Expression of genes encoding the luciferase from <i>Photobacterium leiognathi</i> in <scp><i>Escherichia coli</i></scp> Rosetta (DE3) and its application in NADH detection

Xuan, Guanhua; Xiao, Qilin; Wang, Jingxue; Lin, Hong; Pavase, Tushar Ramesh

doi:10.1002/bio.3501

Cited by 8 publications

(4 citation statements)

References 48 publications

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“…In this system, crude lysate of Lux from Photobacterium leiognathi YL expressing Escherichia coli cells containing an endogenous Fre was used. [ 69 ] Later on, the detection limit of this system could be enhanced with a detection range of 0.1–1 n m when the purified recombinant LuxG and Lux enzymes were used instead. [ 70 ] LuxG, a flavin reductase found in the same operon as Lux, prefers to use FMN more than other flavins, while Fre has more preference toward riboflavin.…”

Section: Nad(p)h Detection As An Indicator For Energy Level and Cell ...mentioning

confidence: 99%

Detection of cellular metabolites by redox enzymatic cascades

Tinikul

Trisrivirat

Chinantuya

et al. 2022

Biotechnology Journal

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Detection of cellular metabolites that are disease biomarkers is important for human healthcare monitoring and assessing prognosis and therapeutic response. Accurate and rapid detection of microbial metabolites and pathway intermediates is also crucial for the process optimization required for development of bioconversion methods using metabolically engineered cells. Various redox enzymes can generate electrons that can be employed in enzyme‐based biosensors and in the detection of cellular metabolites. These reactions can directly transform target compounds into various readout signals. By incorporating engineered enzymes into enzymatic cascades, the readout signals can be improved in terms of accuracy and sensitivity. This review critically discusses selected redox enzymatic and chemoenzymatic cascades currently employed for detection of human‐ and microbe‐related cellular metabolites including, amino acids, d‐glucose, inorganic ions (pyrophosphate, phosphate, and sulfate), nitro‐ and halogenated phenols, NAD(P)H, fatty acids, fatty aldehyde, alkane, short chain acids, and cellular metabolites.

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Section: Nad(p)h Detection As An Indicator For Energy Level and Cell ...mentioning

confidence: 99%

Detection of cellular metabolites by redox enzymatic cascades

Tinikul

Trisrivirat

Chinantuya

et al. 2022

Biotechnology Journal

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“…SDS-PAGE was carried out to evaluate the possible disruption of bacterial protein expression induced by OTA exposure (Xuan et al, 2018). The bacterial pellet of treatment group was exposed to 40 mg L À1 OTA solution for 30 min, and sterile saline severed as a blank control.…”

Section: Sds-page For Proteins Assaymentioning

confidence: 99%

Rapid detection and toxicity assessment of ochratoxin A by Photobacterium leiognathi in drinking water

Yang

et al. 2019

Int J of Food Sci Tech

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Summary This study was aimed to develop a rapid technique for detection of ochratoxin A (OTA) by Photobacterium leiognathi (P. leiognathi) based on its inhibition of luminescence on P. leiognathi. The freeze‐dried powder of P. leiognathi was incubated and grown aerobically in sterile liquid medium at 28 °C for 20 h. Optical cell density (OD) at wavelength of 610 nm was measured using UV spectrometer every 2 h. Different concentrations of standard OTA solution were used to measure its luminescence inhibition and calculate its half maximal inhibitory concentration (IC50). Scanning electron microscopy (SEM), flow cytometry (FCM), sodium dodecyl sulphonate polyacrylamide gel electrophoresis (SDS‐PAGE), DNA extraction and gel electrophoresis were used to observe the performance of P. leiognathi under the treatment of OTA. A correlation (R2 > 0.98) was obtained between the relative luminosity unit of P. leiognathi and OTA concentration in the range of 0.01–20 mg L−1 with recoveries of 80.8–87.4%. The effects of OTA on P. leiognathi are time‐dependent, and the IC50 value of 12.71 mg L−1 at 30 min demonstrated its good sensitivity to OTA. The cells of P. leiognathi under 40 mg L−1 OTA exposure for 30 mins showed morphological alterations, protein damage, apoptosis and necrosis. The aforementioned results indicate that biological assay is a promising and alternative method used for rapidly monitoring the sudden pollution of OTA in the early emergency warning of drinking water system.

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“…Herein, a novel lipase M16 derived from Aspergillus oryzae was cloned and expressed, and its enzymatic properties were characterized. A new recombinant engineered bacterium expressing the Aspergillus oryzae lipase gene was constructed as a biocatalyst using E. coli Rosetta (DE3) as an expression host; it was found suitable for the expression of fungal‐derived genes 21,22 . The reaction factors as temperature, pH, and cosolvent were investigated for the enantioselective resolution of the substrate ( R , S )‐methyl 2‐propylsuccinate 4‐ tert ‐butyl ester to synthesize the BRV intermediate, ( R )‐2‐propylsuccinic acid 4‐ tert ‐butyl ester.…”

Section: Introductionmentioning

confidence: 99%

A novel lipase from Aspergillus oryzae WZ007 catalyzed synthesis of brivaracetam intermediate and its enzymatic characterization

Zhang

et al. 2020

Chirality

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Brivaracetam is a structural derivative of the chiral drug levetiracetam and has been approved for the adjuvant treatment of partial epilepsy. As a new antiepileptic drug, it is widely used in a variety of epilepsy models. In this study, a novel lipase M16 derived from Aspergillus oryzae WZ007 was cloned, expressed, and used for chiral resolution. Lipase M16 has a high enantioselectivity to the racemic substrate (R,S)-methyl 2-propylsuccinate 4-tert-butyl ester, and the intermediate (R)-2-propylsuccinic acid 4-tert-butyl ester of brivaracetam was obtained efficiently. Under optimal conditions, the enantiomeric excess of substrate was up to 99.26%, and the e.e. p was 96.23%. The conversion and apparent E value were 50.63% and 342.48, respectively. This study suggests a new biocatalytic resolution via lipase M16 for preparing the brivaracetam chiral intermediate and its potential application in the pharmaceutical industry.

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Expression of genes encoding the luciferase from Photobacterium leiognathi in Escherichia coli Rosetta (DE3) and its application in NADH detection

Cited by 8 publications

References 48 publications

Detection of cellular metabolites by redox enzymatic cascades

Detection of cellular metabolites by redox enzymatic cascades

Rapid detection and toxicity assessment of ochratoxin A by Photobacterium leiognathi in drinking water

A novel lipase from Aspergillus oryzae WZ007 catalyzed synthesis of brivaracetam intermediate and its enzymatic characterization

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