Heterologous expression of an alginate lyase from Streptomyces sp. ALG-5 in Escherichia coli and its use for preparation of the magnetic nanoparticle-immobilized enzymes.
The marine alginate lyase from Streptomyces sp. ALG-5, which specifically degrades poly-G block of alginate, was functionally expressed as a His-tagged form with an Escherichia coli expression system. The recombinant alginate lyase expressed with pColdI at 15 °C exhibited the highest alginate-degrading activity. The recombinant alginate lyase was efficiently immobilized onto two types of magnetic nanoparticles, superparamagnetic iron oxide nanoparticle, and hybrid magnetic silica nanoparticle, based on the affinity between His-tag and Ni(2+) that displayed on the surfaces of nanoparticles. An alginate oligosaccharide mixture consisting of dimer and trimer was prepared by the immobilized alginate lyase. The immobilized enzymes were re-used repeatedly more than 10 times after magnetic separation.
Shin
Jung Won
JW
Department of Chemical Engineering, Industrial Liaison Research Center, Kyung Hee University, Gyeonggi-do 446-701, Republic of Korea.
Choi
Sung Hee
SH
Kim
Dong Eun
DE
Kim
Hee Sook
HS
Lee
Jae-Hwa
JH
Lee
In Su
IS
Lee
Eun Yeol
EY
D000464:Alginates / Q000378:metabolism; D003001:Cloning, Molecular; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008280:Magnetics; D053758:Nanoparticles / Q000737:chemistry; D011133:Polysaccharide-Lyases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D013302:Streptomyces / Q000201:enzymology*
D000464:Alginates; D004800:Enzymes, Immobilized; D011993:Recombinant Fusion Proteins; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase