Production of 5′-phosphodiesterase by Catharanthus roseus cells promoted by heat-degraded products generated from uronic acid

Polyalginate was autoclaved at 121degreesC for 20 min and its molecular weight distribution was analyzed. The autoclaved alginate yielded alginate polymer, oligomer and heat degraded products (HDPs). Each of the separated substances promoted 5'-phosphodiesterase (5'-PDase) production in suspension culture of Catharanthus roseus cells. HDPs could also be generated from other uronic acids (galacturonic acid and glucuronic acid) by autoclave treatment. The most effective substance in the HDPs was isolated and characterized as trans-4,5-dihydroxy-2-cyclopenten-1-one (DHCP). The optimal conditions for DHCP production were also established (autoclaving 1 mg/ml monogalacturonic acid [pH 2] at 121degreesC for 2 h). A combination of oligo-alginate (below 4 kDa) and HDPs significantly promoted the production of 5'-PDase in C. roseus. Based on the above results, a novel alginate complex that gave a 44-fold increase in 5'-PDase production by C. roseus was developed.