An Overview of Heterologous Expression Host Systems for the Production of Recombinant Proteins

Abstract

M atrix metalloproteinases (MMPs) encompasses a family of secreted, calcium dependent, zinc containing endopeptidases. These enzymes are responsible for tissue remodelling and are capable of degrading all kinds of extracellular matrix proteins including collagens, matrix glycoproteins, gelatin, elastins, and proteoglycans. MMPs are involved in regulation of cell behaviours like cellular differentiation, proliferation, angiogenesis, invasion, and apoptosis. Thus, these enzymes have multi-factorial roles during tumor progression: promoting establishment, tumor cell exfoliation, invasion and angiogenesis. The degradation of extracellular matrix components is crucial for tumor cell invasion and MMPs, especially MMP2 (72 kDa gelatinase A) and MMP9 (92 kDa gelatinase B), play critical role in degradation of gelatin and type IV collagen, the two major constituents of extracellular matrix. Short CommunicationAbstract | Matrix Metalloproteinases (MMP) 2 & 9 are overexpressed in wide variety of cancers and play important roles in tumour cell proliferation, invasion and metastasis. Tissue and serum levels of MMP2 and MMP9 correlate with disease prognosis. Real-time PCR is emerging as an alternative or supplementary technique to immunohistochemistry (IHC) for sensitive detection of tumour markers in tissues. Therefore, in this study real-time PCR assays were standardized for detecting over-expression of MMP2 and MMP9 in canine mammary tumour (CMT). Primers designed for real-time PCR analysis of MMP2 & MMP9 were specific as indicated by amplification plots and meltcurve analysis of amplicons. These amplicons showed sharply defined melting curves with single peaks at expected melting points. The over-expression of MMP2 and MMP9 was examined in 12 CMT tissues by real-time PCR analysis. Over-expression of MMP2 & MMP9 was observed in 58.3(7/12) and 41.6 % (5/12) cases of CMT respectively. Expression levels of MMP2 were 2.23±0.10 to 29.80±3.5 fold higher and for MMP2, 1.5±0.56 to 22.67±2.35 fold higher in cases of CMT than normal mammary gland biopsy. Approximately 33.3 % (4/12) of CMT tissues showed co-expression of both MMP2 and MMP9. Further studies are required to correlate MMP2 and MMP9 expression levels with CMT prognosis.

Sustained delivery of the angiogenic QK peptide through the use of polyglutamate domains to control peptide release from bone graft materials

“…Moreover, synthetic peptides such as QK are less expensive to synthesize than recombinant proteins, which require host-cell systems to produce. Host-cell systems can also introduce contaminants such as immunogenic cellular byproducts or pathogens (Gomes, Byregowda, Veeregowda, & Vinayagamurthy, 2016). PGM-QK peptides are synthesized using a commercial peptide synthesizer, and can be stored long-term as a lyophilized product.…”

Structural Properties, Production, and Commercialisation of Invertase

“…As a result, additional purification method is not necessary for subsequent characterisation of the recombinant invertase. Besides, it satisfies the economic efficiency and biosafety regulations for human application (Gomes et al 2016). However, several problems like hyperglycosylation, less secretion of recombinant proteins and a low level of expression reduce the potential of S. cerevisiae in biotechnological industries (Mohandesi et al 2016).…”

“…Recently, several studies have used Pichia pastoris for the production of recombinant invertase from plants, Elsholtzia haichowensis, and baker yeast (S. cerevisiae) (Liu et al 2015;Mohandesi et al 2016). The advantages of using P. pastoris include a high production yield (Yesilirmak & Sayers 2009), good glycosylation capability (Gomes et al 2016), and a superior performance for the expression of secreted proteins (Voegele et al 2006). Comparing the heterologous invertases of E. haichowensis and S. cerevisiae, the latter has a slightly higher optimum pH and temperature (4.8 and 60°C, respectively) compared to the former (pH4.0 and 50°C, respectively).…”

Clonagem, expressão heteróloga e caracterização funcional de peptidase de Fusarium oxysporum em Pichia pastoris

“…Tais sistemas contam com eficientes vetores, que possuem além do gene de interesse, sequências promotoras e/ou regulatórias (FERNANDEZ;HOEFFLER,1999). Além destes, outros sistemas de expressão têm sido desenvolvidos e empregados, como as plantas e animais transgênicos (GOMES et al, 2016).…”

“…A expressão heteróloga possibilita a produção de enzimas em larga escala, bem como permite a inclusão de modificações genéticas. A escolha dos sistemas de expressão por espécies microbianas (a bactéria E. coli e a levedura P. Pastoris) é vantajosa por serem seguros e de fácil cultivo (GOMES et al, 2016).…”