Development of novel alginate lyase cross-linked aggregates for the oral treatment of cystic fibrosis

ISLAN GA; MARTINEZ YN; ILLANES ANDRES; CASTRO GR

RSC Advances

Royal Society of Chemistry,

Lugar: Cambridge; Año: 2014 vol. 4 p. 11758 - 11765

2046-2069

Alginate lyase (AL) from Sphingobacterium multivorum is unsuitable for oral delivery because of its rapidinactivation under acidic conditions. The synthesis of a novel crosslinking enzyme aggregate (CLEA) ofAL (AL-CLEA) is proposed. AL precipitation with 95% ammonium sulfate and combined with lowmethoxylated pectin (LMP), showed 100% precipitation yield. Crosslinking with glutaraldehyde reducedthe AL-CLEA activity to less than 1%, but addition of bovine serum albumin (BSA) and LMP duringAL-CLEA synthesis increased the activity yield to 14.7%. AL-CLEA exposed to simulated gastric conditions(pH 1.2 to 3.0) showed more than 70% retention of enzymatic activity. Moreover, AL-CLEA showedthermal stability at temperatures over 37
C. Stability against chemical denaturants (ethanol, acetone andpropylene glycol) showed that AL-CLEA was 14 times more stable than free AL in all cases. Finally, a 25%viscosity reduction of alginate solution was achieved with AL-CLEA. This is the first report of AL-CLEAsynthesis and evaluation.