In comparison to bacterial alginate, algal alginate showed a minor binding capaticity. However,

binding of lipase to algal alginate was reported previously [34]. In contrast to bacterial alginate Enzalutamide mouse of P. aeruginosa, the algal alginate lacks O-acetyl groups and comprises a different monomer sequence which is characterized by the presence of guluronic acid rich regions (G-blocks) [22, 49]. Since other studies did not reveal an influence of the O-acetyl groups on the binding of lipases [33] the here observed effect might be based on the different monomer structure of algal and bacterial alginates. It was shown that https://www.selleckchem.com/products/amg510.html within the G-blocks of algal alginates specific intra- and intermolecular structures were formed (egg box). Within the egg boxes negative charges of the alginate molecules are directed to each other and are complexed via divalent cations.

Thereby, the negative charges were shielded [50]. Figure Anlotinib supplier 2 Binding of purified lipase LipA from P. aeruginosa to polysaccharides. Purified lipase LipA (36 ng/ml) from P. aeruginosa was incubated at 30°C in microtiter plates in the absence (−○-) and in the presence of immobilized polysaccharide films of (−■-) bacterial alginate from P. aeruginosa SG81 shown in red, (−▲-) xanthan shown in green, (−Δ-) algal alginate shown in pink, (−□-) levan shown in bright blue and (−●-) dextran shown in dark blue. Represented are carbohydrate concentrations used for coating of the microtiter plate wells. The bound lipase was Interleukin-2 receptor detected by activity measurements using pNPP as substrate. Results are shown as mean of five independent experiments +/− standard deviations. Significance of differences in lipase binding between coated and uncoated wells was calculated by ANOVA for the highest tested polysaccharide concentration. *** p < 0.001; ** p < 0.01. In summary, the experiments suggest that the

binding of lipases to alginate depends on the negative charged monomers of the polysaccharide indicating ionic interactions between the molecules. Heat stabilization of lipases by polysaccharides To investigate the biological impact of the interaction between lipase and bacterial alginate, heat inactivation experiments were performed. Incubation of purified lipase for 20 min at different temperatures in the presence and absence of polysaccharides showed an obvious influence of alginate on the stability of the lipase activity (Table 2). Without heat treatment (37°C) lipase activity stayed constant over 20 min in the presence and absence of polysaccharides at ΔA401 = 0.66 +/− 0.15 corresponding to an activity of 2.3 +/− 0.5 nmol/min × μg protein. Furthermore, at temperatures up to 50°C the lipase seemed to be generally stable. The addition of polysaccharides only had a minor effect. At higher temperatures (> 80°C) the protective effect of polysaccharides was lost. However, at approximately 70°C a significantly increased temperature tolerance of the lipase was observed in the presence of bacterial alginate.