Construction, expression and characterization of fusion enzyme from Arthrobacter oxydans dextranase and Klebsiella pneumoniae amylase.
An artificial fusion protein of Arthrobacter oxydans dextranase and Klebsiella pneumoniae alpha-amylase was constructed and expressed in Escherichia coli. Most of the expressed protein existed as an insoluble fraction, which was solubilized with urea. The purified fusion enzyme electrophoretically migrated as a single protein band; M = 137 kDa, and exhibited activities of both dextranase (10.8 U mg(-1)) and amylase (7.1 U mg(-1)), which were lower than that of reference dextranase (13.3 U mg(-1)) and alpha-amylase (103 U mg(-1)). The fusion enzyme displayed bifunctional enzyme activity at pH 5-7 at 37 degrees C. These attributes potentially make the fusion enzyme more convenient for use in sugar processing than a two-enzyme system.
Kim
Young-Min
YM
School of Biological Sciences and Technology, The Research Institute for Catalysis, Chonnam National University, Gwangju, 500-757, Korea.
Ko
Eun-Ah
EA
Kang
Hee-Kyoung
HK
Kim
Doman
D
D000681:Amylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001173:Arthrobacter / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003910:Dextranase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007711:Klebsiella pneumoniae / Q000201:enzymology* / Q000235:genetics; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D000681:Amylases; D003910:Dextranase