Method for stabilizing nitrilase activity and preserving...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/78 (2006.01) C12N 1/04 (2006.01) C12N 9/96 (2006.01) C12N 11/04 (2006.01) C12N 11/08 (2006.01) C12N 11/10 (2006.01) C12P 13/00 (2006.01)

Patent

CA 2377101

A method for preserving immobilized or unimmobilized microbial cells having nitrilase activity and for stabilizing the nitrilase activity of unimmobilized or immobilized microbial cells has been developed. The unimmobilized or immobilized microbial cells are stored in an aqueous solution containing from about 0.10 M to the saturation concentration of an inorganic salt of bicarbonate or carbonate, including ammonium, sodium and potassium salts of bicarbonate or carbonate. Aqueous suspensions containing at least 100 mM bicarbonate or carbonate limit microbial contamination of the stored enzyme catalyst, as well as stabilize the desired nitrilase activity of the unimmobilized or immobilized cells. Microorganisms which are characterized by a nitrilase activity and are stabilized and preserved by this method include <i>Acidovorax facilis</i> 72-PF-15 (ATCC 55747), <i>Acidovorax facilis</i> 72- PF-17 (ATCC 55745), <i>Acidovorax facilis</i> 72W (ATCC 55746), and transformed microbial cells having nitrilase activity, preferably <i>E. coli</i> SS1001 (ATTCC PTA-1177) which is transformed with Acidovorax facilis 72W nitrilase activity.

L'invention se rapporte à un procédé permettant de conserver des cellules microbiennes immobilisées ou non immobilisées et dotées d'une activité de nitrilase et de stabiliser l'activité de nitrilase de cellules microbiennes immobilisées ou non immobilisées. Ces cellules microbiennes immobilisées ou non immobilisées sont conservées dans une solution aqueuse contenant une quantité comprise entre 0,10 M environ et la concentration de saturation d'un bicarbonate ou carbonate inorganique, et notamment des bicarbonates ou carbonates d'ammonium, de sodium et de potassium. L'invention se rapporte également à des suspensions aqueuses contenant au moins 100 mM de bicarbonate ou de carbonate, qui limitent la contamination microbienne du catalyseur enzymatique stocké, et stabilisent également l'activité de nitrilase souhaitée des cellules non immobilisées ou immobilisées. Les micro-organismes qui sont caractérisés par une activité de nitrilase et sont stabilisés et conservés par ce procédé incluent <i>Acidovorax facilis</i> 72-PF-15 (ATCC 55747), <i>Acidovorax facilis</i> 72-PF-17 (ATCC 55745), <i>Acidovorax facilis</i> 72W (ATCC 55746), et les cellules microbiennes transformées dotées d'une activité de nitrilase sont de préférence <i>E. coli</i> SS1001 (ATTCC PTA-1177) qui est transformée de manière à être dotée de l'activité de nitrilase de <i>Acidovorax facilis</i> 72W.

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