University of Illinois Urbana-Champaign

Over-expression and characterization of four alginate lyases from vibrio splendidus 12B01

Badur, Ahmet

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https://hdl.handle.net/2142/73046

Description

Title
Over-expression and characterization of four alginate lyases from vibrio splendidus 12B01
Author(s)
Badur, Ahmet
Issue Date
2015-01-21
Director of Research (if dissertation) or Advisor (if thesis)
Rao, Christopher V.
Department of Study
Chemical & Biomolecular Engr
Discipline
Chemical Engineering
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
M.S.
Degree Level
Thesis
Keyword(s)
  • Vibrio splendidus
  • Vibrio splendidus 12B01
  • alginate
  • alginate lyase
  • poly-guluronate (polyG)
  • poly-mannuronate (polyM)
  • heteropolymeric M/G (polyMG)
  • polyGM
  • AlyA
  • AlyB
  • AlyC
  • AlyD
  • AlyE
Abstract
lginate is a polysaccharide found within brown seaweeds and has been targeted as a carbon source for biofuel production. Alginate consists of α-L-guluronate (G) and β-D-mannuronate (M) linked in various patterns, which results in either a homo- or heteropolymeric structure. Alginate lyases are enzymes that degrade the linkage between G and M blocks and can have specificity to either polyG, polyM, or polyMG block degradation. The marine bacterium Vibrio splendidus 12B01 contains four putative alginate lyases which were investigated in this study. We identified, purified, and characterized the four PL7 alginates lyases. We found that these lyases have optimal activity between pH 7.5-8.5 and 20-25°C, consistent with use in a marine environment. Additionally, Ca 2+ is necessary for optimal enzyme activity. The binding constant (K m ) of the lyases toward alginate was found to be between 22 and 123 mM alginate and the maximum reaction rate (V max ) was found to be between 0.13 and 0.83 μM s -1 . The turnover numbers for the lyases was found to be between 0.60 and 7.1 s -1 .
Graduation Semester
2014-12
Permalink
http://hdl.handle.net/2142/73046
Copyright and License Information
Copyright 2014 Ahmet Badur

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