Quantitative iTRAQ Secretome Analysis of Cellulolytic Thermobifida fusca
Quantitative iTRAQ Secretome Analysis of Cellulolytic Thermobifida fusca
复制标题
DOI：
10.1021/pr901174z
复制
发表时间：
2010-06-01
影响因子：
4.4
通讯作者：
Sze, Siu Kwan
中科院分区：
文献类型：
--
作者：
Adav, Sunil S.;Ng, Chee Sheng;Sze, Siu Kwan
Thermobifida fusca是放线菌纲的一种嗜热细菌，是植物细胞壁的主要降解菌。对T.采用液相色谱-串联质谱（LC-MS/MS）结合高通量同量异位素相对和绝对定量（iTRAQ）技术的蛋白质组学方法，对生长在纤维素、木质素以及纤维素和木质素混合物培养基中的fusca进行了研究。分泌组的iTRAQ定量揭示了独特的胞外酶系统，包括纤维素酶、半纤维素酶、糖苷水解酶、蛋白酶、过氧化物酶和蛋白质转运转运蛋白的离散多酶复合物。当该菌株生长在这些底物条件下，对应于纤维素酶，半纤维素酶和运输蛋白的蛋白质高度上调，而木质素降解D γ P-型过氧化物酶，新的非血红素过氧化物酶，过氧化氢酶，细胞色素c氧化酶，超氧化物歧化酶也被确定。许多蛋白质假定参与木质纤维素水解的表达在这些不同的培养条件下，其中有几个分泌的假设蛋白质，以前没有观察到。
Thermobifida fusca, a thermophilic bacterium belonging to Actinobacteria, is a major degrader of plant cell walls. The protein profiles of the secretome produced by T. fusca grown in cellulose, lignin, and mixture of cellulose and lignin containing culture media, promoting production of respective substrate hydrolyzing enzymes, was explored using a proteomics approach with high throughput isobaric tag for relative and absolute quantification (iTRAQ) technique using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The iTRAQ quantification of the secretome revealed unique extracellular enzyme system, including discrete multienzyme complexes of cellulases, hemicellulases, glycoside hydrolases, proteases, peroxidases, and protein translocating transporter proteins. When the strain was grown in these substrate conditions, proteins corresponding to cellulases, hemicellulases and transport proteins were highly up-regulated, while lignin degrading D gamma P-type peroxidase, novel nonheme peroxidases, catalase, cytochrome-c oxidase, and superoxide dismutase were also identified. Numerous proteins presumed to be involved in lignocellulose hydrolysis were expressed in response to these different culture conditions, and among these were several secreted hypothetical proteins that were not previously observed.