Watanabe T, Tanabe T, Sato H, Fuse Y, Ueda K, Nakano K, Saito H, Maehara N
Department of Veterinary Microbiology, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan.
Microbiol Immunol. 1995;39(6):369-77. doi: 10.1111/j.1348-0421.1995.tb02215.x.
Optimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined. High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S. hyicus. The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6. shET activity in the culture filtrate from TY broth inoculated with 10(7) CFU of S. hyicus per ml was higher than that in TY broth inoculated with 10(6) and 10(8) CFU of bacteria per ml. When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions. shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor. shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask. shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose. Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 x 10(9) CFU of S. hyicus strain P-1 into 300 ml of TY broth in a 2,000-ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了猪葡萄球菌产生剥脱毒素(shET)的最佳培养条件。用猪葡萄球菌接种HI和TY肉汤,从其培养滤液中获得了高shET活性。在细菌培养过程中,这两种培养基的pH值在7至8.5之间,而TS和BHI肉汤中的最低pH值小于6。每毫升接种10⁷CFU猪葡萄球菌的TY肉汤培养滤液中的shET活性高于每毫升接种10⁶和10⁸CFU细菌的TY肉汤。当在各种振荡条件下测量培养滤液中的shET活性时,在每分钟75次振荡下振荡的培养滤液在五种振荡条件下具有最高的shET活性。添加了蛋白酶抑制剂的TY肉汤培养滤液的shET活性与未添加抑制剂的TY肉汤相同。在锥形瓶中振荡培养的shET活性也与袋式培养以及使用振荡(坂口)烧瓶振荡培养的活性相同。添加100 mM葡萄糖的TY肉汤中的shET活性明显低于未添加葡萄糖的TY肉汤。基于上述结果,产生shET的最佳培养条件如下:将3×10⁹CFU的猪葡萄球菌菌株P-1接种到2000毫升锥形瓶中的300毫升TY肉汤中,并在37℃下以每分钟75次振荡培养。(摘要截断于250字)