Metabolic engineering of Zymomonas moblis for ethylene production from straw hydrolysate

Abstract Background: Biological ethylene production via the ethylene-forming enzyme (EFE) can offer a promising sustainable alternative approach for fossil-based ethylene production. The high stress tolerance of Z. mobilis make it as promising bio-ethylene producer.Results: In this study, Heterologous expression of the efe gene in Z. mobilis successfully converted the non-ethylene producing strain into an ethylene producer. What’s more, we systematically performed the effect of knocking out the competitive metabolic pathway of pyruvate and the addition of nutrients to the medium to improve the ethylene production in Z. mobilis. These optimization pathways and different substrate supplies resulted in higher ethylene productivity (from 1.36 to 12.83 nmol/OD600/ml), which may guide future engineering work on ethylene production in other organisms to further improve ethylene productivity. Meanwhile, we obtained ethylene production of 5.8 nmol/OD600/ml in strain ZM532-efe by using enzymatic hydrolysate of corn straw as the sole carbon source. This is also the first report on the production of ethylene from cellulosic biomass.Conclusions: These results indicate that the engineered Z. mobilis show great potential for production of ethylene from cellulosic biomass in the future.

Download Full-text

Metabolic engineering of CHO cells to prepare glycoproteins

Emerging Topics in Life Sciences ◽

10.1042/etls20180056 ◽

2018 ◽

Vol 2 (3) ◽

pp. 433-442 ◽

Cited By ~ 1

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.

Download Full-text