Isolation, expression, purification and characterisation of a novel acetyl xylan esterase from streptomyces species ORS10

Simple item page
dc.contributor.authorGao, Yu
dc.contributor.otherDept. of Biotechnology
dc.contributor.otherFaculty of Science
dc.date.accessioned2014-02-10T14:10:39Z
dc.date.accessioned2024-05-09T07:44:59Z
dc.date.available2012/06/15 12:28
dc.date.available2012/06/15
dc.date.available2014-02-10T14:10:39Z
dc.date.available2024-05-09T07:44:59Z
dc.date.issued2012
dc.descriptionMagister Scientiae - MScen_US
dc.description.abstractLignocellulosic biomass represents an important renewable resource for biofuels production. Lignocellulosic biomass is comprised of cellulose, hemicellulose and lignin. Lignocellulosics are highly recalcitrant to enzymatic degradation and due to its complex nature a range of enzymes are required to synergistically hydrolyse biomass. Many microorganisms are capable of producing these enzymes as part of their hemicellulolytic hydrolysis system(s). The aim of this study was the characterisation of a thermophilic actinobacterial isolate (ORS10), capable of producing hemicellulosic enzymes, and the cloning and characterization of a hemicellulosic enzyme produced by the isolate. Phylogenetic analyses clustered ORS10 with species of the genus Streptomyces. BLAST analysis revealed that ORS10 was most closely related to Streptomyces achromogenes (99% identity). A small-insert genomic library was constructed and a putative acetylxylan esterase (AXEase) gene, axe10, was identified. The enzyme, Axe10, has moderate similarity to α/β hydrolase proteins, and contains an esterase/lipase superfamily conserved domain and a typical AXEase catalytic triad. The axe10 gene was sub-cloned into an expression vector [pET21a(+)] and a 28.7 kDa protein with demonstrated AXE activity was purified from E. coli Rosetta (DE3) pLysS. Axe10 displayed optimum activity at 37oC and pH 7.0. Despite being derived from a thermophilic Streptomyces species Axe10 was not thermostable. However, given the relative novelty of Axe10, further characterisation and assessment of this enzyme is warranted.en_US
dc.description.countrySouth Africa
dc.identifier.urihttps://hdl.handle.net/10566/13258
dc.language.isoenen_US
dc.publisherUniversity of the Western Capeen_US
dc.rights.holderUniversity of the Western Capeen_US
dc.subjectLignocellulosic biomassen_US
dc.subjectmicroorganismsen_US
dc.subjecthydrolyse biomassen_US
dc.subjectenzymesen_US
dc.subjectstreptomycesen_US
dc.subjectesteraseen_US
dc.subjectstreptomyces achromogenesen_US
dc.subjectphylogenetic analysesen_US
dc.titleIsolation, expression, purification and characterisation of a novel acetyl xylan esterase from streptomyces species ORS10en_US
dc.typeThesisen_US

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Gao_MSC_2012.pdf
Size:
2.63 MB
Format:
Adobe Portable Document Format
Download

Collections

Magister Scientiae - MSc (Biotechnology)