Molecular cloning of levan fructotransferase gene from Arthrobacter ureafaciens K2032 and its expression in Escherichia coli for the production of difructose dianhydride IV.
OBJECTIVE
To clone and overexpress a novel levan fructotransferase gene lftA from Arthrobacter ureafaciens K2032.


RESULTS
The lftA gene, encoding a levan fructotransferase (LFTase) of 521 amino acids (aa) residues, was cloned from the genomic DNA of A. ureafaciens K2032, and overexpressed in Escherichia coli. The recombinant LFTase overexpressed in E. coli was then used to produce a difructose dianhydride (DFA IV) from levan. DFA IV crystals with 97% purity could be obtained from the reaction mixture in 83.7% yield by using a natural crystallization method.


CONCLUSIONS
The lftA gene cloned from A. ureafaciens K2032 encode a novel levan fructotransferase which produces difructose dianhydride (DFA IV) from levan.


CONCLUSIONS
Levan fructotransferase is a useful enzyme with great promise in the production of DFA IV and various fructosides.
Kim
C H
CH
Laboratory of Metabolic Engineering, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea. kim3641@kribb.re.kr
Jang
E K
EK
Kim
S H
SH
Jang
K H
KH
Kang
S A
SA
Song
K B
KB
Kwon
O S
OS
Rhee
S K
SK
D000595:Amino Acid Sequence; D001173:Arthrobacter / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D004187:Disaccharides / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics*; D006602:Hexosyltransferases / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics
D004187:Disaccharides; D011994:Recombinant Proteins; C033869:difructose anhydride IV; D006602:Hexosyltransferases; C040074:levan fructotransferase