Endogenous microenvironmental engineering through targeted alteration of salt bridge network can effectively regulate enzymatic pH adaptation

Li, Zonglin; Sun, Chuanqi; Lou, Longwei; Li, Zhimin

doi:10.1016/j.cej.2022.136215

Cited by 9 publications

(3 citation statements)

References 48 publications

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“…Additionally, CarEW has a broader pH range (6.5–9.5) than pnbA (6–8). The presence of more thermostabilizing residues may conversely narrow the pH optimum of an enzyme [53] . It is suggested other research groups consider this as a potential tradeoff.…”

Section: Resultsmentioning

confidence: 99%

Identification and characterisation of moderately thermostable diisobutyl phthalate degrading esterase from a Great Artesian Basin Bacillus velezensis NP05

Mu,

Sadowski,

Te'o

et al. 2024

Biotechnology Reports

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Section: Resultsmentioning

confidence: 99%

Identification and characterisation of moderately thermostable diisobutyl phthalate degrading esterase from a Great Artesian Basin Bacillus velezensis NP05

Mu,

Sadowski,

Te'o

et al. 2024

Biotechnology Reports

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“…The cultivation conditions and protein purification steps were modified with those established by our previous studies (Li et al, 2020; Li et al, 2022). E. coli BL21(DE3) which was engineered for expression of DecR variants was cultivated in LB medium containing 100 μg/ml ampicillin for 8 h, and then 3 ml preincubate culture was inoculated in 200 ml fresh LB medium and then cultivated overnight at 25°C.…”

Section: Methodsmentioning

confidence: 99%

Engineering of the Lrp/AsnC‐type transcriptional regulator DecR as a genetically encoded biosensor for multilevel optimization of L‐cysteine biosynthesis pathway in Escherichia coli

Zhou,

Li,

Zhong

et al. 2024

Biotech & Bioengineering

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L‐cysteine is an important sulfur‐containing amino acid being difficult to produce by microbial fermentation. Due to the lack of high‐throughput screening methods, existing genetically engineered bacteria have been developed by simply optimizing the expression of L‐cysteine‐related genes one by one. To overcome this limitation, in this study, a biosensor‐based approach for multilevel biosynthetic pathway optimization of L‐cysteine from the DecR regulator variant of Escherichia coli was applied. Through protein engineering, we obtained the DecRN29Y/C81E/M90Q/M99E variant‐based biosensor with improved specificity and an 8.71‐fold increase in dynamic range. Using the developed biosensor, we performed high‐throughput screening of the constructed promoter and RBS combination library, and successfully obtained the optimized strain, which resulted in a 6.29‐fold increase in L‐cysteine production. Molecular dynamics (MD) simulations and electrophoretic mobility shift analysis (EMSA) showed that the N29Y/C81E/M90Q/M99E variant had enhanced induction activity. This enhancement may be due to the increased binding of the variant to DNA in the presence of L‐cysteine, which enhances transcriptional activation. Overall, our biosensor‐based strategy provides a promising approach for optimizing biosynthetic pathways at multiple levels. The successful implementation of this strategy demonstrates its potential for screening improved recombinant strains.

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“…The invention of the pH glass electrode revolutionized pH measurement, providing an accurate and reliable method compared to traditional chemical indicators and pH measurement gradually became essential to control and optimize reactions involved in biological processes, environmental monitoring, and industrial processes. Indeed, pH regulation is essential for enzymatic function and physiological balance [5,6], while pH monitoring makes it possible to control industrial processes to ensure the safety of products in the agri-food sector [7,8]. In addition, pH measurement makes it possible to better manage agricultural soils [9,10], but also to contribute to environmental monitoring through the assessment of water quality or ecosystem health [11,12].…”

Section: Introductionmentioning

confidence: 99%

Polyaniline-Based Flexible Sensor for pH Monitoring in Oxidizing Environments

Bignall,

Magnenet,

Ramsamy

et al. 2024

Chemosensors

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Measuring pH in oxidizing solutions is a crucial issue in areas such as aquaculture, water treatment, industrial chemistry, and environmental analysis. For this purpose, a low-cost potentiometric flexible sensor using a polymer film as a pH-sensitive material has been developed in this study. The sensor consists in a polyaniline film electrodeposited from a sulfuric acid solution on a gold electrode previously deposited on a flexible polyimide substrate. The resulting polyaniline-based pH sensors showed an interesting performance detection in aqueous solution, leading to sensitive (73.4 mV per unit pH) and reproducible (standard deviation of 1.75) responses over the entire pH range from 3 to 8. On the contrary, they were inoperative in the presence of oxidizing hypochlorite ions. Thus, other polyaniline films were electrodeposited in the presence of cetyltrimethylammonium bromide or Tritonx100 surfactant in an attempt to improve the sensing performance of the pH sensors in oxidizing solutions. The pH sensors based on polyaniline and Tritonx100 surfactant were then found to be sensitive (62.3 mV per unit pH) and reproducible (standard deviation of 1.52) in aqueous solutions containing hypochlorite ions. All polyaniline films were also characterized by profilometry and electronic microscopy to correlate the physicochemical features with the performance of the sensors.

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Endogenous microenvironmental engineering through targeted alteration of salt bridge network can effectively regulate enzymatic pH adaptation

Cited by 9 publications

References 48 publications

Identification and characterisation of moderately thermostable diisobutyl phthalate degrading esterase from a Great Artesian Basin Bacillus velezensis NP05

Identification and characterisation of moderately thermostable diisobutyl phthalate degrading esterase from a Great Artesian Basin Bacillus velezensis NP05

Engineering of the Lrp/AsnC‐type transcriptional regulator DecR as a genetically encoded biosensor for multilevel optimization of L‐cysteine biosynthesis pathway in Escherichia coli

Polyaniline-Based Flexible Sensor for pH Monitoring in Oxidizing Environments

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