scholarly journals Extractive purification of recombinant thermostable lipase from fermentation broth of Escherichia coli using an aqueous polyethylene glycol impregnated resin system

3 Biotech ◽  
2018 ◽  
Vol 8 (6) ◽  
Author(s):  
Joo Shun Tan ◽  
Sahar Abbasiliasi ◽  
Arbakariya B. Ariff ◽  
Hui Suan Ng ◽  
Mohamad Hafizi Abu Bakar ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Fermentation Broth ◽  
Resin System ◽  
Thermostable Lipase ◽  
Impregnated Resin

Partitioning behavior of recombinant lipase in Escherichia coli by ionic liquid-based aqueous two-phase systems

RSC Advances ◽  
2016 ◽  
Vol 6 (86) ◽  
pp. 82571-82580 ◽  
Author(s):  
Muhammad Hakimi Hadzir ◽  
Sahar Abbasiliasi ◽  
Arbakariya B. Ariff ◽  
Siti Baidurah Yusoff ◽  
Hui Suan Ng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ionic Liquid ◽  
Ionic Liquids ◽  
Fermentation Broth ◽  
Two Phase ◽  
E Coli ◽  
Recombinant Lipase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems ◽  
Partitioning Behavior

Evaluations of ILATPSs were performed with a variety of ionic liquids and salts as phase components to figure out their competencies in the recovery of lipase from a fermentation broth of E. coli using banana waste as a substrate.

Recombinant strains producing thermostable lipase from Thermomyces lanuginosus and their use in heterogeneous biocatalysis, including in the processes of low-temperature synthesis of esters

2021 ◽  
Vol 37 (5) ◽  
pp. 5-19
Author(s):  
M.B. Pykhtina ◽  
L.V. Perminova ◽  
G.A. Kovalenko
Keyword(s):  
Escherichia Coli ◽  
Fatty Acids ◽  
Pichia Pastoris ◽  
Substrate Specificity ◽  
Carbon Aerogel ◽  
Operational Stability ◽  
Thermomyces Lanuginosus ◽  
Thermostable Lipase ◽  
Recombinant Strains ◽  
Heterogeneous Biocatalysts

Abstract-This work was devoted to the construction of recombinant strains Escherichia coli BL21 (DE3) and Pichia pastoris X33, producing a 1,3-specific thermostable lipase from Thermomyces lanuginosus. The sequences of two lipase genes were optimized for expression in bacteria and methylotrophic yeasts, then synthesized and cloned into the corresponding expression vectors. As a result of genetic engineering manipulations, E. coli and P. pastoris strains were constructed that efficiently produced recombinant lipase from T. lanuginosus, which accumulated in the cytoplasm in an amount of 30-40% of the total cellular protein. Recombinant P. pastoris clones secreted lipase into the nutrient medium at a concentration of at least 1 g/L. Lipases produced by the recombinant clones, designated as rE.coli/lip and rPichia/lip, respectively, contained a six-histidine sequence (-His6) in the C-terminal region. The resulting lipases were immobilized on/in solid inorganic supports in order to develop heterogeneous biocatalysts (HB) for the enzymatic conversion of triglycerides and fatty acids. The rPichia/lip enzyme was adsorbed on mesoporous silica and macroporous carbon aerogel. The properties of the prepared HB, their enzymatic activity, substrate specificity and operational stability were studied in the reaction of esterification of fatty acids with aliphatic alcohols in organic solvents at 20 ± 2°C. It was found that immobilized lipases had a relatively wide substrate specificity, as well as high operational stability, and the prepared HB almost completely retained their high esterifying activity for several tens of reaction cycles. Key words: Escherichia coli, Pichia pastoris, recombinant strains-producers, Thermomyces lanuginosus lipase gene, immobilization, biocatalysts, esterification The authors are grateful to V. L. Kuznetsov for the provided samples of carbon aerogel and A. V. Ryabchenko for gene-engineering manipulation aimed at obtaining the recombinant rE. coli strain, a producer of the rE.coli/lip enzyme. The work was carried out under the Project on Fundamental Research within the framework of a state assignment to the Institute for Catalysis "Catalysts and Processes of Renewable Raw Material Conversion" (no. 0239-2021-0005).

scholarly journals Crystallization and preliminary X-ray crystallographic analysis of human myotubularin-related protein 1

2015 ◽  
Vol 71 (3) ◽  
pp. 261-265 ◽  
Author(s):  
Seoung Min Bong ◽  
Seung Won Yang ◽  
Ji-Woong Choi ◽  
Seung Jun Kim ◽  
Byung Il Lee
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Synchrotron Radiation ◽  
Crystallographic Analysis ◽  
Asymmetric Unit ◽  
Related Protein ◽  
Unit Cell Parameters ◽  
Solvent Content ◽  
X Ray ◽  
Cell Parameters

Myotubularin-related protein 1 is a phosphatase that dephosphorylates phospholipids such as phosphatidylinositol 3-phosphate or phosphatidylinositol 3,5-bisphosphate. In this study, human MTMR1 was overexpressed inEscherichia coli, purified and crystallized at 277 K using polyethylene glycol 20 000 as a precipitant. Diffraction data were collected to 2.0 Å resolution using synchrotron radiation. The crystals belonged to space groupP1, with unit-cell parametersa= 67.219,b= 96.587,c= 97.581 Å, α = 87.597, β = 86.072, γ = 77.327°. Assuming the presence of four molecules in the asymmetric unit, the calculated Matthews coefficient value was 2.61 Å3 Da−1and the corresponding solvent content was 52.9%.

scholarly journals Effect of polyethylene glycol in plasmid DNA solution on transformation of CaCl2-treated Escherichia coli cells.

1984 ◽  
Vol 48 (3) ◽  
pp. 657-662 ◽  
Author(s):  
Michio HIMENO ◽  
Toshihiro SHIBATA ◽  
Yoshio KAWAHARA ◽  
Yuhji HANAOKA ◽  
Tohru KOMANO
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Plasmid Dna ◽  
Escherichia Coli Cells

scholarly journals EFEK ANTIBAKTERI SENYAWA METABOLIT SEKUNDER YANG DIPRODUKSI OLEH KAPANG ENDOFITIK AT 32 DARI Artemisia annua

2017 ◽  
Vol 1 (1) ◽  
pp. 61
Author(s):  
Djadjat Tisnadjaja
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Secondary Metabolite ◽  
Active Compound ◽  
Artemisia Annua ◽  
Fermentation Broth ◽  
Research Result ◽  
Stirred Tank ◽  
Zone Formation ◽  
Major Health Problem

Antibacterial Effect of Secondary Metabolite Compound Produced by Endophyte Mould AT 32Infection diseases such as malaria, dengue hemorrhagic fever, and tuberculosis still counted as one of major health problem in Indonesia. On the other hand Indonesia has mega biodiversity including plant and microbes potentially used for the production of antibacterial or lead compound for antibiotic. One of a potential resources is Artemisia annua. This plant is already known as a secondary metabolite compound artemisinin producer, an active compound against malaria virus. This research work  was aimed to study the possible used of endophyte microbe isolated from Artemisia annua to produce antibacterial active compound through a fermentation process. Research result shown that extract of fermentation broth, either fermentation was conducted in Erlenmeyer flask or two liters stirred tank reactor, gave antimicrobial activity. It is shown by the clearing zone formation on the dish inoculated with bacterial seed.  Two bacterial strains Staphylococcus aureus ATCC 25923 and Escherichia coli BCC 2210 were used for this study. The biggest clearing zone diameter formed was 20 mm for inhibition of Staphylococcus aureus and 9.5 mm for Eschericia coli, those were obtained by using extract of fermentation broth of two liters stirred tank reactor.Keywords  :  Artemisis annua, antimicrobial, endophyte, fermentation, secondary metabolite ABSTRAK         Penyakit infeksi seperti malaria, demam berdarah dan tuberculosis (TBC) masih merupakan masalah kesehatan utama di Indonesia. Pada sisi lain Indonesia memiliki mega biodiversitas termasuk tanaman dan mikroba yang berpotensi untuk dimanfaatkan dalam produksi antibakteri atau  senyawa aktif untuk antibiotik. Penelitian ini dilakukan untuk mempelajari kemungkinan pemanfaatan mikroba endofitik yang diisolasi dari tumbuhan Artemisia annua untuk memproduksi senyawa aktif antibakteri melalui proses fermentasi. Hasil penelitian menunjukkan bahwa ekstrak dari cairan fermentasi, baik ketika fermentasi dilakukan dalam skala Erlenmeyer maupun fermentor stirred tank skala 2 liter, menunjukkan adanya aktivitas antimikroba. Aktivitas antimikroba ini ditunjukkan dengan terbentuknya zona jernih pada petri dish yang diinokulasi dengan bakteri. Dalam hal ini digunakan dua galur bakteri patogen yaitu Staphylococcus aureus ATCC 25923 dan Escherichia coli BCC 2210. Diameter zona jernih terbesar yang terbentuk yaitu 20 mm untuk inhibisi dari Staphylococus aureus dan 9,5 mm untuk penghambatan Escherichia coli keduanya diperoleh dengan pemberian ekstrak cairan fermentasi yang dilakukan dengan fermentor stirred tank skala 2 liter.Kata kunci : Artemisis annua, anti mikroba, endophyte, fermentasi, metabolit sekunder

scholarly journals Effect of different aeration and polyethylene glycol concentration on growth of Escherichia coli DH5α in a 1L bioreactor

2018 ◽  
Author(s):  
Wenfa Ng
Keyword(s):  
Escherichia Coli ◽  
Polyethylene Glycol ◽  
Cell Growth ◽  
Growth Medium ◽  
Growth Rates ◽  
Shake Flask ◽  
Foam Formation ◽  
Biomass Formation ◽  
E Coli ◽  
Toxicity Effect

Large culture volume in bioreactor necessitates aeration for providing sufficient oxygen for cell growth. Thus, extend of aeration and amount of anti-foam needed for suppressing foam formation are key parameters determining the success of bioreactor fermentation. Specifically, while aeration provides more oxygen for powering cellular metabolism and could lead to faster growth rate and more efficient metabolism, it also introduces greater shear stress, mixing and foam formation. On the other hand, anti-foaming agents such as polyethylene glycol (PEG) could exert a toxicity effect on cells as well as introducing increased osmolarity and viscosity that could hamper cell growth. In this preliminary study, effect of different PEG concentrations and extent of aeration on growth of Escherichia coli DH5α (ATCC 53868) in a 1L bioreactor at 37 oC with LB Lennox growth medium was investigated. Experiment results revealed that E. coli DH5α growth in bioreactor at 1 VVM aeration with 1 g/L PEG was faster than that in a 250 mL glass shake flask, and with greater secretion of alkaline metabolites. Similar optical density obtained between bioreactor and shake flask cultivation pointed to the maximized utilization of growth medium nutrients for biomass formation. Increase in bioreactor aeration to 3 VVM at 1 g/L PEG, however, resulted in increased secretion of acidic metabolites into the culture broth while allowing similar maximal optical density to be obtained compared to aeration of 1 VVM at 1 g/L PEG. This indicated that E. coli DH5α was able to adapt to physiological impacts from increased aeration and highlighted that no significant metabolic energy was diverted from biomass formation. Finally, increase in PEG concentration to 10 g/L from 1 g/L did not introduce additional toxicity effect given that growth profile of E. coli DH5α under the two PEG concentrations overlapped each other. However, observations of reduced secretion of acidic metabolites at the outset of growth in 10 g/L PEG pointed to physiological impacts that did not affect growth rates and biomass formation. Collectively, E. coli DH5α was able to tolerate enhanced aeration of 3 VVM and 10 g/L PEG anti-foam without significant detrimental impacts on growth rates and biomass formation.

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