Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose
ABSTRACT: Lacto-N-tetraose (LNT) is one of the most important components of human milk oligosaccharides, which has various beneficial health effects. β-Galactosidase is an important enzyme used in dairy processing. The transglycosylation activity of β-galactosidases offers an attractive approach for...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-10-01
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| Series: | Journal of Dairy Science |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022030223002503 |
| _version_ | 1797676378690158592 |
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| author | Ting Li Jing Li Qiaojuan Yan Shaoqing Yang Zhengqiang Jiang |
| author_facet | Ting Li Jing Li Qiaojuan Yan Shaoqing Yang Zhengqiang Jiang |
| author_sort | Ting Li |
| collection | DOAJ |
| description | ABSTRACT: Lacto-N-tetraose (LNT) is one of the most important components of human milk oligosaccharides, which has various beneficial health effects. β-Galactosidase is an important enzyme used in dairy processing. The transglycosylation activity of β-galactosidases offers an attractive approach for LNT synthesis. In this study, we reported for the first time the biochemical characterization of a novel β-galactosidase (LzBgal35A) from Lacticaseibacillus zeae. LzBgal35A belongs to glycoside hydrolases (GH) family 35 and shared the highest identity of 59.9% with other reported GH 35 members. The enzyme was expressed as soluble protein in Escherichia coli. The purified LzBgal35A displayed optimal activity at pH 4.5 and 55°C. It was stable within the pH range of 3.5 to 7.0 and up to 60°C. Moreover, LzBgal35A could catalyze the synthesis of LNT via transferring the galactose residue from o-nitrophenyl-β-galactopyranoside to lacto-N-triose II. Under optimal conditions, the conversion rate of LNT reached 45.4% (6.4 g/L) within 2 h, which was by far the highest yield of LNT synthesized through a β-galactosidase-mediated transglycosylation reaction. This study demonstrated that LzBgal35A has great potential application in LNT synthesis. |
| first_indexed | 2024-03-11T22:28:13Z |
| format | Article |
| id | doaj.art-c50e5cca77e140d88427332fd6700c89 |
| institution | Directory Open Access Journal |
| issn | 0022-0302 |
| language | English |
| last_indexed | 2024-03-11T22:28:13Z |
| publishDate | 2023-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Dairy Science |
| spelling | doaj.art-c50e5cca77e140d88427332fd6700c892023-09-24T05:13:35ZengElsevierJournal of Dairy Science0022-03022023-10-011061066236634Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraoseTing Li0Jing Li1Qiaojuan Yan2Shaoqing Yang3Zhengqiang Jiang4Department of Nutrition and Health, College of Engineering, China Agricultural University, Haidian District, Beijing 100083, PR ChinaKey Laboratory of China National Light Industry and Food Bioengineering, College of Food Science and Nutritional Engineering, China Agricultural University, Haidian District, Beijing 100083, PR ChinaDepartment of Nutrition and Health, College of Engineering, China Agricultural University, Haidian District, Beijing 100083, PR China; College of Food Science and Engineering, Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing University of Finance and Economics, Nanjing 210023, PR ChinaKey Laboratory of China National Light Industry and Food Bioengineering, College of Food Science and Nutritional Engineering, China Agricultural University, Haidian District, Beijing 100083, PR ChinaKey Laboratory of China National Light Industry and Food Bioengineering, College of Food Science and Nutritional Engineering, China Agricultural University, Haidian District, Beijing 100083, PR China; Corresponding authorABSTRACT: Lacto-N-tetraose (LNT) is one of the most important components of human milk oligosaccharides, which has various beneficial health effects. β-Galactosidase is an important enzyme used in dairy processing. The transglycosylation activity of β-galactosidases offers an attractive approach for LNT synthesis. In this study, we reported for the first time the biochemical characterization of a novel β-galactosidase (LzBgal35A) from Lacticaseibacillus zeae. LzBgal35A belongs to glycoside hydrolases (GH) family 35 and shared the highest identity of 59.9% with other reported GH 35 members. The enzyme was expressed as soluble protein in Escherichia coli. The purified LzBgal35A displayed optimal activity at pH 4.5 and 55°C. It was stable within the pH range of 3.5 to 7.0 and up to 60°C. Moreover, LzBgal35A could catalyze the synthesis of LNT via transferring the galactose residue from o-nitrophenyl-β-galactopyranoside to lacto-N-triose II. Under optimal conditions, the conversion rate of LNT reached 45.4% (6.4 g/L) within 2 h, which was by far the highest yield of LNT synthesized through a β-galactosidase-mediated transglycosylation reaction. This study demonstrated that LzBgal35A has great potential application in LNT synthesis.http://www.sciencedirect.com/science/article/pii/S0022030223002503lacto-N-tetraoseβ-galactosidasetransglycosylationhuman milk oligosaccharides |
| spellingShingle | Ting Li Jing Li Qiaojuan Yan Shaoqing Yang Zhengqiang Jiang Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose Journal of Dairy Science lacto-N-tetraose β-galactosidase transglycosylation human milk oligosaccharides |
| title | Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose |
| title_full | Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose |
| title_fullStr | Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose |
| title_full_unstemmed | Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose |
| title_short | Biochemical characterization of a novel β-galactosidase from Lacticaseibacillus zeae and its application in synthesis of lacto-N-tetraose |
| title_sort | biochemical characterization of a novel β galactosidase from lacticaseibacillus zeae and its application in synthesis of lacto n tetraose |
| topic | lacto-N-tetraose β-galactosidase transglycosylation human milk oligosaccharides |
| url | http://www.sciencedirect.com/science/article/pii/S0022030223002503 |
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