Production and purification of a protease, a chitosanase, and chitin oligosaccharides by Bacillus cereus TKU022 fermentation.

A protease- and chitosanase-producing strain was isolated and identified as Bacillus cereus TKU022. The protease and chitosanase were both produced using 1.5% (w/v) shrimp head powder (SHP) as the sole carbon/nitrogen source, and these enzymes were purified from the culture supernatant. The molecular masses of the TKU022 protease and chitosanase determined using SDS-PAGE were approximately 45 and 44kDa, respectively. The high stability of the TKU022 protease toward surfactants, an optimal pH of 10 and an optimal temperature of 50-60°C suggest that this high-alkaline protease has potential applications for various industrial processes. Concomitant with the production of the TKU022 chitosanase, N-acetyl chitooligosaccharides were also observed in the culture supernatant, including (GlcNAc)(2), (GlcNAc)(4), (GlcNAc)(5), and (GlcNAc)(6) at concentrations of 201.5, 12.4, 0.5, and 0.3µg/mL, respectively, as determined using an HPLC analysis. The chitin oligosaccharides products were also characterized using a MALDI-TOF mass spectrometer. A combination of the HPLC and MALDI-TOF MS results showed that the chitin oligosaccharides of the TKU022 culture supernatant comprise oligomers with degree of polymerization (DP) from 2 to 6. Using this method, the production of a protease, a chitosanase, and chitin oligosaccharides may be useful for various industrial and biological applications.

Biodegradation and microbial community changes upon shrimp shell wastes amended in mangrove river sediment.

Chitin, a homopolymer of N-acetyl-D-glucosamine (GlcNAc) residues linked by beta 1-4 bonds, is the most abundant renewable natural resource after cellulose. It is widely distributed in nature as the integuments of crustaceans and insects and as a component of fungi and algae. This study investigated the effects of a bifunctional chitinase/lysozyme-producing strain, Pseudomonas aeruginosa K-187, on degradation of shrimp shells and the survival conditions of bacterial strains in mangrove river sediment of Tamsui River. The structures of the whole bacterial community of the samples were measured by using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique. Results show that three bacterial strains (Acrobacter sp., Shewanella sp., and Marinobacterium sp.) which originated from the mangrove river sediment were found predominant in the 6 days-incubation sample of P. aeruginosa K-187 amended mangrove river sediment. Meanwhile, biomass, reducing sugar, and total sugar were found highest in the 6 weeks-incubation sample of shrimp shell powder and P. aeruginosa K-187-amended mangrove river sediment. According to the results, we assumed that the amendment of P. aeruginosa K-187 can enhance the biodegradation of shrimp shells in the seawater containing mangrove river sediment. We hope that these findings may provide some useful information for the reclamation of chitin-containing wastes in our environment.

Conversion of shrimp shell by using Serratia sp. TKU017 fermentation for the production of enzymes and antioxidants.

A chitinase (CHT), and a protease (PRO) were purified from the culture supernatant of Serratia sp. TKU017 with shrimp shell as the sole carbon/nitrogen source. The molecular masses of CHT and PRO determined by SDS-PAGE were approximately 65 kDa and 53 kDa, respectively. CHT was inhibited by Mn2+, Cu2+ and PRO was inhibited by most tested divalent metals, EDTA. The optimum pH, optimum temperature, pH stability, and thermal stability of CHT and PRO were (pH 5, 50 degrees , pH 5 degrees ) and (pH 9, 40 degrees , pH 5 degrees ), respectively. PRO retained 95% of its protease activity in the presence of 0.5 mM SDS. The result demonstrates that PRO is SDS-resistant protease and probably has a rigid structure. The 4th day supernatant showed the strongest antioxidant activity (70%, DPPH scavenging ability) and the highest total phenolic content (196+/-6.2 microng of gallic acid equival/mL). Significant associations between the antioxidant potency and the total phenolic content, as well as between the antioxidant potency and free amino groups, were found for the supernatant. With this method, we have shown that shrimp shell wastes can be utilized and it's effective in the production of enzymes and antioxidants, facilitating its potential use in industrial applications and functional foods.