Production and characterization of pharmacologically active recombinant human phosphodiesterase 4B in <i>Dictyostelium discoideum</i>

Arya, Ranjana; Aslam, Saima; Gupta, Shivani; Bora, Roop Singh; Vijayakrishnan, Lalitha; Gulati, Pankaj; Naithani, Sudha; Mukherjee, Shohini; Dastidar, Sunanda G.; Bhattacharya, Alok; Saini, Kulvinder Singh

doi:10.1002/biot.200700256

Cited by 7 publications

(3 citation statements)

References 52 publications

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“…Furthermore, the same group demonstrated an enhancement in the transfection efficiencies by inhibiting endocytosis after DNA uptake (Vats and Padh, 2009). Arya et al, (2008a; successfully purified human recombinant proteins, PDE4 and PDE7 (both phosphodiestreases), in D. discoideum and similarly human spleen tyrosine kinase and GNE proteins were also purified (Singh et al, 2010;Grover et al, 2014). Future studies should be focused on optimizing heterologous protein production and studying the bioassay of recombinants proteins.…”

Section: Production Of Heterologous Proteinsmentioning

confidence: 97%

Glimpses of Dictyostelid research in India

Begum¹,

Saran²

2020

Int. J. Dev. Biol.

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Simple organisms are preferred for understanding the molecular and cellular function(s) of complex processes. Dictyostelium discoideum is a lower eukaryote, a protist and a cellular slime mould, which has been in recent times used for various studies such as cell differentiation, development, cell death, stress responses etc. It is a soil amoeba (unicellular) that undertakes a remarkable, facultative shift to multicellularity when exposed to starvation and requires signal pathways that result in alteration of gene expression and finally show cell differentiation. The amoebae aggregate, differentiate and form fruiting bodies with two terminally differentiated cells: the dead stalk (non-viable) and dormant spores (viable). In India, starting from the isolation of Dictyostelium species to morphogenesis, cell signalling and social evolution has been studied with many more new research additions. Advances in molecular genetics make Dictyostelium an attractive model system to study cell biology, biochemistry, signal transduction and many more.

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Section: Production Of Heterologous Proteinsmentioning

confidence: 97%

Glimpses of Dictyostelid research in India

Begum¹,

Saran²

2020

Int. J. Dev. Biol.

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“…human antithrombin III; Dingermann et al 1991, Tiltscher and Storr 1993, Dittrich et al 1994) and enzymes (e.g. phosphodiesterase; Arya et al 2008a). Its advantage over other expression systems lies in its extensive post-translational modification system (glycosylation, phosphorylation, acylation), which resembles that of higher eukaryotes (Jung and Williams 1997, Slade et al 1997.…”

Section: Dictyostelium Discoideummentioning

confidence: 99%

Biocatalysis, Enzyme Engineering and Biotechnology

Kotzia

Platis

Axarli

et al. 2012

Food Biochemistry and Food Processing

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Enzymes are biocatalysts evolved in nature to achieve the speed and coordination of nearly all the chemical reactions that define cellular metabolism necessary to develop and maintain life. The application of biocatalysis is growing rapidly, since enzymes offer potential for many exciting applications in industry. The advent of whole genome sequencing projects enabled new approaches for biocatalyst development, based on specialised methods for enzyme heterologous expression and engineering. The engineering of enzymes with altered activity, specificity and stability, using sitedirected mutagenesis and directed evolution techniques are now well established. Over the last decade, enzyme immobilisation has become important in industry. New methods and techniques for enzyme immobilisation allow for the reuse of the catalysts and the development of efficient biotechnological processes. This chapter reviews advances in enzyme technology as well as in the techniques and strategies used for enzyme production, engineering and immobilisation and discuss their advantages and disadvantages. HO CH 2 COOH CH NH 2 Negatively charged amino acids Glutamate Glu (E) CH 2 CH 2 HOOC COOH CH NH 2 Aspartate Asn (N) CH 2 COOH HOOC CH NH 2 Positively charged amino acids Arganine Arg (R) CH 2 CH 2 CH 2 HN COOH CH NH 2 C NH NH 2 Lysine Lys (K) H 2 N COOH (CH 2) 4 CH NH 2 Histidine His (H)

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“…Expression of full‐length PDE isoforms is essential for HTS assays for the identification of PDE subtype selective inhibitors that would offer better efficacy profile over existing molecules. We have successfully expressed and purified active recombinant human PDE4B2 in Dd that showed similar kinetic profile, as compared to PDE4B2 expressed in mammalian cells (97). Recombinant human PDE7A1 was also expressed in a functionally active form in Dd , and up to 2 mg/l of full‐length protein was purified from cell lysates (98).…”

Section: Therapeutically Important Proteins Produced In Dd: Key Lessomentioning

confidence: 99%

Dictyostelium discoideum—a promising expression system for the production of eukaryotic proteins

Arya¹,

Bhattacharya

Saini³

2008

FASEB j.

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In general, four different expression systems, namely, bacterial, yeast, baculovirus, and mammalian, are widely used for the overproduction of biochemical enzymes and therapeutic proteins. Clearly, bacterial expression systems offer ease of maneuverability with respect to large-scale production of recombinant proteins, while, a baculovirus expression system ensures proper protein modifications, processing, and refolding of complex proteins. Despite these advantages, mammalian cells remain the preferred host for many eukaryotic proteins of pharmaceutical importance, particularly, those requiring post-translational modifications. Recently, the single-celled slime mold, Dictyostelium discoideum (Dd), has emerged as a promising eukaryotic host for the expression of a variety of heterologous recombinant eukaryotic proteins. This organism possesses the complex cellular machinery required for orchestrating post-translational modifications similar to the one observed in higher eukaryotes. This review summarizes the advantages and disadvantages of Dictyostelium as an alternate system compared to other well-established expression systems. The key lessons learned from the expression of human recombinant proteins in this system are reviewed. Also, the strengths, weaknesses, and challenges associated with industrial-scale production of proteins in Dd expression system are discussed.

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Production and characterization of pharmacologically active recombinant human phosphodiesterase 4B in Dictyostelium discoideum

Cited by 7 publications

References 52 publications

Glimpses of Dictyostelid research in India

Glimpses of Dictyostelid research in India

Biocatalysis, Enzyme Engineering and Biotechnology

Dictyostelium discoideum—a promising expression system for the production of eukaryotic proteins

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