Enhancement of chitosanase production by cell immobilization of Gongronella sp. JG

Chitosanase production of Gongronella sp. JG cells immobilized in calcium alginate gel and polyurethane foam was compared with that of the free cells, there was a 60% increase in the enzyme yield (2429 U/L) compared to the highest yield obtained from free cells (1513 U/L). The optimal immobilization parameters (concentrations of sodium alginate, calcium chloride, bead inoculums, bead diameter, etc) for the enhanced production of chitosanase were determined as: sodium alginate 2% (w/v), 0.1 M calcium chloride, inoculum 10 mL beads to 100 mL production media and 2.7 mm bead diameter. Maximum chitosanase production was achieved with initial pH of 5.5 and temperature of 30 ºC. The alginate beads had well stability, retained 85% ability of enzyme production even after 7 cycles of repeated batch fermentation. These results showed the immobilization technique was a feasible and economical method for chitosansase production by Gongronella sp. JG.

Chitosanase production by Paenibacillus ehimensis and its application for chitosan hydrolysis

The chitosanase production by Paenibacillus ehimensis was studied in submerged cultures and the chitosan hydrolysis was evaluated by using these enzymes without purification. The bacterium produced inducibles enzymes after 12 h of growth in a culture medium containing 0.2 percent (w/v) of soluble chitosan as carbon source. The enzyme production was strongly repressed by the presence of glucose. The production started as soon as the available sugars finished in the culture medium. The maximum level of chitosanase activity was 500 U.L-1 at 36°C after 36 h incubation. The crude enzyme was optimally active at pH 6.0 and 55°C and in these conditions, the enzyme presented good stability (6 days). The enzyme without purification was used to hydrolyze the chitosan which resulted chitooligosaccharides between 20 and 30 min of reaction.

A produção de quitosanases pelo Paenibacillus ehimensis foi estudada em culturas submersas e a hidrólise da quitosana foi realizada utilizando essas enzimas sem purificação. As enzimas foram obtidas após 12 horas de crescimento desta bactéria em meio de cultivo contendo 0,2 por cento (p/v) de quitosana solúvel como fonte de carbono. A produção das enzimas foi fortemente reprimida na presença de glicose, sendo obtida após o consumo total dos açúcares disponibilizados no referido meio de cultivo. A máxima atividade quitosanolítica foi obtida após 36 horas de cultivo a 36ºC, atingindo valores de 500 U.L-1. As enzimas utilizadas no extrato bruto apresentaram melhores atividades quando submetidas a condições de pH e temperatura de 6,0 e 55ºC, respectivamente, e nessas condições permaneceram estáveis durante 6 dias. Essas enzimas sem serem submetidas aos processos de purificação foram utilizadas para hidrolisar a quitosana, obtendo os quito-oligossacarídeos entre 20 e 30 minutos de reação.