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首页> 外文期刊>Comparative biochemistry and physiology, Part B. Biochemistry & molecular biology >Molecular cloning and characterization of a glycosyl hydrolase family 9 cellulase distributed throughout the digestive tract of the cricket Teleogryllus emma
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Molecular cloning and characterization of a glycosyl hydrolase family 9 cellulase distributed throughout the digestive tract of the cricket Teleogryllus emma

机译：em的整个消化道中分布的糖基水解酶家族9纤维素酶的分子克隆和表征

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摘要

A novel enclogenous beta-1,4-endoglucanase (EG) gene belonging to the glycosyl hydrolase family 9 (GHF 9) that is distributed throughout the digestive tract of the cricket Teleogryllus emma was cloned and characterized. This gene, named TeEG-I, consists of eight exons encoding 453 amino acid residues and exists as a single copy in the T. emma genome. TeEG-I possesses all the features, including signature motifs and catalytic domains, of GHF 9 members, sharing high levels of identity with the termite, Mastotermes darwiniensis (64% protein sequence identity), and the cockroach, Panesthia cribrata (62%), GHF 9 cellulases. Recombinant TeEG-I, which is expressed as a 47-kDa polypeptide in baculovirus-infected insect Sf9 cells, showed an optimal pH and temperature of pH 5.0 and 40 degrees C. The K-m and V-max values for digestion of carboxymethyl cellulose were 5.4 mg/ml and 3118.4 U/mg, respectively. Northern and Western blot analyses revealed that TeEG-I is present throughout the digestive tract, which correlated with the TeEG-I distribution and cellulase activity in the digestive tract as assayed by immunofluorescence staining and enzyme activity assay, respectively. These results indicate that TeEG-I is distributed throughout the entire digestive tract of T. emma, suggesting a functional role of enclogenous TeEG-I in a sequential cellulose digestion process throughout the T. emma digestion tract. (c) 2008 Elsevier Inc. All rights reserved.
机译：克隆并鉴定了一种新的，属于糖基水解酶家族9（GHF 9）的新型β-1,4-内切葡聚糖酶（EG）基因，该基因分布在整个板球Tele的消化道中。该基因名为TeEG-1，由八个编码453个氨基酸残基的外显子组成，并以单拷贝形式存在于艾美螺旋体基因组中。 TeEG-1具有GHF 9成员的所有特征，包括标志性基序和催化结构域，与白蚁，达斯曼白蚁（Mastotermes darwiniensis）（蛋白序列同一性为64％）和蟑螂，番石榴（Panethia cribrata）（62％）具有高度的同一性， GHF 9纤维素酶。在杆状病毒感染的昆虫Sf9细胞中表达为47-kDa多肽的重组TeEG-1的最佳pH和温度为pH 5.0和40摄氏度。羧甲基纤维素消化的Km和V-max值为5.4 mg / ml和3118.4 U / mg。 Northern和Western印迹分析表明，在整个消化道中都存在TeEG-1，这与分别通过免疫荧光染色和酶活性测定法测定的TeEG-1的分布和纤维素酶在消化道中的活性有关。这些结果表明，TeEG-1分布在艾美螺旋体的整个消化道中，暗示了封闭的TeEG-1在整个艾默酸杆菌消化道的顺序纤维素消化过程中的功能性作用。 （c）2008 Elsevier Inc.保留所有权利。

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