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

Badur, Ahmet

Permalink

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

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

Owning Collections