High-level soluble expression of bioactive porcine myostatin propeptide in E. coli

2013 ◽  
Vol 97 (19) ◽  
pp. 8517-8527 ◽  
Author(s):  
Wing Yeung Haq ◽  
Sang Kee Kang ◽  
Sang Beum Lee ◽  
Hee Chul Kang ◽  
Yun Jaie Choi ◽  
...  
Keyword(s):  
Soluble Expression ◽  
E Coli ◽  
High Level

High-level soluble expression of isopenicillin N synthase isozymes in E. coli

Tetrahedron ◽  
1991 ◽  
Vol 47 (31) ◽  
pp. 5991-6002 ◽  
Author(s):  
J.E. Baldwin ◽  
J.M. Blackburn ◽  
J.D. Sutherland ◽  
M.C. Wright
Keyword(s):  
Soluble Expression ◽  
E Coli ◽  
High Level ◽  
Isopenicillin N

scholarly journals High-level soluble expression and enzymatic characterization of Burkholderia sp. lipase with steryl ester hydrolysis activity in E. coli

2019 ◽  
Author(s):  
Zhengyu Shu ◽  
Feng Li ◽  
Wenjing Jia ◽  
Xiangduo Mu ◽  
Hong Lin ◽  
...  
Keyword(s):  
Steryl Ester ◽  
Specific Activity ◽  
Soluble Expression ◽  
Hydrolase Activity ◽  
Biodiesel Production ◽  
Enzymatic Characterization ◽  
E Coli ◽  
Ester Hydrolase ◽  
Triacylglycerol Hydrolase ◽  
High Level

Abstract Background Burkholderia cepacia lipase is an important industrial biocatalyst for biodiesel production and chiral pharmaceutical synthesis. Heterologous soluble expression of lipase lipA gene from B. cepacia in Escherichia coli highly depends on co-expression of its cognate foldase gene, lipB. However, the interaction between recombinant lipase LipA and chaperonin LipB is rather complicated and confusing. In this research, various systems of lipA/lipB co-expression combinations are investigated to obtain high-level soluble expression of lipA, respectively. Results The best co-expression combination system for lipA and lipB is E. coli Origami 2 (DE3)/pETDuet-lipB(MCS1)/lipA(MCS2). The soluble expression level of lipA is 100.4 U/OD600 towards 4-nitrophenyl laurate hydrolysis. The recombinant LipA can be rapidly isolated from cell-free supernatant of recombinant E. coli lysate using HisTrap HP affinity chromatography column, and the lipA/LipB complex is obtained. Enzymatic characterization analysis shows that the purified LipA is a mesothermal and alkaline enzyme. LipA displays preference for medium-chain-length acyl groups (C10-C12) and sn-1,3 regioselectivity. Besides triacylglycerol hydrolase activity (EC. 3.1.1.3), LipA also displays steryl ester hydrolase activity (EC. 3.1.1.13). The specific activity of LipA towards 4-nitrophenyl decanoate and cholesterol linoleate are 638.9 U/mg and 1111.5 mU/mg, respectively. Conclusions Host strain E. coli Origami 2 (DE3), lipB locus at MCS1 on the dual expression cassette plasmid pETDuet, and low-temperature induction contribute to the soluble expression of lipA. Recombinant LipA displays both triacylglycerol hydrolase activity and steryl ester hydrolase activity.

Toxic Effect of 2-ethyl (bicyclo[2.2.1] heptane) on Bacterial Cells

2019 ◽  
Vol 35 (6) ◽  
pp. 67-72 ◽  
Author(s):  
I.V. Manukhov ◽  
L.S. Yaguzhinsky ◽  
M.V. Bermeshev ◽  
M.A. Zisman ◽  
V.G. Pevgov ◽  
...  
Keyword(s):  
Toxic Effect ◽  
Atmospheric Oxygen ◽  
Inducible Promoter ◽  
Oxygen Species ◽  
Bacterial Cells ◽  
Unique Identifier ◽  
E Coli ◽  
Lux Biosensors ◽  
High Level ◽  
Ministry Of Higher Education

Toxic effect of 2-ethylnorbornane (2-ethyl(bicyclo[2.2.1]heptane) (EBH)) on bacteria has been studied using the E. coli pRecA-lux and E. coli pKatG- lux cells as lux-biosensors. It was shown that the addition of EBH to the incubation medium leads to death and growth retardation, high level oxidative stress and DNA damage in E. coli cells. It is assumed that the oxidation of EBH with atmospheric oxygen causes the formation of reactive oxygen species in the medium, which makes a major contribution to the toxicity of this substance. biosensor, luciferase, bioluminescence, inducible promoter, PrecA, PkatG The authors are grateful to Stanislav Filippovich Chalkin for the development of interdisciplinary ties in the scientific community. The work was financially supported by the Ministry of Higher Education and Science of Russia (Project Unique Identifier RFMEFI60417X0181, Agreement No. 14.604.21.0181 of 26.09.2017).

scholarly journals Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA

2021 ◽  
Vol 22 (15) ◽  
pp. 7843
Author(s):  
Sang-Oh Ahn ◽  
Ho-Dong Lim ◽  
Sung-Hwan You ◽  
Dae-Eun Cheong ◽  
Geun-Joong Kim
Keyword(s):  
Tertiary Structure ◽  
Signal Sequence ◽  
Soluble Expression ◽  
Class I ◽  
Soluble Form ◽  
Two Phase ◽  
E Coli ◽  
Small Proteins ◽  
Industrial Use ◽  
Shed Light

Hydrophobins are small proteins (<20 kDa) with an amphipathic tertiary structure that are secreted by various filamentous fungi. Their amphipathic properties provide surfactant-like activity, leading to the formation of robust amphipathic layers at hydrophilic–hydrophobic interfaces, which make them useful for a wide variety of industrial fields spanning protein immobilization to surface functionalization. However, the industrial use of recombinant hydrophobins has been hampered due to low yield from inclusion bodies owing to the complicated process, including an auxiliary refolding step. Herein, we report the soluble expression of a recombinant class I hydrophobin DewA originating from Aspergillus nidulans, and its efficient purification from recombinant Escherichia coli. Soluble expression of the recombinant hydrophobin DewA was achieved by a tagging strategy using a systematically designed expression tag (ramp tag) that was fused to the N-terminus of DewA lacking the innate signal sequence. Highly expressed recombinant hydrophobin DewA in a soluble form was efficiently purified by a modified aqueous two-phase separation technique using isopropyl alcohol. Our approach for expression and purification of the recombinant hydrophobin DewA in E. coli shed light on the industrial production of hydrophobins from prokaryotic hosts.

High-Level Expression of Human β-Defensin-2 Gene with Rare Codons in E. coli Cell-Free System

2006 ◽  
Vol 13 (2) ◽  
pp. 155-161 ◽  
Author(s):  
Haiqin Chen ◽  
Zhinan Xu ◽  
Naizheng Xu ◽  
Peilin Cen
Keyword(s):  
Coli Cell ◽  
High Level Expression ◽  
Free System ◽  
Cell Free System ◽  
E Coli ◽  
Rare Codons ◽  
High Level

scholarly journals Chaperone-Assisted Soluble Expression of a Humanized Anti-EGFR ScFv Antibody in E. Coli

2015 ◽  
Vol 5 (Suppl 1) ◽  
pp. 621-627 ◽  
Author(s):  
Kamal Veisi ◽  
Safar Farajnia ◽  
Nosratollah Zarghami ◽  
Hamid Reza Khoram Khorshid ◽  
Nasser Samadi ◽  
...  
Keyword(s):  
Soluble Expression ◽  
Scfv Antibody ◽  
E Coli

High level soluble expression and ATPase characterization of human heat shock protein GRP78

2017 ◽  
Vol 82 (2) ◽  
pp. 186-191
Author(s):  
Shuang Wu ◽  
Hongpeng Zhang ◽  
Miao Luo ◽  
Ke Chen ◽  
Wei Yang ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Soluble Expression ◽  
High Level ◽  
Human Heat Shock Protein

scholarly journals Molecular survey of mcr1 and mcr2 plasmid mediated colistin resistance genes in Escherichia coli isolates of animal origin in Iran

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Kayhan Ilbeigi ◽  
Mahdi Askari Badouei ◽  
Hossein Vaezi ◽  
Hassan Zaheri ◽  
Sina Aghasharif ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Companion Animals ◽  
Multidrug Resistant ◽  
Animal Origin ◽  
Colistin Resistance ◽  
E Coli ◽  
High Level ◽  
Farming Industry

Abstract Objectives The emergence of colistin-resistant Enterobacteriaceae from human and animal sources is one of the major public health concerns as colistin is the last-resort antibiotic for treating infections caused by multidrug-resistant Gram-negative bacteria. We aimed to determine the prevalence of the prototype widespread colistin resistance genes (mcr-1 and mcr-2) among commensal and pathogenic Escherichia coli strains isolated from food-producing and companion animals in Iran. Results A total of 607 E. coli isolates which were previously collected from different animal sources between 2008 and 2016 used to uncover the possible presence of plasmid-mediated colistin resistance genes (mcr-1 and mcr-2) by PCR. Overall, our results could not confirm the presence of any mcr-1 or mcr-2 positive E. coli among the studied isolates. It is concluded that despite the important role of food-producing animals in transferring the antibiotic resistance, they were not the main source for carriage of mcr-1 and mcr-2 in Iran until 2016. This study suggests that the other mcr variants (mcr-3 to mcr-9) might be responsible for conferring colistin resistance in animal isolates in Iran. The possible linkage between pig farming industry and high level of mcr carriage in some countries needs to be clarified in future prospective studies.

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