Thermogravimetric evaluation of chitin degradation in soil: implication for the enhancement of ammonification of native organic nitrogen by chitin addition

Degradation of chitin, which is an aminopolysaccharide used as a soil amendment, has been often monitored in soil via its degradation products such as carbon dioxide and ammonium. We report here the applicability of thermogravimetry to measure the amount of chitin added to soil. The maximum pyrolysis rate of the upland surface soil of Brown Forest soil supplemented with chitin was strongly correlated with added chitin content (r = 0.999) when the content exceeded 6.0 g kg^?1. The maximum pyrolysis rates of chitin-added soil (around 385°C) was distinctive from those of soil supplemented with cellulose, chitosan, N-acetylglucosamine, and N,N’-diacetylchitobiose (around 340°C, 300°C, 200°C, and 240°C, respectively), indicating the specific detection of chitin. Soil incubation study demonstrated that 60 g kg^?1 chitin added to the soil declined exponentially (r = 0.993) within days and could not be detected at 90 days after the addition of chitin. Total carbon (C) content also decreased within days whereas total nitrogen (N) remained almost constant over the 90 days. The amount of ammonium-N increased in the initial 30 days after the addition of chitin and reached about 3.6 g kg^?1, which corresponded to the amount of N in the added chitin (4.1 g kg^?1) while the amount of nitrite-N and nitrate-N were below 2.0 and 15 mg kg^?1, respectively. Comparison of the measured ammonium-N and total-C contents with those calculated from the measured chitin-content implied that addition of chitin enhanced degradation of native organic compounds in soil.     

Identification and Characterization of Three Chitinases with Potential in Direct Conversion of Crystalline Chitin into N,N′-diacetylchitobiose

Chitooligosaccharides (COSs) have been widely used in agriculture, medicine, cosmetics, and foods, which are commonly prepared from chitin with chitinases. So far, while most COSs are prepared from colloidal chitin, chitinases used in preparing COSs directly from natural crystalline chitin are less reported. Here, we characterize three chitinases, which were identified from the marine bacterium Pseudoalteromonas flavipulchra DSM 14401^T, with an ability to degrade crystalline chitin into (GlcNAc)₂ (N,N’-diacetylchitobiose). Strain DSM 14401 can degrade the crystalline α-chitin in the medium to provide nutrients for growth. Genome and secretome analyses indicate that this strain secretes six chitinolytic enzymes, among which chitinases Chia4287, Chib0431, and Chib0434 have higher abundance than the others, suggesting their importance in crystalline α-chitin degradation. These three chitinases were heterologously expressed, purified, and characterized. They are all active on crystalline α-chitin, with temperature optima of 45–50 °C and pH optima of 7.0–7.5. They are all stable at 40 °C and in the pH range of 5.0–11.0. Moreover, they all have excellent salt tolerance, retaining more than 92% activity after incubation in 5 M NaCl for 10 h at 4 °C. When acting on crystalline α-chitin, the main products of the three chitinases are all (GlcNAc)₂, which suggests that chitinases Chia4287, Chib0431, and Chib0434 likely have potential in direct conversion of crystalline chitin into (GlcNAc)₂.