Leonard C G, Ranhand J M, Cole R M
J Bacteriol. 1970 Nov;104(2):674-83. doi: 10.1128/jb.104.2.674-683.1970.
Chemically defined media for competence factor (CF) production by group H Streptococcus strain Challis-6 are described. CF is produced by noncompetent cells in a glutamate-free medium in which the cells cannot attain competence and by cells prior to their competence development in a glutamate-containing medium. Glutamate was required for competence development, but was not necessary for growth or CF production. Exacting cultural conditions required for the consistent production of relatively high amounts of CF in defined medium and for its recovery are detailed. The most important requirements include the selection of isolates (like Challis-6) which grew well in another defined medium, early harvest of CF because of its demonstrated instability on continued incubation in defined medium, incubation at 37 C, and the addition of glucose. The CF production was more rapid with increasing inocula and with reduced aeration. Aspartate, cystine, and NaCl were not required. Under the conditions described, large amounts of CF were consistently obtained in the culture filtrates of Challis-6 as measured by the induction of competence in strain Wicky cells and their subsequent transformation at frequencies of 6% or greater.
本文描述了用于H群链球菌Challis-6菌株产生感受态因子(CF)的化学成分确定的培养基。CF由非感受态细胞在不含谷氨酸的培养基中产生,在这种培养基中细胞无法达到感受态,也由细胞在含谷氨酸的培养基中进入感受态之前产生。谷氨酸是感受态形成所必需的,但对于生长或CF产生并非必需。详细介绍了在限定培养基中持续产生相对大量CF及其回收所需的精确培养条件。最重要的要求包括选择在另一种限定培养基中生长良好的菌株(如Challis-6),由于CF在限定培养基中持续孵育时表现出不稳定性而提前收获,在37℃孵育,以及添加葡萄糖。随着接种量增加和通气减少,CF的产生更快。不需要天冬氨酸、胱氨酸和氯化钠。在所述条件下,通过诱导Wicky菌株细胞的感受态及其随后以6%或更高频率转化来测量,Challis-6的培养滤液中始终能获得大量CF。