recombinant escherichia coli
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Molecules ◽  
2022 ◽  
Vol 27 (1) ◽  
pp. 294
Fangting Wu ◽  
Ying Zhou ◽  
Wenyu Pei ◽  
Yuhan Jiang ◽  
Xiaohui Yan ◽  

Poly-(3-hydroxybutyrate) (PHB) is a polyester with biodegradable and biocompatible characteristics and has many potential applications. To reduce the raw material costs and microbial energy consumption during PHB production, cheaper carbon sources such as sucrose were evaluated for the synthesis of PHB under anaerobic conditions. In this study, metabolic network analysis was conducted to construct an optimized pathway for PHB production using sucrose as the sole carbon source and to guide the gene knockout to reduce the generation of mixed acid byproducts. The plasmid pMCS-sacC was constructed to utilize sucrose as a sole carbon source, and the cascaded promoter P3nirB was used to enhance PHB synthesis under anaerobic conditions. The mixed acid fermentation pathway was knocked out in Escherichia coli S17-1 to reduce the synthesis of byproducts. As a result, PHB yield was improved to 80% in 6.21 g/L cell dry weight by the resulted recombinant Escherichia coli in a 5 L bed fermentation, using sucrose as the sole carbon source under anaerobic conditions. As a result, the production costs of PHB will be significantly reduced.

Nivedhitha Swaminathan ◽  
Priyanka Priyanka ◽  
Anurag S. Rathore ◽  
Senthilkumar Sivaparakasam ◽  
Senthilmurugan Subbiah

2021 ◽  
pp. 105-115
Clóvis Moreira ◽  
Gustavo Marçal Schmidt Garcia Moreira ◽  
Rafael Amaral Donassolo ◽  
Marcos Roberto Alves Ferreira ◽  
Mariliana Luiza Ferreira Alves ◽  

Algorithms ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 326
Rafael Akira Akisue ◽  
Matheus Lopes Harth ◽  
Antonio Carlos Luperni Horta ◽  
Ruy de Sousa Junior

Due to low oxygen solubility and mechanical stirring limitations of a bioreactor, ensuring an adequate oxygen supply during a recombinant Escherichia coli cultivation is a major challenge in process control. Under the light of this fact, a fuzzy dissolved oxygen controller was developed, taking into account a decision tree algorithm presented in the literature, and implemented in the supervision software SUPERSYS_HCDC. The algorithm was coded in MATLAB with its membership function parameters determined using an Adaptive Network-Based Fuzzy Inference System tool. The controller was composed of three independent fuzzy inference systems: Princ1 and Princ2 assessed whether there would be an increment or a reduction in air and oxygen flow rates (respectively), whilst Delta estimated the size of these variations. To test the controller, simulations with a neural network model and E. coli cultivations were conducted. The fuzzification of the decision tree was successful, resulting in smoothing of air and oxygen flow rates and, hence, in an attenuation of dissolved oxygen oscillations. Statistically, the average standard deviation of the fuzzy controller was 2.45 times lower than the decision tree (9.48%). Results point toward an increase in the flow meter lifespan and a possible reduction of the metabolic stress suffered by E. coli during the cultivation.

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 357
Guey-Horng Wang ◽  
Chiu-Yu Cheng ◽  
Teh-Hua Tsai ◽  
Pin-Kuan Chiang ◽  
Ying-Chien Chung

In this study, we constructed a recombinant Escherichia coli strain with different promoters inserted between the chromate-sensing regulator chrB and the reporter gene luxAB to sense low hexavalent chromium (Cr(VI)) concentrations (<0.05 mg/L); subsequently, its biosensor characteristics (sensitivity, selectivity, and specificity) for measuring Cr(VI) in various water bodies were evaluated. The luminescence intensity of each biosensor depended on pH, temperature, detection time, coexisting carbon source, coexisting ion, Cr(VI) oxyanion form, Cr(VI) concentration, cell type, and type of medium. Recombinant lux-expressing E. coli with the T7 promoter (T7-lux-E. coli, limit of detection (LOD) = 0.0005 mg/L) had the highest luminescence intensity or was the most sensitive for Cr(VI) detection, followed by E. coli with the T3 promoter (T3-lux-E. coli, LOD = 0.001 mg/L) and that with the SP6 promoter (SP6-lux-E. coli, LOD = 0.005 mg/L). All biosensors could be used to determine whether the Cr(VI) standard was met in terms of water quality, even when using thawing frozen cells as biosensors after 90-day cryogenic storage. The SP6-lux-E. coli biosensor had the shortest detection time (0.5 h) and the highest adaptability to environmental interference. The T7-lux-E. coli biosensor—with the optimal LOD, a wide measurement range (0.0005–0.5 mg/L), and low deviation (−5.0–7.9%) in detecting Cr(VI) from industrial effluents, domestic effluents, and surface water—is an efficient Cr(VI) biosensor. This unprecedented study is to evaluate recombinant lux E. coli with dissimilar promoters for their possible practice in Cr(VI) measurement in water bodies, and the biosensor performance is clearly superior to that of past systems in terms of detection time, LOD, and detection deviation for real water samples.

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