Biochemical characterization of a novel bifunctional chitosanase from Paenibacillus barengoltzii for chitooligosaccharide production.

A novel chitosanase gene, designated as PbCsn8, was cloned from Paenibacillus barengoltzii. It shared the highest identity of 73% with the glycoside hydrolase (GH) family 8 chitosanase from Bacillus thuringiensis JAM-GG01. The gene was heterologously expressed in Bacillus subtilis as an extracellular protein, and the highest chitosanase yield of 1, 108 U/mL was obtained by high-cell density fermentation in a 5-L fermentor. The recombinant chitosanase (PbCsn8) was purified to homogeneity and biochemically characterized. PbCsn8 was most active at pH 5.5 and 70 °C, respectively. It was stable in a wide pH range of 5.0-11.0 and up to 55 °C. PbCsn8 was a bifunctional enzyme, exhibiting both chitosanase and glucanase activities, with the highest specificity towards chitosan (360 U/mg), followed by barley ß-glucan (72 U/mg) and lichenan (13 U/mg). It hydrolyzed chitosan to release mainly chitooligosaccharides (COSs) with degree of polymerization (DP) 2-3, while hydrolyzed barley ß-glucan to yield mainly glucooligosaccharides with DP > 5. PbCsn8 was further applied in COS production, and the highest COS yield of 79.3% (w/w) was obtained. This is the first report on a GH family 8 chitosanase from P. barengoltzii. The high yield and remarkable hydrolysis properties may make PbCsn8 a good candidate in industrial application.

An experimental comparison: Horizontal evaluation of valuable metal extraction and arsenic emission characteristics of tailings from different copper smelting slag recovery processes.

This study comprehensively investigated arsenic's enrichment, distribution, and characteristics in tailings. XRD and SEM-EDS characterized the phase and morphology of tailings from various smelting processes. At the same time, the embedding characteristics of arsenic in the ore phase were analyzed by EPMA. The differences between arsenic's leading ore phase carriers in different recovery processes were found. It was discussed that this phenomenon would be related to the element-binding ability and the precipitation priority of the ore phase. The occurrence state of arsenic was discussed by sequential chemical extraction experiments. The proportion of leachable arsenic is higher than the low-risk limit, whatever which smelting method is adopted, which leads to high environmental risk. In the experiment of comparing the leaching toxicity of tailings by different leaching methods, the arsenic concentration in the leaching solution of tailings recovered by the flotation method exceeds the specified safety range. Although the tailings after reduction smelting did not show high leaching toxicity, a large number of accumulations also would not represent absolute safety.