Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli

Restrepo-Pineda,; Rosiles-BecerrilVargas-Castillo, Diego; Ávila-Barrientos, Luis P.; Luviano, Axel; Sánchez-Puig, Nuria; García‐Hernández, Enrique; Pérez, Nestor O.; Trujillo‐Roldán, Mauricio A.; Valdez‐Cruz, Norma A.

doi:10.1016/j.ejbt.2022.08.004

Cited by 3 publications

(3 citation statements)

References 63 publications

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“…With respect to the production of IBs, as described by several authors, the concentration and quality of these IBs are affected and can be controlled by various environmental and process factors [ 61 , 62 ], such as temperature, pH, agitation speed, dissolved oxygen concentration, and induction time among others [ 38 , 39 , 40 , 63 ]. This is reflected in the differences observed between our results in shake flasks and bioreactors.…”

Section: Discussionmentioning

confidence: 99%

“…This is reflected in the differences observed between our results in shake flasks and bioreactors. In our case, the temperature condition was not modified throughout the fermentation or between the different scales of the process; thus, contrary to cases in which the induction of recombinant protein production was performed by a change in temperature [ 38 , 64 ], this was not a factor to be considered. Even so, considering that it is desirable to obtain IBs with biological activity, we proceeded to perform the cultures at a temperature of 37 °C, which, according to various authors, is considered a thermal stress condition that favors the production of these IBs [ 65 ].…”

Section: Discussionmentioning

confidence: 99%

“…The production of IBs has been studied in shake flasks and also in bioreactors, although in this last case only with the aim of recovering soluble proteins [ 34 ]. Protein aggregation is improved by stress, such as high temperature, pH shifts, or high feeding rate [ 38 , 39 , 40 ], and, consequently, these parameters tend to result in higher yields of IBs. The unregulated nature of shaken flasks, and lack of controlled pH or oxygen, often results in variation in the expression but not in the quality of the IBs [ 34 ].…”

Section: Introductionmentioning

confidence: 99%

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Immunomodulation Evidence of Nanostructured Recombinant Proteins in Salmonid Cells

Torrealba,

López,

Zelada

et al. 2024

Animals

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Recent studies have demonstrated that immune-related recombinant proteins can enhance immune function, increasing host survival against infectious diseases in salmonids. This research evaluated inclusion bodies (IBs) of antimicrobial peptides (CAMPIB and HAMPIB) and a cytokine (IL1βIB and TNFαIB) as potential immunostimulants in farmed salmonids. For this purpose, we produced five IBs (including iRFPIB as a control), and we evaluated their ability to modulate immune marker gene expression of three IBs in the RTS11 cell line by RT–qPCR. Additionally, we characterized the scale-up of IBs production by comparing two different scale systems. The results showed that CAMPIB can increase the upregulation of tnfα, il1β, il8, and il10, HAMPIB significantly increases the upregulation of tnfα, inos, and il10, and IL1βIB significantly upregulated the expression of tnfα, il1β, and cox2. A comparison of IL1βIB production showed that the yield was greater in shake flasks than in bioreactors (39 ± 1.15 mg/L and 14.5 ± 4.08 mg/L), and larger nanoparticles were produced in shake flasks (540 ± 129 nm and 427 ± 134 nm, p < 0.0001, respectively). However, compared with its shake flask counterpart, the IL1βIB produced in a bioreactor has an increased immunomodulatory ability. Further studies are needed to understand the immune response pathways activated by IBs and the optimal production conditions in bioreactors, such as a defined medium, fed-batch production, and mechanical bacterial lysis, to increase yield.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Immunomodulation Evidence of Nanostructured Recombinant Proteins in Salmonid Cells

Torrealba,

López,

Zelada

et al. 2024

Animals

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Design of Thermoresponsive Genetic Controls with Minimal Heat-Shock Response

Chen,

Jiang,

et al. 2024

ACS Synth. Biol.

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As temperature serves as a versatile input signal, thermoresponsive genetic controls have gained significant interest for recombinant protein production and metabolic engineering applications. The conventional thermoresponsive systems normally require the continuous exposure of heat stimuli to trigger the prolonged expression of targeted genes, and the accompanied heat-shock response is detrimental to the bioproduction process. In this study, we present the design of thermoresponsive quorum-sensing (ThermoQS) circuits to make Escherichia coli record transient heat stimuli. By conversion of the heat input into the accumulation of quorum-sensing molecules such as acyl-homoserine lactone derived from Pseudomonas aeruginosa, sustained gene expressions were achieved by a minimal heat stimulus. Moreover, we also demonstrated that we reprogrammed the E. coli Lac operon to make it respond to heat stimuli with an impressive signal-to-noise ratio (S/ N) of 15.3. Taken together, we envision that the ThermoQS systems reported in this study are expected to remarkably diminish both design and experimental expenditures for future metabolic engineering applications.

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A comprehensive study of glucose and oxygen gradients in a scaled-down model of recombinant HuGM-CSF production in thermoinduced Escherichia coli fed-batch cultures

Reynoso-Cereceda,

Valdez-Cruz,

Pérez

et al. 2024

Preparative Biochemistry & Biotechnology

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Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli

Cited by 3 publications

References 63 publications

Immunomodulation Evidence of Nanostructured Recombinant Proteins in Salmonid Cells

Immunomodulation Evidence of Nanostructured Recombinant Proteins in Salmonid Cells

Design of Thermoresponsive Genetic Controls with Minimal Heat-Shock Response

A comprehensive study of glucose and oxygen gradients in a scaled-down model of recombinant HuGM-CSF production in thermoinduced Escherichia coli fed-batch cultures

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