Strategies for the anisodamine production by a recombinant Saccharomyces cerevisiae strain

CARDILLO, ALEJANDRA BEATRIZ; QUEVEDO, CARLA; PERASSOLO, MARÍA; MARTINEZ, CAROLINA; BUSTO, VICTOR; RODRIGUEZ TALOU, JULIAN; GIULIETTI, ANA MARIA

Villa Carlos Paz

Congreso; SAIB 2008; 2008

Brugmansia candida is a South American native plant that produces anisodamine, scopolamine, and hyosciamine. The last ones are widely used as pharmaceuticals. Recently, alternative medical applications for anisodamine were described. The medical use of this alkaloid has many advantages since it appears to be less toxic than atropine and it has less negative effects over CNS than scopolamine. Hyoscyamine 6ß-hydroxylase (H6H, EC 1.14.11.11) catalyzes the conversion of hyoscyamine into anisodamine and scopolamine. The use of an immobilized system in biotransformation processes has many advantages comparing to the use of free cells. The aim of this work was to analyze the ability of the recombinant Saccharomyces cerevisiae strain carrying the h6hDNAc to produce anisodamine by the free and immobilized cell system. Also, the free and immobilized crude protein extract from this strain was evaluated. Alginate gel 1% was used for the entrapment. The effect of the immobilization process and different treatments of cell permeabilization were also compared. According to the results obtained in this work, the immobilization process was disadvantageous. The conversion of hyoscyamine by the free protein extract was more efficient (reaching an 83% of anisodamine) than the conversion of the alkaloid by the immobilized protein extract and the free and immobilized transformed S. cerevisiae cells.