Characterization of a novel cellulose biosynthesis inhibitor, CBI28, in Gluconacetobacter xylinus

To study the underlying mechanisms for microbial cellulose biosythesis, a novel compound, CBI28, was used as an inhibitor along with classical genetics and EMS mutagenesis. An EZ-Link Biotin Hydrazide Kit was used to create a CBI28-Biotin conjugate for further studies. Gluconacetobacter xylinus cells were exposed to 10 uM CBI28 to induce cellulose biosythesis inhibition, lysed and small hydrophobic molecules were extracted using methanol and Waters Oasis HLB SPE-Paks. Samples were separated and detected using the Ultra Performance Liquid Chromatograph-Mass Spectrometer/Photo Diode Array. Putative mutants were isolated but did not survive for further study. An ion with the expected mass of a CBI28-Biotin conjugate (552 m/z) was detected but not in sufficiently high concentrations for characterization. Metabolite studies revealed putative metabolites derived from the HLB SPE and methanol extractions with no significant difference in extraction methods. Potential metabolites with masses of ~281.77 m/z and ~79 m/z were detected in CBI28 exposed cells. Further analysis needs to be performed to determine if CBI28 metabolites prevent cellulose production.

Name thesis:degree_name

Master of Science (MSc)

Discipline thesis:degree_discipline

Applied Bioscience

Grantor

University of Ontario Institute of Technology

Year dc:date.issued

2010

Author dc:creator

• Harripaul, Ricardo Simeon

Advisor dc:contributor.advisor

• Forbes, Shari L.

• Cellulose
• Cellulose biosynthesis inhibitor
• CBI28
• Gluconacetobacter xylinus

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10155/122

OAI identifier oai:identifier

oai:ontariotechu.scholaris.ca:10155/122