Glycoprotein In Vitro N-Glycan Processing Using Enzymes Expressed in E. coli
Abstract
Protein N-glycosylation is a common post-translational modification that plays significant roles on the structure, property, and function of glycoproteins. Due to N-glycan heterogeneity of naturally occurring glycoproteins, the functions of specific N-glycans on a particular glycoprotein are not always clear. Glycoprotein in vitro N-glycan engineering using purified recombinant enzymes is an attractive strategy to produce glycoproteins with homogeneous N-glycoforms to elucidate the specific functions of N-glycans and develop better glycoprotein therapeutics. Toward this goal, we have successfully expressed in E. coli glycoside hydrolases and glycosyltransferases from bacterial and human origins and developed a robust enzymatic platform for in vitro processing glycoprotein N-glycans from high-mannose-type to α2–6- or α2–3-disialylated biantennary complex type. The recombinant enzymes are highly efficient in step-wise or one-pot reactions. The platform can find broad applications in N-glycan engineering of therapeutic glycoproteins.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 18, 2023
- Source ID
- 10.3390/molecules28062753
Entities
People
- Hai Yu
- Jingxin Fu
- Libo Zhang
- Riyao Li
- Xi Chen
- Xiaohong Yang
- Yanhong Li
- Zimin Zheng
Organizations
- Defense Threat Reduction Agency
- National Institutes of Health
- United States Department of Commerce
- University of California