N-glycolyl chondroitin synthesis using metabolically engineered E. coli
Abstract N-glycolyl chondroitin (Gc-CN) is a metabolite of N -glycolylneuraminic acid (Neu5Gc), a sialic acid that is commonly found in mammals, but not humans. Humans can incorporate exogenous Neu5Gc into their tissues from eating red meat. Neu5Gc cannot be biosynthesized by humans due to an evolut...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2020-08-01
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| Series: | AMB Express |
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| Online Access: | http://link.springer.com/article/10.1186/s13568-020-01084-6 |
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| author | Adeola E. Awofiranye Sultan N. Baytas Ke Xia Abinaya Badri Wenqin He Ajit Varki Mattheos Koffas Robert J. Linhardt |
| author_facet | Adeola E. Awofiranye Sultan N. Baytas Ke Xia Abinaya Badri Wenqin He Ajit Varki Mattheos Koffas Robert J. Linhardt |
| author_sort | Adeola E. Awofiranye |
| collection | DOAJ |
| description | Abstract N-glycolyl chondroitin (Gc-CN) is a metabolite of N -glycolylneuraminic acid (Neu5Gc), a sialic acid that is commonly found in mammals, but not humans. Humans can incorporate exogenous Neu5Gc into their tissues from eating red meat. Neu5Gc cannot be biosynthesized by humans due to an evolutionary mutation and has been implicated in causing inflammation causing human diseases, such as cancer. The study Neu5Gc is important in evolutionary biology and the development of potential cancer biomarkers. Unfortunately, there are several limitations to detecting Neu5Gc. The elimination of Neu5Gc involves a degradative pathway leading to the incorporation of N-glycolyl groups into glycosaminoglycans (GAGs), such as Gc-CN. Gc-CN has been found in humans and in animals including mice, lamb and chimpanzees. Here, we present the biosynthesis of Gc-CN in bacteria by feeding chemically synthesized N-glycolylglucosamine to Escherichia coli. A metabolically engineered strain of E. coli K4, fed with glucose supplemented with GlcNGc, converted it to N-glycolylgalactosamine (GalNGc) that could then be utilized as a substrate in the chondroitin biosynthetic pathway. The final product, Gc-CN was converted to disaccharides using chondroitin lyase ABC and analyzed by liquid chromatography–tandem mass spectrometry with multiple reaction monitoring detection. This analysis showed the incorporation of GalNGc into the backbone of the chondroitin oligosaccharide. |
| first_indexed | 2024-12-10T22:02:43Z |
| format | Article |
| id | doaj.art-e35747b0d7fc458284e207ed8b6585b4 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-12-10T22:02:43Z |
| publishDate | 2020-08-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-e35747b0d7fc458284e207ed8b6585b42022-12-22T01:31:51ZengSpringerOpenAMB Express2191-08552020-08-011011910.1186/s13568-020-01084-6N-glycolyl chondroitin synthesis using metabolically engineered E. coliAdeola E. Awofiranye0Sultan N. Baytas1Ke Xia2Abinaya Badri3Wenqin He4Ajit Varki5Mattheos Koffas6Robert J. Linhardt7Department of Biological Sciences, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteGlycobiology Research and Training Center, University of CaliforniaDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic InstituteAbstract N-glycolyl chondroitin (Gc-CN) is a metabolite of N -glycolylneuraminic acid (Neu5Gc), a sialic acid that is commonly found in mammals, but not humans. Humans can incorporate exogenous Neu5Gc into their tissues from eating red meat. Neu5Gc cannot be biosynthesized by humans due to an evolutionary mutation and has been implicated in causing inflammation causing human diseases, such as cancer. The study Neu5Gc is important in evolutionary biology and the development of potential cancer biomarkers. Unfortunately, there are several limitations to detecting Neu5Gc. The elimination of Neu5Gc involves a degradative pathway leading to the incorporation of N-glycolyl groups into glycosaminoglycans (GAGs), such as Gc-CN. Gc-CN has been found in humans and in animals including mice, lamb and chimpanzees. Here, we present the biosynthesis of Gc-CN in bacteria by feeding chemically synthesized N-glycolylglucosamine to Escherichia coli. A metabolically engineered strain of E. coli K4, fed with glucose supplemented with GlcNGc, converted it to N-glycolylgalactosamine (GalNGc) that could then be utilized as a substrate in the chondroitin biosynthetic pathway. The final product, Gc-CN was converted to disaccharides using chondroitin lyase ABC and analyzed by liquid chromatography–tandem mass spectrometry with multiple reaction monitoring detection. This analysis showed the incorporation of GalNGc into the backbone of the chondroitin oligosaccharide.http://link.springer.com/article/10.1186/s13568-020-01084-6Sialic acidBiotransformationN-glycolyl chondroitinMetaboliteN-glycolyl glucosamine |
| spellingShingle | Adeola E. Awofiranye Sultan N. Baytas Ke Xia Abinaya Badri Wenqin He Ajit Varki Mattheos Koffas Robert J. Linhardt N-glycolyl chondroitin synthesis using metabolically engineered E. coli AMB Express Sialic acid Biotransformation N-glycolyl chondroitin Metabolite N-glycolyl glucosamine |
| title | N-glycolyl chondroitin synthesis using metabolically engineered E. coli |
| title_full | N-glycolyl chondroitin synthesis using metabolically engineered E. coli |
| title_fullStr | N-glycolyl chondroitin synthesis using metabolically engineered E. coli |
| title_full_unstemmed | N-glycolyl chondroitin synthesis using metabolically engineered E. coli |
| title_short | N-glycolyl chondroitin synthesis using metabolically engineered E. coli |
| title_sort | n glycolyl chondroitin synthesis using metabolically engineered e coli |
| topic | Sialic acid Biotransformation N-glycolyl chondroitin Metabolite N-glycolyl glucosamine |
| url | http://link.springer.com/article/10.1186/s13568-020-01084-6 |
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