Evaluation of induction conditions for plasmid pQE-30 stability and 503 antigen of Leishmania i. chagasi expression in E. coli M15

Ribeiro, Vitor Troccoli; Asevedo, Estéfani Alves; Vasconcelos, Luan Tales Costa de Paiva; Filho, Marcos Antônio Oliveira; Araújo, Jaciara Silva de; Macêdo, Gorete Ribeiro de; Júnior, Francisco Canindé de Sousa; Santos, Everaldo Silvino dos

doi:10.1007/s00253-019-09948-z

Cited by 11 publications

(7 citation statements)

References 38 publications

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“…24 More recently, the native Shewanella trimethylamine N-oxide (TMAO)-dependent promoters were also successfully used to regulate gene expression in S. oneidensis MR-1. 25 However, these inducible promoters have several disadvantages that limit their wide applications in Shewanella species, including leaky expression (IPTG-or arabinose-inducible), 26,27 nonlinearity with respect to inducer concentration (IPTG-or arabinoseinducible), toxicity to bacterial cells (IPTG-or arsenicinducible), 28 and low induction efficiency (TMAO-inducible). 29 Therefore, more effective and compatible inducible expression systems for Shewanella species are highly desired.…”

Section: ■ Introductionmentioning

confidence: 99%

Engineering a Rhamnose-Inducible System to Enhance the Extracellular Electron Transfer Ability of Shewanella Genus for Improved Cr(VI) Reduction

Cheng

Min

et al. 2021

ACS EST Eng.

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Members of the Shewanella genus play important roles in bioenergy generation and environmental bioremediation, but their practical applications in engineered systems are still limited due to the lack of effective synthetic biology tools and parts such as promoters and expression systems. In this work, the rhaBAD promoter of Escherichia coli was introduced into various Shewanella species including S. oneidensis MR-1, S. decolorationis S12, and S. aquimarina SW-120 to enhance their extracellular pollutant transformation ability with Cr(VI) reduction as an example. The rhaBAD promoter had low leaky expression without L-rhamnose induction and exhibited a linear response to a wide concentration range. The inducer was nontoxic and could not be metabolizable by the Shewanella species. In addition, a unimodal induction of the rhaBAD promoter was found at the single-cell level. Applying this rhamnose-inducible system to the Shewanella species allowed for precise control on their extracellular electron transfer abilities to promote Cr(VI) transformation. The engineered strain induced by L-rhamnose reduced 98% Cr(VI) within 9 h, while only 59.1% Cr(VI) reduction was achieved by the control strain. Moreover, a 3-fold higher Cr(VI) reduction rate for the engineered strain induced by L-rhamnose confirms the improvement of its biotransformation ability. Our work provides an efficient regulation tool for engineering Shewanella species and may hopefully bring them a step further toward practical applications.

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Section: ■ Introductionmentioning

confidence: 99%

Engineering a Rhamnose-Inducible System to Enhance the Extracellular Electron Transfer Ability of Shewanella Genus for Improved Cr(VI) Reduction

Cheng

Min

et al. 2021

ACS EST Eng.

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“…Regulation of inducer concentration is critical for recombinant protein expression [ 17 , 18 ]. High concentration of inducer can have toxic effects on host cells, thus preventing protein expression.…”

Section: Resultsmentioning

confidence: 99%

Expression and purification of soluble recombinant β-lactamases using Escherichia coli as expression host and pET-28a as cloning vector

Tian

et al. 2022

Microb Cell Fact

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Background Due to its high expression capability, recombination of Escherichia coli and pET vector has become the bioengineering preferred expression system. Because β-lactamases mediate bacterial antimicrobial resistance, these enzymes have a substantial clinical impact. Using the E. coli expression system, several kinds of β-lactamases have been produced. However, previous studies have been focused on characterizing target β-lactamases, and the effects of cultivation and induction conditions on the expression efficiency of target enzymes were not addressed. Results Using pET-28a as the cloning vector and E. coli BL21(DE3) as the expression host, this study originally elucidated the effects of IPTG concentration, culture temperature, induction time, and restriction sites on recombinant β-lactamase expression. Moreover, the effects of the target protein length and the 6 × His-tag fusion position on enzyme purification were also explored, and consequently, this study yielded several important findings. (i) Only the signal peptide–detached recombinant β-lactamase could exist in a soluble form. (ii) Low-temperature induction was beneficial for soluble β-lactamase expression. (iii) The closer to the rbs the selected restriction site was, the more difficult it was to express soluble β-lactamase. (iv) The short-chain recombinant protein and the protein with His-tag fused at its C-terminus showed high affinity to the Ni2+ column. Conclusions Based on our findings, researchers can easily design an effective program for the high production of soluble recombinant β-lactamases to facilitate other related studies.

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“…The pH was determined by direct measurement in a digital potentiometer. The measurement of total soluble solids—TSS (°Brix) was performed using a digital refractometer (Quimis, Q767BD, Diadema, Brazil) and the concentration of lactic acid and acetic acid was determined by high-performance liquid chromatography (HPLC, Shimadzu Corp., Kyoto, Japan) [ 25 ].…”

Section: Methodsmentioning

confidence: 99%

“…Q767BD, Diadema, Brazil) and the concentration of lactic acid and acetic acid was determined by high-performance liquid chromatography (HPLC, Shimadzu Corp., Kyoto, Japan) [25]. Total protein concentration was determined by the Bradford method [26] using bovine serum albumin (BSA) as standard (Sigma Aldrich Co., St. Louis, MO, USA).…”

Section: Plos Onementioning

confidence: 99%

Fermented yellow mombin juice using Lactobacillus acidophilus NRRL B-4495: Chemical composition, bioactive properties and survival in simulated gastrointestinal conditions

et al. 2020

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The purpose of the present study was to evaluate yellow mombin (Spondias mombin L.) juice as a vehicle for the Lactobacillus acidophilus NRRL B-4495 probiotic. The initial pH and fermentation temperature conditions were optimized by central composite rotational design. The beverage was evaluated for its chemical composition, bioactive properties, microbiological stability, survival in simulated gastrointestinal conditions and sensory analysis. The ideal conditions for probiotic juice production were an initial pH of 6.4 and 16 h of fermentation, with maximum viability of 12.9 ± 0.4 Log CFU/mL. The fermented juice showed a total phenolic concentration of 94.90 ± 7.12 GAE/mL and antioxidant activity, as measured by DPPH (0.31 ± 0.00 μmol TE/mL) and ABTS sequestration (2.59 ± 0.30 μmol TE/mL). Antibacterial activity could also be observed against S. aureus, E. coli and K. pneumoniae. The obtained formulation showed good microbiological stability when stored at 4ºC for 28 days. In addition, there was no significant change in viability after exposure to simulated gastrointestinal conditions. The sensory analysis showed that the probiotic beverage was not well accepted. However, the Just-About-Right (JAR) ideal scale test enabled identifying the specific attributes which need to be improved from the tasters' point of view so that it is possible to improve product acceptance.

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Evaluation of induction conditions for plasmid pQE-30 stability and 503 antigen of Leishmania i. chagasi expression in E. coli M15

Cited by 11 publications

References 38 publications

Engineering a Rhamnose-Inducible System to Enhance the Extracellular Electron Transfer Ability of Shewanella Genus for Improved Cr(VI) Reduction

Engineering a Rhamnose-Inducible System to Enhance the Extracellular Electron Transfer Ability of Shewanella Genus for Improved Cr(VI) Reduction

Expression and purification of soluble recombinant β-lactamases using Escherichia coli as expression host and pET-28a as cloning vector

Fermented yellow mombin juice using Lactobacillus acidophilus NRRL B-4495: Chemical composition, bioactive properties and survival in simulated gastrointestinal conditions

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