Seiboth B, Hakola S, Mach R L, Suominen P L, Kubicek C P
Abteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien, Vienna, Austria.
J Bacteriol. 1997 Sep;179(17):5318-20. doi: 10.1128/jb.179.17.5318-5320.1997.
The relative contributions of four major cellulases of Trichoderma reesei (1,4-beta-D-glucan cellobiohydrolase I [CBH I], CBH II, endo-1,4-beta-D-glucanase I [EG I], and EG II) to the generation of the cellulase inducer from cellulose were studied with isogenic strains in which the corresponding genes (cbh1, cbh2, egl1, and egl2) had been deleted by insertion of the Aspergillus nidulans amdS marker gene. During growth on lactose (a soluble carbon source provoking cellulase gene expression), these strains showed no significant alterations in their ability to express the respective other cellulase genes, with the exception of the strain containing delta cbh1, which exhibited an increased steady-state level of cbh2 mRNA. On crystalline cellulose as the only carbon source, however, significant differences were apparent: strains in which cbh2 and egl2, respectively, had been deleted showed no expression of the other cellulase genes, whereas strains carrying the cbh1 or egl1 deletion showed these transcripts. The delta cbh1-containing strain also showed enhanced cbh2 mRNA levels under these conditions. A strain in which both cbh1 and cbh2 had been deleted, however, was unable to initiate growth on cellulose. Addition of 2 mM sophorose, a putative inducer of cellulase gene expression, to such cultures induced the transcription of egl1 and egl2 and restored the ability to grow on cellulose. We conclude that CBH II and EG II are of major importance for the efficient formation of the inducer from cellulose in T. reesei and that removal of both cellobiohydrolases renders T. reesei unable to attack crystalline cellulose.
利用同源菌株研究了里氏木霉的四种主要纤维素酶(1,4-β-D-葡聚糖纤维二糖水解酶I [CBH I]、CBH II、内切-1,4-β-D-葡聚糖酶I [EG I]和EG II)对从纤维素生成纤维素酶诱导物的相对贡献,在这些同源菌株中,相应基因(cbh1、cbh2、egl1和egl2)已通过插入构巢曲霉amdS标记基因而缺失。在乳糖(一种可引发纤维素酶基因表达的可溶性碳源)上生长期间,这些菌株在表达各自其他纤维素酶基因的能力方面没有显著变化,但含有δcbh1的菌株除外,该菌株显示cbh2 mRNA的稳态水平有所增加。然而,以结晶纤维素作为唯一碳源时,明显存在显著差异:分别缺失cbh2和egl2的菌株没有其他纤维素酶基因的表达,而携带cbh1或egl1缺失的菌株则有这些转录本。在这些条件下,含有δcbh1的菌株也显示出cbh2 mRNA水平升高。然而,一种同时缺失cbh1和cbh2的菌株无法在纤维素上起始生长。向此类培养物中添加2 mM槐糖(一种假定的纤维素酶基因表达诱导物)可诱导egl1和egl2的转录,并恢复在纤维素上生长的能力。我们得出结论,CBH II和EG II对于里氏木霉从纤维素高效形成诱导物至关重要,并且去除这两种纤维二糖水解酶会使里氏木霉无法分解结晶纤维素。