Ahmadpour Fathollah, Yakhchali Bagher
National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
FEMS Microbiol Lett. 2017 Dec 29;364(24). doi: 10.1093/femsle/fnx216.
Many bacteria, including the genus Bacillus, are able to produce proteases (keratinase). In Bacillus, proteases are produced in the stationary phase and initial stages of sporulation. Protease production is coordinated with sporulation in which expression of various genes by different sigma factors manages the transition from the exponential to the stationary phase. In the present study the sigma-F gene of an indigenous Bacillus cereus strain, which is involved in transcription of genes maintaining sporulation, was deleted. The sigma-H gene, whose product activates the genes that function in the zero phase of sporulation and inhibits suppression of protease production, and spo0B genes were expressed in the exponential phase under the control of a sucrose-inducible promoter from the Bacillus sacPA operon. For the first time, an asporogenic strain of B. cereus was generated that produced higher keratinase (390 U compared with the 198 U of the wild-type strain) and protease (450 U compared with the 290 U of the wild-type strain) activities in the exponential growth phase by induction with sucrose. The new strain is promising for production of keratinase for degradation of feather waste to produce feather meal for poultry feed and decrease environmental pollution from the poultry industry.
许多细菌,包括芽孢杆菌属,都能够产生蛋白酶(角蛋白酶)。在芽孢杆菌中,蛋白酶在稳定期和芽孢形成的初始阶段产生。蛋白酶的产生与芽孢形成相协调,其中不同的西格玛因子对各种基因的表达进行调控,从而控制从指数生长期到稳定期的转变。在本研究中,删除了一株本地蜡样芽孢杆菌菌株中参与维持芽孢形成的基因转录的西格玛-F基因。西格玛-H基因的产物可激活在芽孢形成零期起作用的基因并抑制蛋白酶产生的抑制作用,spo0B基因在来自芽孢杆菌sacPA操纵子的蔗糖诱导型启动子的控制下在指数生长期表达。首次通过蔗糖诱导产生了一株蜡样芽孢杆菌无芽孢菌株,该菌株在指数生长期产生了更高的角蛋白酶(与野生型菌株的198 U相比为390 U)和蛋白酶活性(与野生型菌株的290 U相比为450 U)。该新菌株有望用于生产角蛋白酶,以降解羽毛废弃物来生产家禽饲料用羽毛粉,并减少家禽业对环境的污染。