Hudson Debbie A, Sciascia Quentin L, Sanders Rebecca J, Norris Gillian E, Edwards Pat J B, Sullivan Patrick A, Farley Peter C
Institute of Molecular Biosciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
Institute of Fundamental Sciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand.
Microbiology (Reading). 2004 Sep;150(Pt 9):3041-3049. doi: 10.1099/mic.0.27121-0.
Yeast cells of Candida albicans are induced by serum at 37 degrees C to produce germ tubes, the first step in a transition from yeast to hyphal growth. Previously, it has been shown that the active component is not serum albumin but is present in the dialysable fraction of serum. In this study, serum induction of germ-tube formation is shown to occur even in the presence of added exogenous nitrogen sources and is therefore not signalled by nitrogen derepression. The active component in serum was purified by ion-exchange, reverse-phase and size-exclusion chromatography from the dialysable fraction of serum and was identified by NMR to be d-glucose. Enzymic destruction of glucose, using glucose oxidase, demonstrated that d-glucose was the only active component in these fractions. Induction of germ-tube formation by d-glucose required a temperature of 37 degrees C and the pH optimum was between pH 7.0 and 8.0. d-Glucose induced germ-tube formation in a panel of clinical isolates of C. albicans. Although d-glucose is the major inducer in serum, a second non-dialysable, trichloroacetic acid precipitable inducer is also present. However, whereas either 1.4 % (v/v) serum or an equivalent concentration of d-glucose induced 50 % germ-tube formation, the non-dialysable component required a 10-fold higher concentration to induce 50 % germ-tube formation. Serum is, therefore, the most effective induction medium for germ-tube formation because it is buffered at about pH 8.5 and contains two distinct inducers (glucose and a non-dialysable component), both active at this pH.
白色念珠菌的酵母细胞在37摄氏度下受血清诱导产生芽管,这是从酵母生长向菌丝生长转变的第一步。此前已表明,活性成分不是血清白蛋白,而是存在于血清的可透析部分。在本研究中,即使添加了外源氮源,血清诱导芽管形成仍会发生,因此不是由氮去阻遏发出的信号。血清中的活性成分通过离子交换、反相和尺寸排阻色谱从血清的可透析部分中纯化出来,并通过核磁共振鉴定为d-葡萄糖。使用葡萄糖氧化酶对葡萄糖进行酶促破坏,证明d-葡萄糖是这些部分中唯一的活性成分。d-葡萄糖诱导芽管形成需要37摄氏度的温度,最适pH在7.0至8.0之间。d-葡萄糖在一组白色念珠菌临床分离株中诱导芽管形成。虽然d-葡萄糖是血清中的主要诱导剂,但还存在第二种不可透析的、可被三氯乙酸沉淀的诱导剂。然而,1.4%(v/v)的血清或同等浓度的d-葡萄糖可诱导50%的芽管形成,而不可透析成分需要高10倍的浓度才能诱导50%的芽管形成。因此,血清是芽管形成最有效的诱导培养基,因为它在约pH 8.5下缓冲,并含有两种不同的诱导剂(葡萄糖和一种不可透析成分),两者在此pH下均有活性。