Fekete Erzsébet, Karaffa Levente, Kubicek Christian P, Szentirmai Attila, Seiboth Bernhard
Department of Genetics and Applied Microbiology, Faculty of Science, University of Debrecen, H-4010, PO Box 56, Debrecen, Hungary.
Research Area Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, TU Wien, Getreidemarkt 9/166-5, A-1060 Wien, Austria.
Microbiology (Reading). 2007 Feb;153(Pt 2):507-512. doi: 10.1099/mic.0.2006/001602-0.
The ability of Hypocrea jecorina (Trichoderma reesei) to grow on lactose strongly depends on the formation of an extracellular glycoside hydrolase (GH) family 35 beta-galactosidase, encoded by the bga1 gene. Previous studies, using batch or transfer cultures of pregrown cells, had shown that bga1 is induced by lactose and d-galactose, but to a lesser extent by galactitol. To test whether the induction level is influenced by the different growth rates attainable on these carbon sources, bga1 expression was compared in carbon-limited chemostat cultivations at defined dilution (=specific growth) rates. The data showed that bga1 expression by lactose, d-galactose and galactitol positively correlated with the dilution rate, and that galactitol and d-galactose induced the highest activities of beta-galactosidase at comparable growth rates. To know more about the actual inducer for beta-galactosidase formation, its expression in H. jecorina strains impaired in the first steps of the two d-galactose-degrading pathways was compared. Induction by d-galactose and galactitol was still found in strains deleted in the galactokinase-encoding gene gal1, which is responsible for the first step of the Leloir pathway of d-galactose catabolism. However, in a strain deleted in the aldose/d-xylose reductase gene xyl1, which performs the reduction of d-galactose to galactitol in a recently identified second pathway, induction by d-galactose, but not by galactitol, was impaired. On the other hand, induction by d-galactose and galactitol was not affected in an l-arabinitol 4-dehydrogenase (lad1)-deleted strain which is impaired in the subsequent step of galactitol degradation. These results indicate that galactitol is the actual inducer of Bga1 formation during growth on d-galactose in H. jecorina.
嗜热栖热菌(里氏木霉)在乳糖上生长的能力很大程度上取决于由bga1基因编码的胞外糖苷水解酶(GH)家族35β-半乳糖苷酶的形成。以往使用预培养细胞的分批培养或转接培养的研究表明,bga1由乳糖和D-半乳糖诱导,但由半乳糖醇诱导的程度较小。为了测试诱导水平是否受这些碳源上可达到的不同生长速率的影响,在定义的稀释(=比生长)速率下,在碳限制恒化器培养中比较了bga1的表达。数据表明,乳糖、D-半乳糖和半乳糖醇对bga1的表达与稀释速率呈正相关,并且在可比生长速率下,半乳糖醇和D-半乳糖诱导的β-半乳糖苷酶活性最高。为了更多地了解β-半乳糖苷酶形成的实际诱导物,比较了其在两条D-半乳糖降解途径第一步受损的嗜热栖热菌菌株中的表达。在缺失编码半乳糖激酶的基因gal1的菌株中,仍发现由D-半乳糖和半乳糖醇诱导,该基因负责D-半乳糖分解代谢的Leloir途径的第一步。然而,在缺失醛糖/D-木糖还原酶基因xyl1的菌株中,该基因在最近确定的第二条途径中负责将D-半乳糖还原为半乳糖醇,由D-半乳糖而非半乳糖醇诱导受损。另一方面,在缺失L-阿拉伯糖醇4-脱氢酶(lad1)的菌株中,半乳糖醇降解的后续步骤受损,由D-半乳糖和半乳糖醇诱导不受影响。这些结果表明,半乳糖醇是嗜热栖热菌在D-半乳糖上生长期间Bga1形成的实际诱导物。