壳寡糖的制备、分离分析方法及在农业上的应用

壳寡糖具有独特的生理活性和功能性质,在多个领域有广泛的用途,尤其是在农业中作为植物病原菌生长抑制剂、土壤修复剂、产生诱导抗性等,逐渐成为国内外关注热点。其主要的制备方法有化学法、酶解法和物理法。分离方法主要有色谱柱分离法、膜分离法、酶法。分析方法有高效液相色谱法、质谱分析、核磁共振和红外光谱。     

环保型缓蚀剂壳寡糖对304不锈钢化学机械抛光的影响

为研究环保型缓蚀剂壳寡糖对304不锈钢化学机械抛光过程和抛光效果的影响,探讨其在抛光过程中与金属表面的作用方式及吸附机制,采用化学机械抛光试验、接触角测量、扫描电子显微镜(SEM)和X-射线色散能谱仪(EDS)分析等方法,研究壳寡糖有机分子对304不锈钢化学机械抛光的影响,采用量子化学计算研究壳寡糖分子的全局反应参数,分析计算反应活性位点,采用分子动力学模拟有机分子在金属表面的吸附并分析活性原子的径向分布。结果表明：CMP抛光过程中添加壳寡糖能够通过吸附作用在304不锈钢表面形成一层疏水性的薄膜,抑制氧化剂对不锈钢表面的刻蚀,提高抛光后的表面质量;在壳寡糖质量浓度为400 mg/L时得到表面粗糙度为1.65 nm的最佳表面质量。量子化学研究表明,壳寡糖的活性反应位点主要为O原子,能够在金属表面形成多中心吸附。分子动力学模拟表明,壳寡糖有机分子能够平行吸附在金属表面,有机分子中的O原子能够与铁原子形成配位键,在吸附中占据主导地位。     

Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material

Chitin is the most widespread amino renewable carbohydrate polymer in nature and the second most abundant polysaccharide. Therefore, chitin and chitinolytic enzymes are becoming more importance for biotechnological applications in food, health and agricultural fields, the design of effective enzymes being a paramount issue. We report the crystal structure of the plant-type endo-chitinase Chit33 from Trichoderma harzianum and its D165A/E167A-Chit33-(NAG)₄ complex, which showed an extended catalytic cleft with six binding subsites lined with many polar interactions. The major trait of Chit33 is the location of the non-conserved Asp117 and Arg274 acting as a clamp, fixing the distorted conformation of the sugar at subsite –1 and the bent shape of the substrate, which occupies the full catalytic groove. Relevant residues were selected for mutagenesis experiments, the variants being biochemically characterized through their hydrolytic activity against colloidal chitin and other polymeric substrates with different molecular weights and deacetylation percentages. The mutant S118Y stands out, showing a superior performance in all the substrates tested, as well as detectable transglycosylation capacity, with this variant providing a promising platform for generation of novel Chit33 variants with adjusted performance by further design of rational mutants’. The putative role of Tyr in binding was extrapolated from molecular dynamics simulation.