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( Ai Hua Deng ),( Guo Qiang Zhang ),( Na Na Shi ),( Jie Wu ),( Fu Ping Lu ),( Ting Yi Wen ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2
A novel protease gene from Bacillus gibsonii, aprBG, was cloned, expressed in B. subtilis, and characterized. High-level expression of aprBG was achieved in the recombinant strain when a junction was present between the promoter and the target gene. The purified recombinant enzyme exhibited similar N-terminal sequences and catalytic properties to the native enzyme, including high affinity and hydrolytic efficiency toward various substrates and a superior performance when exposed to various metal ions, surfactants, oxidants, and commercial detergents. AprBG was remarkably stable in 50% organic solvents and retained 100% activity and stability in 0-4 M NaCl, which is better than the characteristics of previously reported proteases. AprBG was most closely related to the high-alkaline proteases of the subtilisin family with a 57-68% identity. The secretion and maturation mechanism of AprBG was dependent on the enzyme activity, as analyzed by site-directed mutagenesis. Thus, when taken together, the results revealed that the halo-solvent-tolerant protease AprBG displays significant activity and stability under various extreme conditions, indicating its potential for use in many biotechnology applications.
Jiali Cai,Guochuang Chen,Rongrong Jin,Changhui Deng,Shihui Huang,Xuexia Yuan,Gengjia Chen,Jing Zhao,Zhiyong Wang,Hua Ai 한국공업화학회 2019 Journal of Industrial and Engineering Chemistry Vol.76 No.-
Due to the tumor immune escape mechanism, strategies like the utilization of bispecific antibodies wereused to assist immunotherapy. Herein a visible polymer–inorganic hybrid gene vehicle had beenprepared to deliver anti-EpCAM/anti-CD3 encoding minicircle DNA into normal cells thus thesetransfected cells effectively secreted bispecific antibody with bioactivity. In the experiments, thiscompound system exhibited excellent biocompatibility, high transfection efficiency and ultrasensitiveimaging capacity. Furthermore, the transfected cells could be detected under the MR imaging system andthe gene product showed an outstanding immune effect. In conclusion, this nanocomposite showedsynergistic advantages in gene delivery and non-invasive MR imaging.