N-terminal fusion of a hyperthermophilic chitin-binding domain to xylose isomerase from Thermotoga neapolitana enhances kinetics and thermostability of both free and immobilized enzymes.

ABSTRACT: Immobilization of a thermostable D-xylose isomerase (EC 5.3.1.5) from Thermotoga neapolitana 5068 (TNXI) on chitin beads was accomplished via a N-terminal fusion with a chitin-binding domain (CBD) from a hyperthermophilic chitinase produced by Pyrococcus furiosus (PF1233) to create a fusion protein (CBD-TNXI). The turnover numbers for glucose to fructose conversion for both unbound and immobilized CBD-TNXI were greater than the wild-type enzyme: k(cat) (min(-1)) was approximately 1,000, 3,800, and 5,800 at 80 degrees C compared to 1,140, 10,350, and 7,000 at 90 degrees C, for the wild-type, unbound, and immobilized enzymes, respectively. These k(cat) values for the glucose to fructose isomerization measured are the highest reported to date for any XI at any temperature. Enzyme kinetic inac

SUBMITTER: Harris JM 

PROVIDER: S-EPMC3711014 | biostudies-literature | 2010 Jul-Aug

REPOSITORIES: biostudies-literature