scholarly journals Chemical and thermal stabilization of CotA laccase via a novel one-step expression and immobilization in muNS-Mi nanospheres

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tomás Pose-Boirazian ◽  
Gemma Eibes ◽  
Natalia Barreiro-Piñeiro ◽  
Cristina Díaz-Jullien ◽  
Juan M. Lema ◽  
...  

AbstractA methodology that programs eukaryotic or bacterial cells to encapsulate proteins of any kind inside micro/nanospheres formed by muNS-Mi viral protein was developed in our laboratory. In the present study such “in cellulo” encapsulation technology is utilized for immobilizing a protein with an enzymatic activity of industrial interest, CotA laccase. The encapsulation facilitates its purification, resulting in a cost-effective, one-step way of producing immobilized enzymes for industrial use. In addition to the ability to be recycled without activity loss, the encapsulated protein showed an increased pH working range and high resistance to chemical inactivation. Also, its activity was almost unaffected after 30 min incubation at 90 °C and 15 min at the almost-boiling temperature of 95 °C. Furthermore, the encapsulated laccase was able to efficiently decolorate the recalcitrant dye RB19 at room temperature.

2012 ◽  
Vol 78 (15) ◽  
pp. 5440-5443 ◽  
Author(s):  
Jae-Yeon Jeong ◽  
Hyung-Soon Yim ◽  
Ji-Young Ryu ◽  
Hyun Sook Lee ◽  
Jung-Hyun Lee ◽  
...  

ABSTRACTWe developed one-step sequence- and ligation-independent cloning (SLIC) as a simple, cost-effective, time-saving, and versatile cloning method. Highly efficient and directional cloning can be achieved by direct bacterial transformation 2.5 min after mixing any linearized vector, an insert(s) prepared by PCR, and T4 DNA polymerase in a tube at room temperature.


2010 ◽  
Vol 159 ◽  
pp. 303-306
Author(s):  
Sharmin Haq ◽  
Tasnuva Ashrafee ◽  
Mahmuda Begum ◽  
Tasmia Sharmin ◽  
Nirban Bhowmick ◽  
...  

The new emergent properties of nanoparticles include quantam size effect, nonlinear optical properties and thus the nanomaterials have extensive applications in microelectronic, bacteriostatic and catalytic or magnetic recording materials. Nanoparticles find potential applications in DNA detection and photo detection. Gold nano particles (AuNPs) are useful in nanoscale devices and systems due to its resistance to oxidation. In this paper a cost effective, versatile and simple one step room temperature synthesis of highly stable and freestanding Au quantum dots capped by TTAB (Tetradecyltrimethyl Amonium Bromide) in the range of 10-90 nm is reported. Ultraviolet- visible spectroscopy (UV-VIS), Fourier transformation infrared spectroscopy (FT-IR) was carried out to characterize these samples. Scanning Electron Microscopy (SEM) confirms the AuNPs sizes of about 80 and 88 nm.


2016 ◽  
Vol 9 (1) ◽  
pp. 80-95
Author(s):  
Agus Sudibyo ◽  
Sardjono Sardjono

Crude palm oil (CPO)is the richest natural plant source of carotenoids in terms of retinol (pro-vitamin A) equivalent, whereas palm oil mill effluent (POME) is generated from palm oil industry that contains oil and carotenes that used to be treated before discharge. Carotenoids are importance in animals and humans for the purpose of the enhancement of immune response, conversion of vitamin A and scavenging of oxygen radicals. This component has different nutritional  functions and benefits to humaan health. The growing interest in the other natural sources of beta-carotene and growing awareness to prevent pollution has stimulated the industrial use of CPO and POME as a raw material for carotenoids extraction. Various technologies of extraction and separation have been developed in order to recover of carotenoids.This article reports on various technologies that have been developed in order to recover of carotenoids from being destroyed in commercial refining of palm oil and effects of some various treatments on the extraction end separation for carotenoid from palm oil and carotenoids concentration. Principally, there are different technologies, and there is one some future which is the use of solvent. Solvent plays important role  in the most technologiest, however the problem of solvents which are used is that they posses potentiaal fire health and environmental hazards. Hence selection of the  most safe, environmentally friendly and cost effective solvent is important to design of alternative extraction methods.Chemical molecular product design is one of the methods that are becoming more popular nowadays for finding solvent with the desired properties prior to experimental testing.ABSTRAKMinyak sawit kasar merupakan sumber karotenoid terkaya yang berasal dari tanaman sawit sebagai senyawa yang sama dengan retinol atau pro-vitamin A; sedangkan limbah pengolahan minyak sawit dihasilkan dari industri pengolahan minyak sawit yang berisi minyak dan karotene yang perlu diberi perlakuan terlebih dahulu sebelum dibuang. Karotenoid merupakan bahan penting yang diperlukan pada hewan dan manusia guna memperkuat tanggapan terhadap kekebalan, konversi ke vitamin A dan penangkapan gugus oksigen radikal. Dengan berkembangnya ketertarikan dalam mencari beta-karotene yang bersumber dari alam lain dan meningkatnya kesadaran untuk mencegah adanya pencemaran lingkungan, maka mendorong suatu industri untuk menggunakan CPO dan POME sebagai bahan baku untuk diekstrak karotenoidnya. Berbagai macam teknologi guna mengekstrak dan memisahkan karotenoid telah dikembangkan untuk mendapatkan kembali karotenoidnya. Makalah ini melaporkan dan membahas berbagai jenis teknologi yang telah dikembangkan guna mendapatkan kembali senyawa karotenoid dari kerusakan di dalam proses pemurnian minyak sawit secara komersial dan pengaruh beberapa perlakuan terhadap ekstrasi dan pemisahan karotenoid dari minyak sawit dan konsentrasi karotenoidnya. Pada prinsipnya, berbagai teknologi yang digunakan untuk mengekstrak dan memisahkan karotenoid terdapat perbedaan, dan terdapat salah satu teknologi yang digunakan untuk esktrasi dan pemisahan karotenoid adalah menggunakan bahan pelarut. Pelarut yang digunakan mempunyai peranan yang penting dalam teknologi ekstrasi; namun pelarut yang digunakan untuk mengekstrak tersebut mempunyai persoalan karena berpotensi mengganggu kesehatan dan membahayakan cemaran lingkungan. Oleh karena itu, pemilihan jenis teknologi yang aman, ramah terhadap lingkungan dan biaya yang efektif untuk penggunaan pelarut merupakan hal penting sebelum dilakukan desain metode/teknologi alternatif untuk esktrasi karotenoid. Pola produk molekuler kimia merupakan salah satu metode yang saat ini menjadi lebih populer untuk mencari pelarut dengan sifat-sifat yang dikehendaki sebelum diujicobakan. Kata kunci :    karotenoid, ekstrasi, pemisahan, teknologi, minyak sawit kasar, limbah industri pengolahan sawit.


2020 ◽  
Vol 24 (4) ◽  
pp. 465-471 ◽  
Author(s):  
Zita Rádai ◽  
Réka Szabó ◽  
Áron Szigetvári ◽  
Nóra Zsuzsa Kiss ◽  
Zoltán Mucsi ◽  
...  

The phospha-Brook rearrangement of dialkyl 1-aryl-1-hydroxymethylphosphonates (HPs) to the corresponding benzyl phosphates (BPs) has been elaborated under solid-liquid phase transfer catalytic conditions. The best procedure involved the use of triethylbenzylammonium chloride as the catalyst and Cs2CO3 as the base in acetonitrile as the solvent at room temperature. The substrate dependence of the rearrangement has been studied, and the mechanism of the transformation under discussion was explored by quantum chemical calculations. The key intermediate is an oxaphosphirane. The one-pot version starting with the Pudovik reaction has also been developed. The conditions of this tandem transformation were the same, as those for the one-step HP→BP conversion.


2020 ◽  
Vol 16 (6) ◽  
pp. 795-799
Author(s):  
YongJin Li

Background: A simple, fast and economic analytical method for the determination of ethanol is important for clinical, biological, forensic and physico-legal purposes. Methods: Ni2+-NTA resin was used as an immobilization matrix for the simple one-step purification/ immobilization of his6-tagged ADH. Different alcohols with a concentration range of 0.5-50% V/V, namely methanol, ethanol and propanol were measured using prepared ADH enzyme thermistor. The ethanol content of Tsingtao beer was tested as a real sample containing alcohol. Reproducibility and stability of prepared ADH enzyme thermistor were also investigated by repeated measurements. Results: In comparison to the controlled pore glass (a common used support for the immobilization of enzyme) used in thermal biosensor, the use of Ni2+-NTA resin not only led to simple one-step purification/ immobilization by his6-tagged ADH binding to Ni2+-NTA resin, but also made the immobilizing supports reusable. The prepared biosensor can be used to determine ethanol and methanol by the calorimetric measurement. A linear range of 1 -32% (V/V) and 2-20% (V/V) was observed for ethanol and methanol, respectively. The detection limits were 0.3% (V/V) and 1% (V/V) for ethanol and methanol, respectively. The tested ethanol concentration of Tsingtao beer was 4.5% V/V, which is comparable with the labeled alcohol by volume (ABV) 4.80%. Conclusion: Ni2+-NTA resin, as an immobilization matrix in ET sensor, provides a simple one-step purification/immobilization for His6-tagged recombinase and a reusable immobilization matrix. The prepared biosensor exhibits good repeatability and stability. Such a new biosensor shows great promise for rapid, simple, and cost-effective analysis of ethanol and methanol, both in qualitative and in quantitative tests.


Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 893
Author(s):  
Olufunto T. Fanoro ◽  
Sundararajan Parani ◽  
Rodney Maluleke ◽  
Thabang C. Lebepe ◽  
Jose R. Varghese ◽  
...  

We herein report a facile, green, cost-effective, plant-mediated synthesis of gold nanoparticles (AuNPs) for the first time using Combretum erythrophyllum (CE) plant leaves. The synthesis was conducted at room temperature using CE leaf extract serving as a reducing and capping agent. The as-synthesized AuNPs were found to be crystalline, well dispersed, and spherical in shape with an average diameter of 13.20 nm and an excellent stability of over 60 days. The AuNPs showed broad-spectrum antibacterial activities against both pathogenic Gram-positive (Staphylococcus epidermidis (ATCC14990), Staphylococcus aureus (ATCC 25923), Mycobacterium smegmatis (MC 215)) and Gram-negative bacteria (Proteus mirabilis (ATCC 7002), Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATCC 13822), Klebsiella oxytoca (ATCC 8724)), with a minimum inhibition concentration of 62.5 µg/mL. In addition, the as-synthesized AuNPs were highly stable with exceptional cell viability towards normal cells (BHK- 21) and cancerous cancer cell lines (cervical and lung cancer).


2021 ◽  
Vol 12 (5) ◽  
pp. 645-649
Author(s):  
Yu Gu ◽  
Yuqing Qiao ◽  
Yusen Meng ◽  
Ming Yu ◽  
Bowu Zhang ◽  
...  

Herein, we report for the first time the synthesis of polypyrrole copolymers with good solvent-dispersibility under gamma-ray irradiation at room temperature in air.


Author(s):  
Francisco G. Cirujano ◽  
Nuria Martin ◽  
Neyvis Almora-Barrios ◽  
Carlos Martí-Gastaldo

Room temperature one-step synthesis of the peptide-based porous material with a periodic distribution of pockets decorated with lysine side chain active sites behaves as a heterogeneous organocatalyst. The pockets are...


Pharmaceutics ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 13
Author(s):  
Benzion Amoyav ◽  
Yoel Goldstein ◽  
Eliana Steinberg ◽  
Ofra Benny

Microfluidics research for various applications, including drug delivery, cell-based assays and biomedical research has grown exponentially. Despite this technology’s enormous potential, drawbacks include the need for multistep fabrication, typically with lithography. We present a one-step fabrication process of a microfluidic chip for drug dissolution assays based on a 3D printing technology. Doxorubicin porous and non-porous microspheres, with a mean diameter of 250µm, were fabricated using a conventional “batch” or microfluidic method, based on an optimized solid-in-oil-in-water protocol. Microspheres fabricated with microfluidics system exhibited higher encapsulation efficiency and drug content as compared with batch formulations. We determined drug release profiles of microspheres in varying pH conditions using two distinct dissolution devices that differed in their mechanical barrier structures. The release profile of the “V” shape barrier was similar to that of the dialysis sac test and differed from the “basket” barrier design. Importantly, a cytotoxicity test confirmed biocompatibility of the printed resin. Finally, the chip exhibited high durability and stability, enabling multiple recycling sessions. We show how the combination of microfluidics and 3D printing can reduce costs and time, providing an efficient platform for particle production while offering a feasible cost-effective alternative to clean-room facility polydimethylsiloxane-based chip microfabrication.


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