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在嗜热栖热放线菌中,D-半乳糖诱导细胞外β-半乳糖苷酶(Bga1)形成是由半乳糖醇介导的。

Induction of extracellular beta-galactosidase (Bga1) formation by D-galactose in Hypocrea jecorina is mediated by galactitol.

作者信息

Fekete Erzsébet, Karaffa Levente, Kubicek Christian P, Szentirmai Attila, Seiboth Bernhard

机构信息

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.

出版信息

Microbiology (Reading). 2007 Feb;153(Pt 2):507-512. doi: 10.1099/mic.0.2006/001602-0.

Abstract

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.

摘要

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。

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