scholarly journals Enhanced Levels of the Aroma and Flavor Compound S-Linalool by Metabolic Engineering of the Terpenoid Pathway in Tomato Fruits

2001 ◽  
Vol 127 (3) ◽  
pp. 1256-1265 ◽  
Author(s):  
Efraim Lewinsohn ◽  
Fernond Schalechet ◽  
Jack Wilkinson ◽  
Kenji Matsui ◽  
Yaakov Tadmor ◽  
...  

FEBS Letters ◽  
2002 ◽  
Vol 519 (1-3) ◽  
pp. 30-34 ◽  
Author(s):  
Sridhar Dharmapuri ◽  
Carlo Rosati ◽  
Patrizia Pallara ◽  
Riccardo Aquilani ◽  
Florence Bouvier ◽  
...  


2014 ◽  
Vol 31 (5) ◽  
pp. 567-571 ◽  
Author(s):  
Takashi Kawasaki ◽  
Takao Koeduka ◽  
Akifumi Sugiyama ◽  
Kanako Sasaki ◽  
Philip J. Linley ◽  
...  


2014 ◽  
Vol 23 ◽  
pp. 136-144 ◽  
Author(s):  
Isabel Mendoza-Poudereux ◽  
Jesús Muñoz-Bertomeu ◽  
Alicia Navarro ◽  
Isabel Arrillaga ◽  
Juan Segura


2015 ◽  
Vol 2 (2) ◽  
pp. 75-92 ◽  
Author(s):  
Vered Tzin ◽  
◽  
Ilana Rogachev ◽  
Sagit Meir ◽  
Michal Moyal Ben Zvi ◽  
...  


2014 ◽  
Vol 24 ◽  
pp. 107-116 ◽  
Author(s):  
Michael Gutensohn ◽  
Thuong T.H. Nguyen ◽  
Richard D. McMahon ◽  
Ian Kaplan ◽  
Eran Pichersky ◽  
...  


2021 ◽  
Vol 12 ◽  
Author(s):  
Michael Gutensohn ◽  
Laura K. Henry ◽  
Scott A. Gentry ◽  
Joseph H. Lynch ◽  
Thuong T. H. Nguyen ◽  
...  

Terpenoids are a large and diverse class of plant metabolites that also includes volatile mono- and sesquiterpenes which are involved in biotic interactions of plants. Due to the limited natural availability of these terpenes and the tight regulation of their biosynthesis, there is strong interest to introduce or enhance their production in crop plants by metabolic engineering for agricultural, pharmaceutical and industrial applications. While engineering of monoterpenes has been quite successful, expression of sesquiterpene synthases in engineered plants frequently resulted in production of only minor amounts of sesquiterpenes. To identify bottlenecks for sesquiterpene engineering in plants, we have used two nearly identical terpene synthases, snapdragon (Antirrhinum majus) nerolidol/linalool synthase-1 and -2 (AmNES/LIS-1/-2), that are localized in the cytosol and plastids, respectively. Since these two bifunctional terpene synthases have very similar catalytic properties with geranyl diphosphate (GPP) and farnesyl diphosphate (FPP), their expression in target tissues allows indirect determination of the availability of these substrates in both subcellular compartments. Both terpene synthases were expressed under control of the ripening specific PG promoter in tomato fruits, which are characterized by a highly active terpenoid metabolism providing precursors for carotenoid biosynthesis. As AmNES/LIS-2 fruits produced the monoterpene linalool, AmNES/LIS-1 fruits were found to exclusively produce the sesquiterpene nerolidol. While nerolidol emission in AmNES/LIS-1 fruits was 60- to 584-fold lower compared to linalool emission in AmNES/LIS-2 fruits, accumulation of nerolidol-glucosides in AmNES/LIS-1 fruits was 4- to 14-fold lower than that of linalool-glucosides in AmNES/LIS-2 fruits. These results suggest that only a relatively small pool of FPP is available for sesquiterpene formation in the cytosol. To potentially overcome limitations in sesquiterpene production, we transiently co-expressed the key pathway-enzymes hydroxymethylglutaryl-CoA reductase (HMGR) and 1-deoxy-D-xylulose 5-phosphate synthase (DXS), as well as the regulator isopentenyl phosphate kinase (IPK). While HMGR and IPK expression increased metabolic flux toward nerolidol formation 5.7- and 2.9-fold, respectively, DXS expression only resulted in a 2.5-fold increase.



2013 ◽  
pp. 1581-1597
Author(s):  
Giovanna Giovinazzo ◽  
Ilaria Ingrosso ◽  
Marco Taurino ◽  
Angelo Santino


2018 ◽  
Vol 2 (3) ◽  
pp. 433-442 ◽  
Author(s):  
Qiong Wang ◽  
Michael J. Betenbaugh

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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