Messner R, Gruber F, Kubicek C P
Institut für Biochemische Technologie und Mikrobiologie, TU Wien, Austria.
J Bacteriol. 1988 Aug;170(8):3689-93. doi: 10.1128/jb.170.8.3689-3693.1988.
A method consisting of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequent detection of endoglucanases by blotting with a polyclonal antibody against endoglucanase I was used to investigate the effect of induction and carbon catabolite derepression on the synthesis of multiple forms of endoglucanase I by Trichoderma reesei. Five forms appeared upon growth on cellulose, whereas four and only two appeared upon growth on lactose (carbon catabolite derepression) and induction by sophorose in a resting cell system, respectively. All endoglucanases detected resembled endoglucanase I in their specificity, since they exhibited no activity toward xylan or paranitrophenyl-beta-D-lactobioside. A small (25-kilodalton) endoglucanase only appeared during growth on cellulose. None of the multiple forms arose by postsecretional modification. The results indicate that sophorose may not be the only compound mediating cellulose induction of the specific endoglucanases in T. reesei.
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法，随后用抗内切葡聚糖酶I的多克隆抗体进行印迹检测内切葡聚糖酶，以此来研究诱导作用和碳分解代谢物阻遏解除对里氏木霉合成多种形式内切葡聚糖酶I的影响。在以纤维素为碳源生长时出现了5种形式，而在以乳糖为碳源生长（碳分解代谢物阻遏解除）以及在静止细胞体系中用槐糖诱导时，分别出现了4种和2种形式。检测到的所有内切葡聚糖酶在特异性方面都类似于内切葡聚糖酶I，因为它们对木聚糖或对硝基苯基-β-D-乳糖二糖苷没有活性。一种小的（25千道尔顿）内切葡聚糖酶仅在以纤维素为碳源生长时出现。多种形式中没有一种是通过分泌后修饰产生的。结果表明，槐糖可能不是介导里氏木霉中特定内切葡聚糖酶纤维素诱导作用的唯一化合物。
