Isolating thermostable enzymes from engineered mesophiles

Details Isolating thermostable enzymes from engineered mesophiles Isolating thermostable enzymes from engineered mesophiles

C - Chemistry, Metallurgy

12

N

150/15, 195/1.23

C12N 15/52 (2006.01) C12N 9/00 (2006.01) C12Q 1/25 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2005667

Abstract of the Disclosure Isolating a thermostable enzyme free from unwanted contaminants by: (a) providing a mesophilic host cell engineered to express a gene encoding a heterologous thermostable enzyme: (b) culturing the mesophilic host cell and producing the thermostable enzyme in a mixture comprising unwanted contaminants; and (c) purifying the thermostable enzyme by a method that includes heating the enzyme and the unwanted contaminants to a temperature sufficient to inactivate the unwanted contaminants but not sufficient to inactivate the thermostable enzyme. A method of assaying for nucleic acid ligase includes forming an adenyl adduct with the ligase and detecting the adduct. The latter method is used to screen a library of transformants for the ability to express the nucleic acid ligase.

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