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Abstract：
The thermostable protease TTHA0724 derived from Thermus thermophilus HB8 is an ideal industrial washing enzyme due to its thermophilic characteristics; although it can be expressed in Escherichia coli via pET-22b, high yields are difficult to achieve, leading to frequent autolysis of the host. This paper details the development of a signal peptide library in the expression system of B. subtilis and the optimization of signal peptides for enhanced extracellular expression of TTHA0724. When B. subtilis was used as the host and the optimized signal peptide was used, the expression level of TTHA0724 was 16.7 times higher compared with E. coli. B. subtilis as an expression host does not change the characteristics of TTHA0724. The potential application fields of TTHA0724 are studied. TTHA0724 can be used as a detergent additive at 60 °C, which can sterilize and eliminate mites while thoroughly cleaning protein stains. Soybean meal enzymatic hydrolysis with TTHA0724 at a high temperature produced a higher content of antioxidant peptides. These results indicate that TTHA0724 has great potential for industrial applications.
摘要：
源自嗜热热菌HB8的热稳定蛋白酶TTHA0724由于其嗜热特性，是一种理想的工业洗涤酶；尽管它可以通过pET-22b在大肠杆菌中表达，高产量难以实现，导致宿主频繁的自溶。本文详细介绍了枯草芽孢杆菌表达系统中信号肽库的开发以及用于增强TTHA0724细胞外表达的信号肽的优化。当使用枯草芽孢杆菌作为宿主并使用优化的信号肽时，与大肠杆菌相比，TTHA0724的表达水平高16.7倍。枯草芽孢杆菌作为表达宿主不转变TTHA0724的特征。研究了TTHA0724的潜在应用领域。TTHA0724可在60°C下用作洗涤剂添加剂，它可以消毒和消除螨虫，同时彻底清洁蛋白质污渍。用TTHA0724酶解豆粕在高温下产生了较高含量的抗氧化肽。这些结果表明，TTHA0724具有巨大的工业应用潜力。
