一种内切几丁质酶突变体及其编码基因和应用
By performing alanine scanning and PyRosetta design on the endochitinase ThChi8, and introducing N197R and N197RCBM14 mutations, the activity and stability of the enzyme were improved, overcoming the shortcomings of natural enzymes in industrial applications and achieving more efficient chitin degradation and biomass conversion.
CN122146669BActive Publication Date: 2026-07-17NANJING AGRICULTURAL UNIVERSITY
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- NANJING AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-05-08
- Publication Date
- 2026-07-17
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Figure CN122146669B_ABST
Abstract
本发明属于酶学工程和计算机辅助生物技术领域，具体涉及一种内切几丁质酶突变体及其编码基因和应用，所述内切几丁质酶突变体的氨基酸序列如SEQ ID NO.4或SEQ ID NO.6所示。为解决天然内切几丁质酶催化效率低、稳定性差的问题，本发明通过对野生型ThChi8进行计算机辅助理性设计，获得N197R和N197RCBM14融合突变体。实验证明，关键突变体N197RCBM14的比活性提高到野生型的268.25%。所述突变体基因可克隆至pET‑29a(+)载体并在大肠杆菌Rosetta（DE3）中表达，其分泌的酶制剂可高效降解几丁质生产几丁寡糖，在生物质转化、农业及医药领域具有广阔应用前景。
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