Department of Biomaterials Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
Appl Environ Microbiol. 2010 Sep;76(18):6164-70. doi: 10.1128/AEM.00724-10. Epub 2010 Jul 23.
The wood decay basidiomycete Phanerochaete chrysosporium produces a variety of cellobiohydrolases belonging to glycoside hydrolase (GH) families 6 and 7 in the presence of cellulose. However, no inducer of the production of these enzymes has yet been identified. Here, we quantitatively compared the transcript levels of the genes encoding GH family 6 cellobiohydrolase (cel6A) and GH family 7 cellobiohydrolase isozymes (cel7A to cel7F/G) in cultures containing glucose, cellulose, and cellooligosaccharides by real-time quantitative PCR, in order to evaluate the transcription-inducing effect of soluble sugars. Upregulation of transcript levels in the presence of cellulose compared to glucose was observed for cel7B, cel7C, cel7D, cel7F/G, and cel6A at all time points during cultivation. In particular, the transcription of cel7C and cel7D was strongly induced by cellotriose or cellotetraose. The highest level of cel7C transcripts was observed in the presence of cellotetraose, whereas the highest level of cel7D transcripts was found in the presence of cellotriose, amounting to 2.7 x 10(6) and 1.7 x 10(6) copies per 10(5) actin gene transcripts, respectively. These numbers of cel7C and cel7D transcripts were higher than those in the presence of cellulose. In contrast, cellobiose had a weaker transcription-inducing effect than either cellotriose or cellotetraose for cel7C and had little effect in the case of cel7D. These results indicate that cellotriose and cellotetraose, but not cellobiose, are possible natural cellobiohydrolase gene transcription inducers derived from cellulose.
木质腐朽担子菌嗜热侧耳(Phanerochaete chrysosporium)在存在纤维素的情况下产生各种属于糖苷水解酶(GH)家族 6 和 7 的纤维二糖水解酶。然而,尚未鉴定出这些酶产生的诱导剂。在这里,我们通过实时定量 PCR 定量比较了含有葡萄糖、纤维素和纤维寡糖的培养物中编码 GH 家族 6 纤维二糖水解酶(cel6A)和 GH 家族 7 纤维二糖水解酶同工酶(cel7A 至 cel7F/G)的基因的转录水平,以评估可溶性糖的转录诱导作用。与葡萄糖相比,在纤维素存在的情况下,cel7B、cel7C、cel7D、cel7F/G 和 cel6A 的转录水平均上调。在培养过程中的所有时间点,均观察到 cel7C 和 cel7D 的转录被纤维三糖或纤维四糖强烈诱导。在存在纤维四糖的情况下,cel7C 的转录水平最高,而在存在纤维三糖的情况下,cel7D 的转录水平最高,分别达到 10^5 个肌动蛋白基因转录物的 2.7 x 10^6 和 1.7 x 10^6 个拷贝。这些 cel7C 和 cel7D 转录物的数量高于纤维素存在时的数量。相比之下,纤维二糖对 cel7C 的转录诱导作用比纤维三糖或纤维四糖弱,对 cel7D 的影响很小。这些结果表明,纤维三糖和纤维四糖而不是纤维二糖可能是纤维素衍生的天然纤维二糖水解酶基因转录诱导剂。