Protein Complex Production in Alternative Prokaryotic Hosts

Abstract

Research for multiprotein expression in nonconventional bacterial and archaeal expression systems aims to exploit particular properties of "alternative" prokaryotic hosts that might make them more efficient than E. coli for particular applications, especially in those areas where more conventional bacterial hosts traditionally do not perform well. Currently, a wide range of products with clinical or industrial application have to be isolated from their native source, often microorganisms whose growth present numerous problems owing to very slow growth phenotypes or because they are unculturable under laboratory conditions. In those cases, transfer of the gene pathway responsible for synthesizing the product of interest into a suitable recombinant host becomes an attractive alternative solution. Despite many efforts dedicated to improving E. coli systems due to low cost, ease of use, and its dominant position as a ubiquitous expression host model, many alternative prokaryotic systems have been developed for heterologous protein expression mostly for biotechnological applications. Continuous research has led to improvements in expression yield through these non-conventional models, including Pseudomonas, Streptomyces and Mycobacterium as alternative bacterial expression hosts. Advantageous properties shared by these systems include low costs, high levels of secreted protein products and their safety of use, with non-pathogenic strains been commercialized. In addition, the use of extremophilic and halotolerant archaea as expression hosts has to be considered as a potential tool for the production of mammalian membrane proteins such as GPCRs.

Heterologous and Homologous Expression of Proteins from Haloarchaea: Denitrification as Case of Study

“…The nature of these proteins (significant hydrophobic pattern, high content of amino acidic residues, requirements of high salt concentrations to promote protein folding and stability, requirements related to the activity of chaperons involved in their folding, etc.) makes difficult the production of soluble and active forms when using E. coli as the host for gene expression [10]. Some recent studies have combined different strategies to address this problem.…”

Pseudomonas putida KT2440 is HV1 certified, not GRAS

“…Often P. putida KT2440, and sometimes the P. putida species, is described as Generally Regarded as Safe, or GRAS, indicating it is safe to use as a food additive (Nutrition, ) (e.g. (Aparicio et al ., ; Arnfinnsdottir et al ., ; Belda et al ., ; Bojanovic, ; Calero et al ., ; Cesarini et al ., ; Choi et al ., ; Classen and Pietruszka, ; Cook et al ., ; Cornelissen et al ., ; Cuenca et al ., ; Domröse et al ., ; Dvorak and de Lorenzo, ; Escapa et al ., ; Fröhlich et al ., ; Gemperlein et al ., ; Gómez et al ., ; Gong et al ., ; de las Heras and de Lorenzo, ; Jayakody et al ., ; Kahlon, ; Kim et al ., ; Klein et al ., ; Köppen et al ., ; Kusumawardhani et al ., ; Lee and Wendisch, ; Lieder et al ., ; Loeschcke and Thies, ; Martínez et al ., ; Mindt et al ., ; Nikel and de Lorenzo, ; Pandi et al ., ; Pernicova et al ., ; Poblete‐Castro et al ., ; Puchałka et al ., ; Ravi et al ., ; Rühl et al ., ; Sun et al ., ; Taghavi et al ., ; Troeschel et al ., ; Wang et al ., ; Weyens et al ., ; Wierckx et al ., ; Wu et al ., )).…”