Hyperthermophilic Enzyme Modification of Polysaccharide Biopolymers: Interrelationship Between Biocatalysis and Rheology

这个建议建立在一个正在进行的跨学科的努力，以检查水溶性聚合物的酶修饰和由此产生的流变后果之间的相互关系。本项目扩大的范围有以下目标：(1)有前途的野生型和突变的超嗜热半乳糖甘露聚糖酶和半乳糖糖苷酶的生物化学和生物物理特性，以及多糖修饰；(2)研究酶修饰过程中酶在生物聚合物基质内的迁移和分布，以及伴随而来的生物聚合物-酶体系聚集模式的变化；(3)酶活性与半乳甘露聚糖分子结构和流变学变化的相关性。这样的努力，基于这个研究团队的集体专业知识，即多糖系统，高温下的酶学，凝胶和溶液的流变学，将提供对酶作用导致的聚合物修饰的基本和技术问题的全面理解。虽然这里的重点是半乳甘露聚糖，但这项工作的长期影响将扩展到战略性生物催化方法，以提高其他碳水化合物生物聚合物的功效和使用。

This proposal builds on an ongoing interdisciplinary effort to examine the interrelationship between enzymatic modification of water-soluble polymers and the resulting rheological consequences. The expanded scope of this project has the following objectives: (1) biochemical and biophysical characterization of promising wild-type and mutated hyperthermophilic galactomannanases and galactosidases, with respect to polysaccharide modification; (2) investigation of the mobility and distribution of enzymes within the biopolymer matrix during enzymatic modification along with the concomitant changes in the aggregation pattern of the biopolymer-enzyme system; and (3) correlation of enzyme activity to changes in rheology and galactomannan molecular architecture. Such an effort, based on the collective expertise of this research team, i.e., polysaccharide systems, enzymology at elevated temperatures, and the rheology of gels and solutions, will provide a comprehensive understanding of the fundamental and technological issues governing polymer modification resulting from enzyme action. While the focus here is on galactomannans, the long-term impact of this work will extend to strategic biocatalytic approaches to enhance the efficacy and use of other carbohydrate biopolymers.