Xie Mengqi, Li Yilin, Xu Luning, Zhang Shusheng, Ye Hongyu, Sun Faqian, Mei Rongwu, Su Xiaomei
College of Geography and Environmental Science, Zhejiang Normal University Yingbin Road 688# Jinhua 321004 China
The Management Center of Wuyanling National Natural Reserve in Zhejiang Wenzhou 325500 China.
RSC Adv. 2021 Nov 9;11(57):36105-36115. doi: 10.1039/d1ra03565g. eCollection 2021 Nov 4.
In natural and engineered systems, most microorganisms would enter a state of dormancy termed as "viable but non-culturable" (VBNC) state when they are exposed to unpredictable environmental stress. One of the major advances in resuscitating from such a state is the discovery of a kind of bacterial cytokine protein called resuscitation-promoting factor (Rpf), which is secreted from . In this study, the optimization of Rpf production was investigated by the response surface methodology (RSM). Results showed that an empirical quadratic model well predicted the Rpf yield, and the highest Rpf protein yield could be obtained at the optimal conditions of 59.56 mg L IPTG, cell density 0.69, induction temperature 20.82 °C and culture time 7.72 h. Importantly, Phyre2 web portal characterized the structure of the Rpf domain to have a shared homology with lysozymes, and the highest lysozyme activity was at pH 5 and 50 °C. This study broadens the knowledge of Rpf production and provided potential strategies to apply Rpf as a bioactivator for environmental bioremediation.
在自然和工程系统中,大多数微生物在受到不可预测的环境压力时会进入一种被称为“活的非可培养”(VBNC)状态的休眠状态。从这种状态复苏的一项重大进展是发现了一种名为复苏促进因子(Rpf)的细菌细胞因子蛋白,它是从……分泌的。在本研究中,采用响应面法(RSM)研究了Rpf产量的优化。结果表明,经验二次模型能很好地预测Rpf产量,在IPTG 59.56 mg/L、细胞密度0.69、诱导温度20.82℃和培养时间7.72 h的最佳条件下可获得最高的Rpf蛋白产量。重要的是,Phyre2门户网站对Rpf结构域的结构进行了表征,发现其与溶菌酶具有共同的同源性,且最高溶菌酶活性在pH 5和50℃时出现。本研究拓宽了对Rpf生产的认识,并提供了将Rpf作为环境生物修复生物激活剂应用的潜在策略。