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在不同底物上生长的琥珀酸纤维杆菌S85的内切葡聚糖酶活性及糖苷水解酶基因的相对表达。

Endoglucanase activity and relative expression of glycoside hydrolase genes of Fibrobacter succinogenes S85 grown on different substrates.

作者信息

Béra-Maillet C, Gaudet G, Forano E

机构信息

Laboratoire de Microbiologie, INRA, Centre de Clermont-Ferrand-Theix, 63122, Saint-Genès-Champanelle, France.

出版信息

Biochim Biophys Acta. 2000 Nov 30;1543(1):77-85. doi: 10.1016/s0167-4838(00)00194-1.

Abstract

The endoglucanase activity of cells and extracellular culture fluid of Fibrobacter succinogenes S85 grown on glucose, cellobiose, soluble polysaccharides (beta-glucan, lichenan) and intact plant polysaccharides, was compared. The specific activity of cells grown on cellulose or forages was 6- to 20-fold higher than that of cells grown on soluble substrates, suggesting an induction of endoglucanases by the insoluble substrates. The ratios of cells to extracellular culture fluid endoglucanase activities measured in cultures grown on sugars or insoluble polysaccharides suggested that the endoglucanases induced by the insoluble polysaccharides remained attached to the cells. The mRNA of all the F. succinogenes glycoside hydrolase genes sequenced so far were then quantified in cells grown on glucose, cellobiose or cellulose. The results show that all these genes were transcribed in growing cells, and that they are all overexpressed in cultures grown on cellulose. Endoglucanase-encoding endB and endA(FS) genes, and xylanase-encoding xynC gene appeared the most expressed genes in growing cells. EGB and ENDA are thus likely to play a major role in cellulose degradation in F. succinogenes.

摘要

比较了生长在葡萄糖、纤维二糖、可溶性多糖(β-葡聚糖、地衣多糖)和完整植物多糖上的琥珀酸纤维杆菌S85细胞及细胞外培养液的内切葡聚糖酶活性。在纤维素或草料上生长的细胞的比活性比在可溶性底物上生长的细胞高6至20倍,这表明不溶性底物可诱导内切葡聚糖酶的产生。在以糖类或不溶性多糖为培养基生长的培养物中测得的细胞与细胞外培养液内切葡聚糖酶活性的比率表明,由不溶性多糖诱导产生的内切葡聚糖酶仍附着在细胞上。然后对迄今已测序的所有琥珀酸纤维杆菌糖苷水解酶基因的mRNA在以葡萄糖、纤维二糖或纤维素为培养基生长的细胞中进行定量分析。结果表明,所有这些基因在生长的细胞中均有转录,并且在以纤维素为培养基生长的培养物中均过度表达。编码内切葡聚糖酶的endB和endA(FS)基因以及编码木聚糖酶的xynC基因似乎是生长细胞中表达最多的基因。因此,EGB和ENDA可能在琥珀酸纤维杆菌降解纤维素的过程中起主要作用。

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