Enhanced production of nonanedioic acid from nonanoic acid by engineered <i>Escherichia coli</i>

Lee, Yong-Joo; Sathesh‐Prabu, Chandran; Kwak, Geun Hwa; Bang, Ina; Jung, Hyun Wook; Kim, Donghyuk; Lee, Sung Kuk

doi:10.1002/biot.202000416

Cited by 9 publications

(2 citation statements)

References 57 publications

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“…In order to study the applicability of the fusion enzyme catalysis in bioremediation we performed a reaction scale-up exploiting a strain of E. coli BL21 (DE3) cells transformed with the pET-28-a-CYP116B5-SOX(+) vector: E. coli (CYP116B5-SOX). We employed the transformed bacteria as biocatalytic system to remove tamoxifen, [64][65][66][67][68][69][70][71][72][73] commonly reported as water pollutant, [9] from an aqueous buffered medium, taken as a model of contaminated water. We used HPLC-MS to identify the metabolites produced by CYP116B5-SOX catalysis.…”

Section: Drugs Bioconversion Using Cyp 116b5-sox Expressing E Coli St...mentioning

confidence: 99%

CYP116B5‐SOX: An artificial peroxygenase for drug metabolites production and bioremediation

Giuriato,

Catucci,

Correddu

et al. 2024

Biotechnology Journal

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CYP116B5 is a class VII P450 in which the heme domain is linked to a FMN and 2Fe2S‐binding reductase. Our laboratory has proved that the CYP116B5 heme domain (CYP116B5‐hd) is capable of catalyzing the oxidation of substrates using H2O2. Recently, the Molecular Lego approach was applied to join the heme domain of CYP116B5 to sarcosine oxidase (SOX), which provides H2O2 in‐situ by the sarcosine oxidation. In this work, the chimeric self‐sufficient fusion enzyme CYP116B5‐SOX was heterologously expressed, purified, and characterized for its functionality by absorbance and fluorescence spectroscopy. Differential scanning calorimetry (DSC) experiments revealed a TM of 48.4 ± 0.04 and 58.3 ± 0.02°C and a enthalpy value of 175,500 ± 1850 and 120,500 ± 1350 cal mol−1 for the CYP116B5 and SOX domains respectively. The fusion enzyme showed an outstanding chemical stability in presence of up to 200 mM sarcosine or 5 mM H2O2 (4.4 ± 0.8 and 11.0 ± 2.6% heme leakage respectively). Thanks to the in‐situ H2O2 generation, an improved kcat/KM for the p‐nitrophenol conversion was observed (kcat of 20.1 ± 0.6 min−1 and KM of 0.23 ± 0.03 mM), corresponding to 4 times the kcat/KM of the CYP116B5‐hd. The aim of this work is the development of an engineered biocatalyst to be exploited in bioremediation. In order to tackle this challenge, an E. coli strain expressing CYP116B5‐SOX was employed to exploit this biocatalyst for the oxidation of the wastewater contaminating‐drug tamoxifen. Data show a 12‐fold increase in tamoxifen N‐oxide production—herein detected for the first time as CYP116B5 metabolite—compared to the direct H2O2 supply, equal to the 25% of the total drug conversion.

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Section: Drugs Bioconversion Using Cyp 116b5-sox Expressing E Coli St...mentioning

confidence: 99%

CYP116B5‐SOX: An artificial peroxygenase for drug metabolites production and bioremediation

Giuriato,

Catucci,

Correddu

et al. 2024

Biotechnology Journal

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“…Compared with metabolic engineering, ALE allows the redirection of metabolism without considering the metabolic networks of the microbial cells. It has recently been widely applied for improving carbon source utilization 23 , 24 , enhancing the production efficiency of target biomolecules 25 – 27 , and creating stress-tolerant microorganisms toward thermal, ethanol, acetic acid, and lignocellulosic inhibitors 13 , 28 – 30 . Regarding the application of ALE for the improvement of yeast properties, most studies have focused on the conventional species S. cerevisiae 31 – 37 .…”

Section: Introductionmentioning

confidence: 99%

Adaptive laboratory evolution under acetic acid stress enhances the multistress tolerance and ethanol production efficiency of Pichia kudriavzevii from lignocellulosic biomass

Dolpatcha,

Phong,

Thanonkeo

et al. 2023

Sci Rep

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Second-generation bioethanol production using lignocellulosic biomass as feedstock requires a highly efficient multistress-tolerant yeast. This study aimed to develop a robust yeast strain of P. kudriavzevii via the adaptive laboratory evolution (ALE) technique. The parental strain of P. kudriavzevii was subjected to repetitive long-term cultivation in medium supplemented with a gradually increasing concentration of acetic acid, the major weak acid liberated during the lignocellulosic pretreatment process. Three evolved P. kudriavzevii strains, namely, PkAC-7, PkAC-8, and PkAC-9, obtained in this study exhibited significantly higher resistance toward multiple stressors, including heat, ethanol, osmotic stress, acetic acid, formic acid, furfural, 5-(hydroxymethyl) furfural (5-HMF), and vanillin. The fermentation efficiency of the evolved strains was also improved, yielding a higher ethanol concentration, productivity, and yield than the parental strain, using undetoxified sugarcane bagasse hydrolysate as feedstock. These findings provide evidence that ALE is a practical approach for increasing the multistress tolerance of P. kudriavzevii for stable and efficient second-generation bioethanol production from lignocellulosic biomass.

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Biosynthesis of Odd-Carbon Unsaturated Fatty Dicarboxylic Acids Through Engineering the HSAF Biosynthetic Gene in Lysobacter enzymogenes

Khetrapal

Dussault

2022

Mol Biotechnol

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Enhanced production of nonanedioic acid from nonanoic acid by engineered Escherichia coli

Cited by 9 publications

References 57 publications

CYP116B5‐SOX: An artificial peroxygenase for drug metabolites production and bioremediation

CYP116B5‐SOX: An artificial peroxygenase for drug metabolites production and bioremediation

Adaptive laboratory evolution under acetic acid stress enhances the multistress tolerance and ethanol production efficiency of Pichia kudriavzevii from lignocellulosic biomass

Biosynthesis of Odd-Carbon Unsaturated Fatty Dicarboxylic Acids Through Engineering the HSAF Biosynthetic Gene in Lysobacter enzymogenes

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