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6346916 
Journal Article 
Efficient solubilization, purification of recombinant extracellular [alpha]-amylase from pyrococcus furiosus expressed as inclusion bodies in Escherichia coli 
Wang, L; Zhou, Q; Chen, H; Chu, Z; Lu, J; Zhang, Y; Yang, S 
2007 
Yes 
Journal of Industrial Microbiology and Biotechnology
ISSN: 1367-5435
EISSN: 1476-5535 
Journal of Industrial Microbiology & Biotechnology 
34 
3 
187-187 
English 
17119903 
WOS:000244066500002 
The gene encoding the Pyrococcus furiosus extracellular alpha-amylase (PFA) was amplified by PCR from P. furiosus genomic DNA and was highly expressed in Escherichia coli BL21-Codon Plus (DE3)-RIL. The recombinant alpha-amylase was mainly expressed in the form of insoluble inclusion bodies. An improved purification method was established in this paper. The solubilization of the inclusion bodies was achieved by 90 degrees C treatment for 3 min in Britton-Robinson buffer at pH 10.5. The solubilized PFA was then diluted and subsequently purified by Phenyl Sepharose chromatography. The overall yield of the new purification method was about 58,000 U/g wet cells, which is higher than previously reported. 
Enzyme Stability; Escherichia coli/genetics; Escherichia coli/ultrastructure; Hydrogen-Ion Concentration; Inclusion Bodies/enzymology; Pyrococcus furiosus/enzymology; Recombinant Proteins/isolation & purification; Temperature; alpha-Amylases/genetics; alpha-Amylases/isolation & purification; alpha-Amylases/metabolism; Microbiology/