Crystallization of enzyme and method for producing same

Abstract

The present invention provides a method of crystallizing of enzymes. The method is for rapidly crystallizing enzymes from impure mixtures. The method is simple and cheap, and it is compatible to industrial requirements. T1 lipase was able to form crystals at low protein concentration (2.5 mg/ml) in a day. High temperature crystallization was obtained from the method. The present invention also relates to a composition of a crystallized lipase produced from the said method.

1 . A purified T1 lipase enzyme crystal obtained from a method comprising
 a) providing purified T1 lipase to be crystallized,   b) preparing a crystallization mix comprising
 a buffer comprising 0.1 M NaH 2 PO 4 , 0.1 M K H 2 PO 4 , 2.0 M NaCl and 0.1 M MES pH 6.5, 
 between 0.5 to 4.5 mg/mL of lipase from step (a), and 
 between 1 an 2M of precipitant, and 
   c) obtaining T1 lipase crystals from step (b) by a hanging-drop methodology, wherein crystals of the lipase are formed between about 1 hour and 1 day, and   wherein said enzyme is produced from  Geobacillus  sp. strain T1.   
   
   
       2 . The enzyme crystal as claimed in  claim 1 , wherein said enzyme is a purified T1 lipase. 
   
   
       3 . The enzyme crystal as claimed in  claim 1 , wherein T1 lipase is T1 mature lipase and/or T1 fusion lipase. 
   
   
       4 . A composition comprising a crystalline lipase from  Geobacillus  sp. Strain T1, produced by a method comprising
 a) providing purified T1 lipase to be crystallized,   b) preparing a crystallization mix comprising
 a buffer comprising 0.1 M NaH 2 PO 4 , 0.1 M K H 2 PO 4 , 2.0 M NaCl and 0.1 M MES pH 6.5, 
 between 0.5 to 4.5 mg/mL of lipase from step (a), and 
 between 1 an 2M of precipitant, and 
   c) obtaining T1 lipase crystals from step (b) by a hanging-drop methodology, wherein crystals of the lipase are formed between about 1 hour and 1 day,