Production of recombinant enzymes in the marine alga Dunaliella tertiolecta

Georgianna, D. Ryan; Hannon, Michael J.; Marcuschi, Marina; Wu, Shuiqin; Botsch, Kyle; Lewis, Alex; Hyun, James; Mendez, Michael J.; Mayfield, Stephen P.

doi:10.1016/j.algal.2012.10.004

Cited by 74 publications

(43 citation statements)

References 37 publications

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“…Since that time the tools and techniques for chloroplast genetic engineering of C. reinhardtii have advanced significantly [8,16]. More recently, chloroplast transformation has been achieved for other microalgal species, including the green algae Haematococcus pluvialis and Dunaliella tertiolecta [17,18], the red alga Cyanidioschyzon merolae [19] and the diatom Phaeodactylum tricornutum [20]. However, the progress to date in the development of the algal chloroplast as a platform has almost exclusively focused on C. reinhardtii, with over 100 reports in the literature of the production of recombinant proteins in this species.…”

Section: The Algal Chloroplast As a New Bio-factorymentioning

confidence: 99%

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

2018

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The chloroplast of Chlamydomonas reinhardtii and other microalgae represents an attractive new platform for the synthesis of recombinant therapeutics using synthetic biology (synbio) approaches. Transgenes can be designed in silico, assembled from validated DNA parts and inserted at precise and predetermined locations within the chloroplast genome to give stable synthesis of a desired recombinant protein. Numerous recent examples of different therapeutic proteins produced successfully in the C. reinhardtii chloroplast highlight the potential of this green alga as a simple, low-cost and benign host. Furthermore, some of the features of the alga may offer additional advantages over more-established microbial, mammalian or plant-based systems. These include efficient folding and accumulation of the product in the chloroplast; a lack of contaminating toxins or infectious agents; reduced downstream processing requirements; the possibility to make complex therapeutics such as immunotoxins; and the opportunity to use the whole alga as a low-cost oral vaccine. In this paper we review the current status of algal chloroplast engineering with respect to therapeutic proteins. We also consider future advances in synbio tools, together with improvements to recipient strains, which will allow the design of bespoke strains with high levels of productivity.

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Section: The Algal Chloroplast As a New Bio-factorymentioning

confidence: 99%

The algal chloroplast as a synthetic biology platform for production of therapeutic proteins

2018

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“…1,2) Mannan consists of a backbone of β-1,4-linked mannose residues, whereas konjac glucomannan is a polymer of randomly arranged β-1,4-linked glucose and mannose in the ratio of 1:1.6. The backbone of both mannan and konjac glucomannan can be modified by α-1,6-linked galactosyl residues, forming galactomannan and galactoglucomannan.…”

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confidence: 99%

Purification and characterization of β-Mannanase from Reinekea sp. KIT-YO10 with transglycosylation activity

Hakamada

Ohkubo²,

Ohashi

2014

Bioscience, Biotechnology, and Biochemistry

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Marine bacterium Reinekea sp. KIT-YO10 was isolated from the seashore of Kanazawa Port in Japan as a seaweed-degrading bacterium. Homology between KIT-YO10 16S rDNA and the 16S rDNA of Reinekea blandensis and Reinekea marinisedimentorum was 96.4 and 95.4%, respectively. Endo-1,4-β-D-mannanase (β-mannanase, EC 3.2.1.78) from Reinekea sp. KIT-YO10 was purified 29.4-fold to a 21% yield using anion exchange chromatography. The purified enzyme had a molecular mass of 44.3 kDa, as estimated by SDS-PAGE. Furthermore, the purified enzyme displayed high specificity for konjac glucomannan, with no secondary agarase and arginase activity detected. Hydrolysis of konjac glucomannan and locust bean gum yielded oligosaccharides, compatible with an endo mode of substrate depolymerization. The purified enzyme possessed transglycosylation activity when mannooligosaccharides (mannotriose or mannotetraose) were used as substrates. Optimal pH and temperature were determined to be 8.0 and 70 °C, respectively. It showed thermostability at temperatures from 20 to 50 °C and alkaline stability up to pH 10.0. The current enzyme was thermostable and thermophile compared to the β-mannanase of other marine bacteria.

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“…The gene for α-galactosidase was obtained from the fungus Umbelopsis vinacea and was successfully transformed in the chloroplasts and regulated by endogenous D. tertiolecta promoters and UTR elements. The enzyme α-galactosidase has medicinal applications, such as the treatment of Fabry's disease and prevention of digestive disorders (Georgianna et al, 2013).…”

Section: Genetically Modified (Gm) Microalgaementioning

confidence: 99%