Yao Xinrun, Lin Min, Yan Yongliang, Jiang Shijie, Zhan Yuhua, Su Bodan, Zhou Zhengfu, Wang Jin
College of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621000, China.
National Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Microorganisms. 2025 Feb 28;13(3):552. doi: 10.3390/microorganisms13030552.
Biotechnological research and application of microbial enzyme production have consistently been focal points for scientific inquiry and industrial advancement. In this study, Y4X3 was isolated from saline-alkaline soil in Xinjiang, China. Extracellular enzyme production analysis revealed that Y4X3 can secrete various enzymes, including cellulase, xylanase, protease, and amylase. Sequencing and assembly of the complete genome of this strain revealed a genome size of 4,215,636 bp with 43.51% C + G content, including 4438 coding genes. Genome annotation was performed with databases to predict gene functions in Y4X3, and a variety of genes related to carbohydrate metabolism were identified. A cellulase-encoding gene was subsequently cloned from the genome and heterologously expressed in . The optimum pH and temperature for the purified cellulase Cel5A were 5.0 and 60 °C, respectively. Stability analysis revealed that Cel5A remained stable at pH 5.0-9.0 and 20-60 °C; after 1 h at pH 9.0, the relative enzyme activity still exceeded 60%. Additionally, Cel5A was positively affected by various metal ions and exhibited good tolerance to multiple chemical reagents. The results indicate that Y4X3 has the potential to produce a variety of industrial enzymes and could serve as a promising candidate for more efficient and cost-effective industrial applications; the characterized thermostable and alkali-resistant cellulase Cel5A also has potential applications in biotechnology and industry.
微生物酶生产的生物技术研究与应用一直是科学探索和工业发展的重点。在本研究中，Y4X3是从中国新疆的盐碱土中分离得到的。细胞外酶生产分析表明，Y4X3能够分泌多种酶，包括纤维素酶、木聚糖酶、蛋白酶和淀粉酶。对该菌株的完整基因组进行测序和组装，结果显示基因组大小为4,215,636 bp，C + G含量为43.51%，包含4438个编码基因。利用数据库进行基因组注释，以预测Y4X3中的基因功能，并鉴定出了多种与碳水化合物代谢相关的基因。随后从基因组中克隆了一个编码纤维素酶的基因，并在[具体宿主]中进行了异源表达。纯化后的纤维素酶Cel5A的最适pH值和温度分别为5.0和60℃。稳定性分析表明，Cel5A在pH 5.0 - 9.0和20 - 60℃范围内保持稳定；在pH 9.0条件下处理1小时后，相对酶活性仍超过60%。此外，Cel5A受到多种金属离子的正向影响，并且对多种化学试剂表现出良好的耐受性。结果表明，Y4X3具有生产多种工业酶的潜力，有望成为更高效、更具成本效益的工业应用的候选菌株；所表征的热稳定且耐碱的纤维素酶Cel5A在生物技术和工业中也具有潜在应用价值。
