GroEL-GroES solubilizes abundantly expressed xylulokinase in Escherichia coli.
OBJECTIVE
The aims of the present work were to solubilize the abundantly expressed recombinant xylulokinase in Escherichia coli and to develop a reliable xylulokinase assay.


RESULTS
Three mutants of xylulokinase of Bacillus megaterium that were expressed at high level but formed insoluble protein in E. coli BL21(DE3)pLysS were selected for solubility study. The solubility of xylulokinase increased eight to 77-fold after introduction of molecular chaperones GroEL-GroES into the host.


CONCLUSIONS
This investigation reports that GroEL-GroES minimizes the formation of insoluble protein in three highly expressed recombinant xylulokinases and an improved xylulokinase assay.


CONCLUSIONS
Commercial production of bioethanol is critically dependent on the development of an efficient and low-cost process of enzymatic conversion of xylan, a major component in lignocellulose biomass, to xylulose-5-phosphate, which can then be channelled into pentose phosphate pathway and metabolized to ethanol. The improved intracellular xylulokinase activity is expected to facilitate the xylose degradation.
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Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Tsang
P W K
PW
Fu
R Z
RZ
D019149:Bioreactors; D018835:Chaperonin 10 / Q000378:metabolism*; D018834:Chaperonin 60 / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D005285:Fermentation; D018832:Molecular Chaperones; D010428:Pentosephosphates / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000032:analysis / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014990:Xylans / Q000378:metabolism*; D014996:Xylulose / Q000378:metabolism
D018835:Chaperonin 10; D018834:Chaperonin 60; D018832:Molecular Chaperones; D010428:Pentosephosphates; D011994:Recombinant Proteins; D014990:Xylans; D014996:Xylulose; C031625:xylulose-5-phosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C032700:xylulokinase