Biocatalytic conversion of ethylene to ethylene oxide using an engineered toluene monooxygenase

A high-throughput cell analysis and sorting platform using droplet-based microfluidics is introduced for directed evolution of recombinant CotA laccase expressed in E. coli.

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Over-expression of human DNA polymerase lambda in E. coli and characterization of the recombinant enzyme

Genes to Cells ◽

10.1046/j.1365-2443.2002.00547.x ◽

2002 ◽

Vol 7 (7) ◽

pp. 639-651 ◽

Cited By ~ 88

Author(s):

Noriko Shimazaki ◽

Kenta Yoshida ◽

Toshiko Kobayashi ◽

Shingo Toji ◽

Katsuyuki Tamai ◽

...

Keyword(s):

Dna Polymerase ◽

Recombinant Enzyme ◽

Over Expression ◽

E Coli ◽

Human Dna

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Mutagenic effects of ethylene oxide in E. coli

Mutation Research/Environmental Mutagenesis and Related Subjects ◽

10.1016/0165-1161(88)90136-7 ◽

1988 ◽

Vol 203 (3) ◽

pp. 215-216

Author(s):

A. Kolman ◽

M. Näslund

Keyword(s):

Ethylene Oxide ◽

E Coli ◽

Mutagenic Effects

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Protective effect of low doses in mutagenesis with ethylene oxide in E. coli

Mutation Research Letters ◽

10.1016/0165-7992(84)90122-2 ◽

1984 ◽

Vol 139 (4) ◽

pp. 167-171

Author(s):

Ada Kolman

Keyword(s):

Ethylene Oxide ◽

Protective Effect ◽

Low Doses ◽

E Coli

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Fabrication and Super-Antibacterial Property of Nanosilver/Sericin/Poly(ethylene oxide) Nanofibers through Electrospinning-Combined Postdeposition Method

Journal of Nanomaterials ◽

10.1155/2016/9859530 ◽

2016 ◽

Vol 2016 ◽

pp. 1-7 ◽

Cited By ~ 2

Author(s):

Jia Li ◽

Bo-Xiang Wang ◽

Yi-Fan Cui ◽

Zhi-Cai Yu ◽

Xu Hao ◽

...

Keyword(s):

Escherichia Coli ◽

Ethylene Oxide ◽

Antibacterial Property ◽

Silver Content ◽

Antibacterial Properties ◽

Bacterial Reduction ◽

E Coli ◽

Poly Ethylene Oxide ◽

Poly Ethylene ◽

Nanofiber Mats

Nanosilver particle has been used in the nanofiber mats by mixing the nanosilver with the spinning solution for improving the antibacterial property. Although studies have shown that the antibacterial property of nanofiber mats gets increasing, the higher silver content and the larger released resistance of nanosilver from nanofiber mats are obvious. Here, the electrospinning-combined postdeposition method was used to prepare the nanosilver/sericin/poly(ethylene oxide) (Ag/SS/PEO) nanofiber mats and the bacterial reduction rates againstStaphylococcus aureus(S. aureus) andEscherichia coli(E. coli) were analyzed. We found that the Ag/SS/PEO nanofiber mats were excellent antibacterial properties at the lower silver content and the bacterial reduction rates againstS. aureusandE. coliall reached above 99.99%. Our data suggests that the antibacterial property can be improved by introducing the electrospinning-combined postdeposition method.

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Rat Muscle Fructose-1,6-Bisphosphatase: Cloning of the cDNA, Expression of the Recombinant Enzyme, and Expression Analysis in Different Tissues

Biological Chemistry ◽

10.1515/bc.1999.134 ◽

1999 ◽

Vol 380 (9) ◽

pp. 1079-1085 ◽

Cited By ~ 27

Author(s):

Samiya Al-Robaiy ◽

Klaus Eschrich

Keyword(s):

Northern Blotting ◽

Recombinant Enzyme ◽

Rat Tissues ◽

Mrna Levels ◽

Coding Region ◽

Total Rna ◽

E Coli ◽

Rat Muscle ◽

Positional Identity ◽

Fructose 1,6 Bisphosphatase

Abstract The 1282 bp cDNA of an isoenzyme of fructose-1,6- bisphosphatase was cloned from rat muscle. It shows 70% positional identity to the cDNA of rat liver fructose- 1,6-bisphosphatase and is clearly the product of a gene different from that coding for the liver enzyme. After cloning of the coding region of the rat muscle fructose-1,6-bisphosphatase cDNA in an expression vector, the recombinant enzyme could be detected in E. coli cell-free extracts by activity determination and Western blotting. Overexpressed fructose-1,6-bisphosphatase was found to be allosterically inhibited by AMP comparably to the enzyme isolated from rat muscle. Analysis of steady-state mRNA levels of various rat tissues with reverse-transcriptase polymerase chain reaction (RT-PCR) and Northern blotting revealed one or the two fructose-1,6-bisphosphatase isoenzyme mRNAs in most tissues tested with significant quantitative differences. Quantitative PCR using a homologous competitor showed that 1 μg of total RNA of rat muscle contains 1.7 × 106 molecules of rat muscle fructose-1,6-bisphosphatase mRNA. 3 × 104 copies of this message were found per μg total RNA of heart and kidney, respectively.

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Heterologous Expression and Application of Multicopper Oxidases from Enterococcus spp. for Degradation of Biogenic Amines

Protein and Peptide Letters ◽

10.2174/0929866527666200616160859 ◽

2020 ◽

Vol 27 ◽

Author(s):

Binbin Li ◽

Yuan Wang ◽

Linlin Xue ◽

Shiling Lu

Keyword(s):

Gene Expression ◽

Biogenic Amines ◽

Multicopper Oxidase ◽

Recombinant Enzyme ◽

Alkali Resistance ◽

Prolonged Incubation ◽

E Coli ◽

Recombinant Strains ◽

Enterococcus Spp ◽

Amine Degradation

Background: Biogenic amines are harmful to human health at a certain extent. As a kind of biogenic amine oxidase, multicopper oxidase can be used to degrade them. Currently, the literature about enzyme from Enterococcus spp. are limited, and recombinant multicopper oxidase might be an effective way to degrade biogenic amines. Objective: (i) Select and identify strains that can degrade biogenic amines, (ii) overexpress enzyme from Enterococcus spp., (iii) measure gene expression and probe amine-degradation differences among strains (native, E. coli DH5α, and L. delbruckii), and (iv) examine the biochemical properties of recombinant multicopper oxidase, (v) apply the recombinant enzyme into smoked horsemeat sausage. Methods: Reverse transcription PCR and high-performance liquid chromatography were performed to examine gene expression and amine degradation rate. Results: The results demonstrated that target enzymes were successfully overexpressed, accompanied by increased aminedegrading activity (P <0.05). Gene from E. faecalis M5B was expressed in L. delbrueckii resulted in degradation rates for phenylethylamine, putrescine, histamine and tyramine of 54%, 52%, 70% and 40%, respectively, significantly higher than achieved by other recombinant strains. Conclusion: In this work, gene expression levels were higher in recombinant M5B than recombinant M2B, regardless of host. E. coli is more stable to express multicopper oxidase. Besides, the amine-degrading ability was markedly increased in the two recombinant strains. After prolonged incubation, the recombinant enzyme could degrade three amines, and it displayed high alkali resistance and thermostability.

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Preparation of chitosan-containing nanofibres by electrospinning of chitosan/poly(ethylene oxide) blend solutions

e-Polymers ◽

10.1515/epoly.2004.4.1.624 ◽

2004 ◽

Vol 4 (1) ◽

Cited By ~ 38

Author(s):

Maria Spasova ◽

Nevena Manolova ◽

Dilyana Paneva ◽

Iliya Rashkov

Keyword(s):

Field Strength ◽

Ethylene Oxide ◽

Applied Field ◽

Total Concentration ◽

E Coli ◽

Antimycotic Activity ◽

Poly Ethylene Oxide ◽

Antimycotic Agent ◽

Model Drug ◽

Poly Ethylene

Abstract The first successful preparation of chitosan-containing nanofibres was achieved by electrospinning of chitosan/poly(ethylene oxide) (PEO) blend aqueous solutions. The diameters of the nanofibres were in the range 40 - 290 nm and decreased with increasing chitosan content and decreasing total concentration. An increase of the applied field strength leads to an increase of the diameter of the nanofibres and to a broadening of the size distribution. The possibility to prepare nanofibres containing a model drug - potassium 5-nitro-8-quinolinolate (K5N8Q), a broad-spectrum antimicrobial and antimycotic agent - was shown. The incorporation of K5N8Q in the nanofibres resulted in a decrease of the nanofibre diameters and the appearance of bead-shaped defects. Non-woven mats from the drugloaded nanofibres with composition chitosan : PEO = 1:1 (w/w) and 1% K5N8Q showed antibacterial and antimycotic activity against E. coli, S. aureus and C.albicans.

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Cloning and Expression of Phytase appA Gene from Shigella sp. CD2 in Pichia pastoris and Comparison of Properties with Recombinant Enzyme Expressed in E. coli

PLoS ONE ◽

10.1371/journal.pone.0145745 ◽

2016 ◽

Vol 11 (1) ◽

pp. e0145745 ◽

Cited By ~ 8

Author(s):

Moushree Pal Roy ◽

Deepika Mazumdar ◽

Subhabrata Dutta ◽

Shyama Prasad Saha ◽

Shilpi Ghosh

Keyword(s):

Pichia Pastoris ◽

Recombinant Enzyme ◽

Cloning And Expression ◽

E Coli

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Optimization of Fermentation Conditions for Recombinant Lipase Expression At Small Scale Using Response Surface Methodology for Preliminary Production In Two Bioreactor Platforms.

10.21203/rs.3.rs-152763/v1 ◽

2021 ◽

Author(s):

Angela Liliana Meza López ◽

Alejandro Acosta-González ◽

Ingrid Yamile Pulido Manrique ◽

Rosa Erlide Prieto Correa ◽

Carlos Jimenez Junca

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Lipase Activity ◽

Input Parameter ◽

Power Input ◽

Recombinant Enzyme ◽

Fermentation Conditions ◽

E Coli ◽

Recombinant Lipase ◽

Lipase Expression

Abstract Background: Pseudomonas lipases are widely used in industrial applications due to its unique biochemical properties, but one of the biggest limitations are the low yields obtained in native strains therefore, organisms as E. coli are used for the recombinant lipase overexpression. However, the recombinant lipase is accumulated as inclusion bodies and it affects biological activity, making that researchers evaluate different fermentation conditions to improve the activity of recombinant enzymes. In this study, a statistical experimental design was implemented to evaluate the effects of temperature, agitation rate and osmolyte concentration on the recombinant lipase activity produced in E. coli BL21 (DE3). Once the significant variables were identified, an optimization by a Response Surface Methodology was applied to maximize the lipase production. Results: The Box-Behnken designs revealed different optimal fermentation conditions for each osmolyte experiment. The glycerol showed the highest specific lipase activity compared to the other osmolytes and 0.1 M of osmolyte glycerol,5°C and 110 rpm showed the highest significant increase on the specific lipase activity and the data fitted the model very well. The validation showed that 452.01 U/mg of specific lipase activity was obtained which was significantly higher compared to the group where no glycerol was added (271.38 U/mg). The relative recombinant lipase expression was 2.7-fold lower at 5°C compared to 25 °C, but at 5°C the lipase activity was significantly higher. In addition, when the 3 L shaken Erlenmeyer Bioreactor was used to produce the recombinant lipase based on the power input parameter, the specific lipase activity was not significantly different from that found in Schott (408,4 U/mg and 452 U/mg, respectively), which means that this Bioreactor platform should be used for future scale-up processes.Conclusion: Low temperatures, low agitation rates and 0.1 M of glycerol in the autoinduction media enhanced the activity of the recombinant lipase produced in E. coli BL21(DE3). The optimized conditions and the 3 L shaken Erlenmeyer Bioreactor can be used to produce the recombinant enzyme in a higher volume based on the power input parameter. Further studies using this strategy may lead to the identification of optimal culture conditions for a given recombinant enzyme facilitating the large-scale bioprocess implementation.

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