Expanding the metabolic engineering toolbox with directed evolution

2013 ◽  
Vol 8 (12) ◽  
pp. 1397-1410 ◽  
Author(s):  
Joseph Abatemarco ◽  
Andrew Hill ◽  
Hal S. Alper
Keyword(s):  
Metabolic Engineering ◽  
Directed Evolution

Metabolic engineering and directed evolution for the production of pharmaceuticals

2000 ◽  
Vol 11 (2) ◽  
pp. 209-214 ◽  
Author(s):  
Michel Chartrain ◽  
Peter M Salmon ◽  
David K Robinson ◽  
Barry C Buckland
Keyword(s):  
Metabolic Engineering ◽  
Directed Evolution

scholarly journals Selection Finder (SelFi): A computational metabolic engineering tool to enable directed evolution of enzymes

2017 ◽  
Vol 4 ◽  
pp. 37-47 ◽  
Author(s):  
Neda Hassanpour ◽  
Ehsan Ullah ◽  
Mona Yousofshahi ◽  
Nikhil U. Nair ◽  
Soha Hassoun
Keyword(s):  
Metabolic Engineering ◽  
Directed Evolution

Metabolic engineering and protein directed evolution increase the yield ofL-phenylalanine synthesized from glucose inEscherichia coli

2004 ◽  
Vol 87 (4) ◽  
pp. 516-524 ◽  
Author(s):  
José Luis Báez-Viveros ◽  
Joel Osuna ◽  
Georgina Hernández-Chávez ◽  
Xavier Soberón ◽  
Francisco Bolívar ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Directed Evolution ◽  
Inescherichia Coli

Directed evolution of adenylosuccinate synthetase from Bacillus subtilis and its application in metabolic engineering

2016 ◽  
Vol 231 ◽  
pp. 115-121 ◽  
Author(s):  
Xiaoyue Wang ◽  
Guanglu Wang ◽  
Xinli Li ◽  
Jing Fu ◽  
Tao Chen ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Engineering ◽  
Directed Evolution ◽  
Adenylosuccinate Synthetase

Metabolic engineering of CHO cells to prepare glycoproteins

2018 ◽  
Vol 2 (3) ◽  
pp. 433-442 ◽  
Author(s):  
Qiong Wang ◽  
Michael J. Betenbaugh
Keyword(s):  
Metabolic Engineering ◽  
Cho Cells ◽  
Genetic Manipulation ◽  
Chinese Hamster ◽  
Post Translational Modification ◽  
Effector Functions ◽  
Recombinant Glycoproteins ◽  
Production Platform ◽  
Chemical Inhibitors ◽  
Amino Acid Mutations

As a complex and common post-translational modification, N-linked glycosylation affects a recombinant glycoprotein's biological activity and efficacy. For example, the α1,6-fucosylation significantly affects antibody-dependent cellular cytotoxicity and α2,6-sialylation is critical for antibody anti-inflammatory activity. Terminal sialylation is important for a glycoprotein's circulatory half-life. Chinese hamster ovary (CHO) cells are currently the predominant recombinant protein production platform, and, in this review, the characteristics of CHO glycosylation are summarized. Moreover, recent and current metabolic engineering strategies for tailoring glycoprotein fucosylation and sialylation in CHO cells, intensely investigated in the past decades, are described. One approach for reducing α1,6-fucosylation is through inhibiting fucosyltransferase (FUT8) expression by knockdown and knockout methods. Another approach to modulate fucosylation is through inhibition of multiple genes in the fucosylation biosynthesis pathway or through chemical inhibitors. To modulate antibody sialylation of the fragment crystallizable region, expressions of sialyltransferase and galactotransferase individually or together with amino acid mutations can affect antibody glycoforms and further influence antibody effector functions. The inhibition of sialidase expression and chemical supplementations are also effective and complementary approaches to improve the sialylation levels on recombinant glycoproteins. The engineering of CHO cells or protein sequence to control glycoforms to produce more homogenous glycans is an emerging topic. For modulating the glycosylation metabolic pathways, the interplay of multiple glyco-gene knockouts and knockins and the combination of multiple approaches, including genetic manipulation, protein engineering and chemical supplementation, are detailed in order to achieve specific glycan profiles on recombinant glycoproteins for superior biological function and effectiveness.

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