Directed evolution of a β-N-acetylhexosaminidase from Haloferula sp. for lacto-N-triose II and lacto-N-neotetraose synthesis from chitin.
Autor: | Liu Y; Key Laboratory of Food Bioengineering (China National Light Industry), College of Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China., Yan Q; Key Laboratory of Food Bioengineering (China National Light Industry), College of Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China., Ma J; Key Laboratory of Food Bioengineering (China National Light Industry), College of Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China., Wang J; Key Laboratory of Food Bioengineering (China National Light Industry), College of Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China., Jiang Z; Department of Nutrition and Health, College of Food Science and Nutritional Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China. Electronic address: zhqjiang@cau.edu.cn., Yang S; Department of Nutrition and Health, College of Food Science and Nutritional Engineering, China Agricultural University, No.17 Qinghua East Road, Haidian, Beijing 100083, China. Electronic address: ysq@cau.edu.cn. |
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Jazyk: | angličtina |
Zdroj: | Enzyme and microbial technology [Enzyme Microb Technol] 2023 Mar; Vol. 164, pp. 110177. Date of Electronic Publication: 2022 Dec 15. |
DOI: | 10.1016/j.enzmictec.2022.110177 |
Abstrakt: | In our previous study, a β-N-acetylhexosaminidase (HaHex74) from Haloferula sp. showing high human milk oligosaccharides (HMOs) synthesis ability was identified and characterized. In this study, HaHex74 was further engineered by directed evolution and site-saturation mutagenesis to improve its transglycosylation activity for HMOs synthesis. A mutant (mHaHex74) with improved transglycosylation activity (HaHex74-Asn401Ile/His394Leu) was obtained and characterized. mHaHex74 exhibited maximal activity at pH 5.5 and 35 °C, respectively, which were distinct from that of HaHex74 (pH 6.5 and 45 °C). Moreover, mHaHex74 showed the highest LNT2 conversion ratio of 28.2% from N,N'-diacetyl chitobiose (GlcNAc Competing Interests: Declaration of Competing Interest The authors declare that there are no conflict of interest in the manuscript. (Copyright © 2022 Elsevier Inc. All rights reserved.) |
Databáze: | MEDLINE |
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