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利用关键纤维二糖酶Cba2的重组纤维素酶混合物对滤纸进行协同水解。

Synergistic hydrolysis of filter paper by recombinant cellulase cocktails leveraging a key cellobiase, Cba2, of .

作者信息

Siddique Faiza, Hon Lam Edward Kat, Raymond Wong Wan Keung

机构信息

Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.

Green Faith (International) Environmental Technology Ltd, Unit G, 19/F, King Palace Plaza, Kwun Tong, Kowloon, Hong Kong, China.

出版信息

Front Bioeng Biotechnol. 2022 Sep 28;10:990984. doi: 10.3389/fbioe.2022.990984. eCollection 2022.

Abstract

, a Gram-positive cellulolytic bacterium isolated from soil, is capable of producing a complete cellulase complex exhibiting endoglucanase, exoglucanase, and cellobiase activities. Despite the presence of a full complement of all three types of cellulases, samples prepared from both cell lysates and culture media of showed only weak synergistic activities formed among the cellulase components, as reflected by their inefficient performance in filter paper hydrolysis. However, when the five previously characterized recombinant cellobiases of were mixed individually or in different combinations with recombinant enzyme preparations (CenA/Cex) containing an endoglucanase, CenA, and an exoglucanase, Cex, of another species, , the cellulase cocktails exhibited not only much higher but also synergistic activities in filter paper hydrolysis. Among the 5 cellobiases studied, Cba2 was shown to perform 2.8 to 3.8 times better than other homologous isozymes when acting individually with CenA/Cex. More noteworthy is that when Cba2 and Cba4 were added together to the reaction mixture, an even better synergistic effect was achieved. The filter paper activities resulting from Cba2 and Cba4 interacting with CenA/Cex are comparable to those obtained from some commercial fungal cellulase mixtures. To our knowledge, our results represent the first demonstration of synergistic effects on filter paper hydrolysis achieved using recombinant bacterial cellulases.

摘要

从土壤中分离出的革兰氏阳性纤维素分解菌能够产生具有内切葡聚糖酶、外切葡聚糖酶和纤维二糖酶活性的完整纤维素酶复合物。尽管存在所有三种类型纤维素酶的完整组合,但从该菌的细胞裂解物和培养基中制备的样品在滤纸水解中表现不佳,这反映出纤维素酶组分之间仅表现出微弱的协同活性。然而,当将该菌之前鉴定的五种重组纤维二糖酶分别或与另一种菌([菌名])的含有内切葡聚糖酶CenA和外切葡聚糖酶Cex的重组酶制剂(CenA/Cex)以不同组合混合时,纤维素酶混合物在滤纸水解中不仅表现出更高的活性,而且还具有协同活性。在所研究的5种[菌名]纤维二糖酶中,Cba2与CenA/Cex单独作用时,其表现比其他同源同工酶好2.8至3.8倍。更值得注意的是,当将Cba2和Cba4一起添加到反应混合物中时,实现了更好的协同效应。Cba2和Cba4与CenA/Cex相互作用产生的滤纸活性与一些商业真菌纤维素酶混合物的活性相当。据我们所知,我们的结果首次证明了使用重组细菌纤维素酶对滤纸水解产生的协同效应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/83ff/9554474/ffe29398f064/fbioe-10-990984-g001.jpg

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