Extracytoplasmic polysaccharides control cellulosomal and non-cellulosomal systems in Herbivorax saccincola A7

胞质外多糖控制 Herbivorax saccincola A7 中的纤维素体和非纤维素体系统

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作者:Sirilak Baramee, Pakinee Thianheng, Ayaka Uke, Pattsarun Cheawchanlertfa, Chakrit Tachaapaikoon, Rattiya Waeonukul, Patthra Pason, Khanok Ratanakhanokchai, Ya-Jun Liu, Akihiko Kosugi

Abstract

Herbivorax saccincola A7 is an anaerobic alkali-thermophilic lignocellulolytic bacterium that possesses a cellulosome and high xylan degradation ability. To understand the expression profile of extracellular enzymes by carbon sources, quantitative real-time PCR was performed on all cellulosomal and non-cellulosomal enzyme genes of H. saccincola A7 using cellulose and xylan as carbon sources. The results confirmed that the scaffolding proteins of H. saccincola A7 were expressed. In general, the cellulosomal genes belonging to the glycoside hydrolase families 9, 10, 11, and 48 were repressed when xylan was the sole carbon source, but these genes were significantly induced in the presence of cellulose. These results indicate that cellulose, not xylan, is a key inducer of cellulosomal genes in H. saccincola A7. The RsgI-like proteins, which regulate a carbohydrate-sensing mechanism in Clostridium thermocellum, were also found to be encoded in the H. saccincola A7 genome. To confirm the regulation by RsgI-like proteins, the relative expression of σI1-σI4 factors was analyzed on both carbon sources. The expression of alternative σI1 and σI2 factors was enhanced by the presence of cellulose. By contrast, the expression of σI3 and σI4 factors was activated by both cellulose and xylan. Taken together, the results reveal that the cellulosomal and non-cellulosomal genes of H. saccincola A7 are regulated through a carbohydrate-sensing mechanism involving anti-σ regulator RsgI-like proteins. KEY POINTS: • qRT-PCR performed on cellulosomal and non-cellulosomal genes of H. saccincola A7 • Cellulose is a key inducer of the cellulosome of H. saccincola A7 • H. saccincola A7 possesses a similar system of anti-σ regulator RsgI-like proteins.

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