has been cloned and expressed in Escherichia coli. Purified recombinant xylanase showed high activity on xylans from hardwoods and cereals, and exhibited K m and V max of 2.93 mg/mL and 50.67 U/mg on birchwood xylan. Xylanase A was highly active at 60 °C in alkaline pH values up to 9.5 and remained stable for at least 3 h in alkaline conditions. The amino acid sequence deduced from xynA revealed that it is a single domain xylanase belonging to the GH10 family. Thin layer chromatography analysis showed that the enzyme released a mixture of hydrolysis products including substituted xylooligomers from cereal arabinoxylans, while xylose, xylobiose, and aldotetraouronic acid were the main products released from glucuronoxylan from birchwood. The enzyme released a complex mixture of xylooligomers for acetylated xylan from eucalyptus, revealing its potential to depolymerize this widely used resource in the pulp and paper industry.