Cloning and overexpression of rhamnose isomerase and fucose isomerase.
Rhamnose isomerase and fucose isomerase were overexpressed in E. coli, purified and characterized. The rhamnose isomerase gene was ligated to the restriction sites of PstI and Hind III of vector pTrcHis and the fucose isomerase gene was ligated to the EcoRI and PstI sites of vector pKK223-3 for overexpression of the enzymes in E. coli XL1-Blue MRF. Approximately 16,500 U of active fucose isomerase and 2400 of rhamnose isomerase can be obtained per liter of culture from these expression systems.
Garcia-Junceda
E
E
Department of Chemistry, Scripps Research Institute, La Jolla, CA 92037, USA.
Shen
G J
GJ
Alajarin
R
R
Wong
C H
CH
D019747:Aldose-Ketose Isomerases; D000818:Animals; D001483:Base Sequence; D002238:Carbohydrate Epimerases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins; D002238:Carbohydrate Epimerases; D019747:Aldose-Ketose Isomerases; C062878:L-rhamnose isomerase; C019714:D-arabinose isomerase