共 24 条
Expression of chondroitin-4-O-sulfotransferase in Escherichia coli and Pichia pastoris
被引:30
作者:
He, Wenqin
[1
,2
,3
,4
]
Zhu, Yuanyuan
[5
]
Shirke, Abhijeet
[1
,2
,3
,4
]
Sun, Xiaojun
[6
]
Liu, Jian
[7
]
Gross, Richard A.
[1
,2
,3
,4
]
Koffas, Mattheos A. G.
[1
,2
,3
,4
]
Linhardt, Robert J.
[1
,2
,3
,4
]
Li, Ming
[8
]
机构:
[1] Rensselaer Polytech Inst, Ctr Biotechnol & Interdisciplinary Studies, Dept Chem & Chem Biol, Troy, NY 12180 USA
[2] Rensselaer Polytech Inst, Dept Biol, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA
[3] Rensselaer Polytech Inst, Ctr Biotechnol & Interdisciplinary Studies, Dept Chem & Biol Engn, Troy, NY 12180 USA
[4] Rensselaer Polytech Inst, Dept Biomed Engn, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA
[5] Nanjing Forestry Univ, Dept Chem Proc Engn Forest Prod, Nanjing, Jiangsu, Peoples R China
[6] Shandong Univ, Natl Glycoengn Res Ctr, Shandong Prov Key Lab Carbohydrate Chem & Glycobi, State Key Lab Microbial Technol, Jinan 2500100, Shandong, Peoples R China
[7] Univ N Carolina, Eshelman Sch Pharm, Div Chem Biol & Med Chem, Chapel Hill, NC USA
[8] Tianjin Univ Sci & Technol, Coll Biotechnol, Key Lab Ind Fermentat Microbiol, Tianjin, Peoples R China
基金:
美国国家卫生研究院;
美国国家科学基金会;
关键词:
Chondroitin;
Sulfotransferase;
Bacterial expression;
Yeast expression;
Glycosylation;
Kinetics;
MOLECULAR-CLONING;
CHONDROITIN SULFATE;
HEPARAN-SULFATE;
SULFOTRANSFERASES;
BIOSYNTHESIS;
4-O-SULFOTRANSFERASES;
ENZYMES;
PRODUCE;
FAMILY;
D O I:
10.1007/s00253-017-8411-5
中图分类号:
Q81 [生物工程学(生物技术)];
Q93 [微生物学];
学科分类号:
071005 ;
0836 ;
090102 ;
100705 ;
摘要:
Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase, Homo sapiens C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation. In the current study, a C4ST-1 construct was designed and expressed in both Escherichia coli and Pichia pastoris in its non-glycosylated and glycosylated forms. Both constructs showed similar activity albeit different kinetic parameters when acting on a microbially prepared unsulfated chondroitin substrate. Moreover, the glycosylated form of C4ST-1 showed lower stability than the non-glycosylated form.
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页码:6919 / 6928
页数:10
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