草酸青霉菌发酵利用甘草药渣等典型中药固废物的抗逆性机制解析

中药固废物中致密的纤维结构包裹大量药物分子，形成特殊抗性屏障，甘草药渣是一类富含黄酮类、三萜类抑菌成分的典型中药固废物，采用抗逆性菌种将其转化为高附加值产品仍处在发展阶段。草酸青霉菌丰富的代谢系统赋予了它卓越的抗逆性和生物转化性能，是生物质工业化转化的理想菌种。本项目采用“解析-强化-机制解析-验证”的策略，旨在阐明草酸青霉菌利用甘草药渣的抗逆性机制。首先，在解析甘草药渣抗性屏障特征的基础上，利用分子模拟和体外反应考察与甘草药渣特征组分转运和代谢相关的关键酶活性，判定草酸青霉菌的抗逆性因素；随后，采用适应性进化技术，强化草酸青霉菌的抗逆性能；进一步通过基于矩阵分解建模的比较基因组学方法，解析高抗逆表型驯化菌株的突变机制，挖掘抗逆性功能基因；最终，通过基因重构，验证草酸青霉菌利用甘草药渣的抗逆性机制，为解决甘草药渣资源利用问题，也为构建甘草药渣等中药固废物高效生物处置细胞工厂奠定基础。

Herbal extract residues have various small molecules wrapped by dense fiber structure, forming a special resistance barrier. Licorice Resource is a kind of herbal extract residue Rich in flavonoids and triterpenoid antibacterial ingredients. Biomass utilization method based on stress-resistant bacteria has been proven promising but is far from being fully-developed. Penicillium oxalicum's rich metabolic system makes the excellent stress resistance and biotransformation capacity. This species has been recognized as an efficient cell factory for biomass utilization. This project adopts the strategy of "analysis-enhancement-mechanism -verification" to clarify the stress resistance mechanism of Penicillium oxalicum using Licorice Resource and other Herbal extract residues. First of all, on the basis of analyzing the characteristics of resistance barrier of Licorice Resource, the catalytic activities of key enzymes related to the transport and metabolism of solid waste characteristic components of Licorice Resource will be investigated by molecular simulation and in vitro reaction. The stress resistance factors of Penicillium oxalicum will be preliminarily identified through this way, and then the adaptive evolution technique will be used to enhance the stress resistance of Penicillium oxalicum. Furthermore, through the combinatorial analysis based on Matrix factorization modeling, analyze the mutation mechanism of high stress-resistant phenotypic domesticated strains and mine stress-resistant functional genes; finally, through gene knockout technology, verify the stress-resistant mechanism of Penicillium oxalicum in the use of Licorice Resource, which will lays foundation for the utilization of Licorice Resource and the construction of a highly efficient biological disposal cell plant for herbal extract residue.