Strategies for humanizing glycosylation pathways and producing recombinant glycoproteins in microbial expression systems

Abstract

Homogeneously glycosylated proteins are essential for analyzing the structure of N‐glycans, studying their consequences inside cells, and developing therapeutic glycoproteins. However, the isolation of glycoproteins with homogeneous glycans from human is difficult since glycoforms slightly differ from each other with respect to molecular weight and charge. Microbial expression systems have numerous benefits in expression technology and have gained great attention, because they are more adaptable to the biotechnology industry. While selecting an expression host, the glycosylation pattern must be taken into account, because glycosylation strongly depends on cellular production system and selected production clone. This review discussed the technological developments in glycoengineering of microbial expression systems for humanizing the glycosylation profile and highlighted the expression potential of Leishmania expression system. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2752, 2019.

An important polysaccharide from fermentum

“…Finally, the Fermentum itself is a high-quality source of nutrition. Vaccines produced with Fermentum cells are called recombinant Fermentum vaccines, and recombinant Fermentum vaccines have been used for a long time to prevent hepatitis B virus (HBV) and human papillomavirus (HPV) ( Khan & Noordin, 2019 ). Liu et al designed an antigen delivery system formed by combining aluminium salts and Fermentum β-polysaccharide particles, and the hepatitis B vaccine prepared from these aluminium salts and Fermentum β-polysaccharide particles was able to promote antigen cross-presentation after subcutaneous injection, with excellent targeting ability.…”

Predicted N‐terminal N‐linked glycosylation sites may underlie membrane protein expression patterns in Saccharomyces cerevisiae

“…We postulate that progression through the secretory pathway and subsequent subcellular and plasma membrane trafficking are strongly influenced by N ‐linked glycosylation at the N‐terminus of membrane proteins in S. cerevisiae . Several reports directly address the link between N ‐linked glycosylation and resulting patterns of expression and trafficking in S. cerevisiae (Khan & Noordin, 2019). This leads us to emphasize the growing importance of understanding this process and its impact in S. cerevisiae , P. pastoris ( K. phaffii ), and other strains of emerging interest, as yeast continues to demonstrate its remarkable use as a model microbe in biotechnology and research applications.…”

“…Another strategy that can be utilized to circumvent aberrant glycosylation patterns is through the use of engineered humanized yeast strains. Efforts have been directed toward humanizing both S. cerevisiae and P. pastoris ( K. phaffii ) whereby glycosylation pathways have been engineered to produce more human‐like patterns (Chiba & Jigami, 2007; Gupta & Shukla, 2018; Jigami, 2008; Khan & Noordin, 2019; Laurent et al, 2016; Wildt & Gerngross, 2005). These efforts have been met with some success; however, optimizing the process still remains somewhat empirical and there are no universally accepted approaches.…”

“…The trade‐off, is lower overall protein production. Regardless, however, this technology may still prove to be more useful compared to other technologies and engineered strains (Gupta & Shukla, 2018; Jacobs et al, 2009; Khan & Noordin, 2019).…”