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

Vibrio splendidus;Vibrio splendidus 12B01;alginate;alginate lyase;poly-guluronate (polyG);poly-mannuronate (polyM);heteropolymeric M/G (polyMG);polyGM;AlyA;AlyB;AlyC;AlyD;AlyE

Badur, Ahmet ; Rao ; Christopher V.

关键词: Vibrio splendidus;    Vibrio splendidus 12B01;    alginate;    alginate lyase;    poly-guluronate (polyG);    poly-mannuronate (polyM);    heteropolymeric M/G (polyMG);    polyGM;    AlyA;    AlyB;    AlyC;    AlyD;    AlyE;   

Others  :  https://www.ideals.illinois.edu/bitstream/handle/2142/73046/Ahmet_Badur.pdf?sequence=1&isAllowed=y

美国|英语

来源: The Illinois Digital Environment for Access to Learning and Scholarship

【 摘 要 】

lginate is a polysaccharide found within brown seaweeds and has beentargeted 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 linkagebetween G and M blocks and can have specificity to either polyG, polyM, or polyMGblock degradation. The marine bacterium Vibrio splendidus 12B01 contains fourputative alginate lyases which were investigated in this study. We identified, purified,and characterized the four PL7 alginates lyases. We found that these lyases haveoptimal activity between pH 7.5-8.5 and 20-25°C, consistent with use in a marineenvironment. Additionally, Ca 2+ is necessary for optimal enzyme activity. The bindingconstant (K m ) of the lyases toward alginate was found to be between 22 and 123 mMalginate and the maximum reaction rate (V max ) was found to be between 0.13 and 0.83μM s -1 . The turnover numbers for the lyases was found to be between 0.60 and 7.1 s -1 .