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Abstract

The aim of the study was to assess the feasibility of utilizing sodium alginate biopolymer as animmobilization carrier for laccase in the removal of indigo carmine (IC), an anionic dye. The main goal of this work was to optimize the decolourization process by selecting the appropriate immobilized enzyme dose per 1 mg of dye, as well as the process temperature. The effective immobilization of laccase using sodium alginate as a carrier was confirmed by Raman spectroscopy. An analysis of the size and geometric parameters of the alginate beads was also carried out. Tests of IC decolourization using alginate-laccase beads were conducted. Applying the most effective dose of the enzyme (320 mg of enzyme/1 mg of IC) made it possible to remove 92.5% of the dye over 40 days. The optimal temperature for the IC decolourization process, using laccase immobilized on sodium alginate, was established at 30-40ºC. The obtained results indicate that laccase from Trametes versicolor immobilized on sodium alginate was capable of decolourizing the tested dye primarily based on mechanism of biocatalysis.
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Bibliography

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Authors and Affiliations

Małgorzata Białowąs
1
ORCID: ORCID
Beata Kończak
1
Stanisław Chałupnik
1
Joanna Kalka
2
Magdalena Cempa
1
ORCID: ORCID

  1. Central Mining Institute – National Research Institute, Katowice, Poland
  2. Environmental Biotechnology Department, Faculty of Energy and Environmental Engineering,The Silesian University of Technology, Poland
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Abstract

The herbicide atrazine was incorporated in the granules manufactured in the process of tumbling agglomeration to obtain controlled release (CR) formulations. The formulations contained bentonite as a CRmatrix forming agent (960–974 g/kg of dry granules), atrazine (10 g/kg), citric acid (3.2 g/kg), and sodium alginate as a matrix binder and a release modifier (12.8–26.8 g/kg). The release characteristics of atrazine were studied by immersion of the granules in static water. The effects of formulations on atrazine transport through soil were studied using model soil columns irrigated with water. The release of atrazine from CR granules into water was affected by increasing the alginate concentration in a particular formulation because the time necessary for the release of 50% of the active ingredient was longer for the granules containing a higher amount of alginate. The CR formulations significantly reduced the amount of atrazine leached to the soil surface horizon in comparison with the commercial water suspension of the herbicide.

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Authors and Affiliations

Tomasz Małyszka
Tomasz Jankowski
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Abstract

The decolourization of Turquoise Blue HFG by immobilized cells of Lysinibacillus fusiformis B26 was investigated. Cells of L. fusiformis B26 were immobilized by entrapment in agar and calcium alginate matrices and attached in pumice particles. The effects of operational conditions (e.g., agar concentrations, cell concentrations, temperature, and inoculum amount) on microbial decolourization by immobilized cells were investigated. The results revealed that alginate was proven to be the best as exhibiting maximum decolourization (69.62%), followed by agar (55.55%) at 40°C. Pumice particles were the poorest. Optimum conditions for agar matrix were found: concentration was 3%, cell amount was 0.5 g and temperature was 40°C (55.55%). Ca-alginate beads were loaded with 0.5, 1.0 and 2.0 g of wet cell pellets and the highest colour removal activity was observed with 2.0 g of cell pellet at 40°C for alginate beads. Also, 0.5 and 1.0 g of pumice particles that were loaded with 0.25 and 0.5 g of cell pellets respectively were used and the results were found very similar to each other.

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Authors and Affiliations

Nazime Mercan Dogan
Tugba Sensoy
Gulumser Acar Doganli
Naime Nur Bozbeyoglu
Dicle Arar
Hatice Ardag Akdogan
Merve Canpolat
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Abstract

As part of the work, experiments were carried out on a laboratory scale to assess the effectiveness of the use of composite capsules based on halloysite and sodium alginate for the adsorption of copper from rainwater. The halloysite was subjected to acid activation prior to the encapsulation process. The characteristics of the capsules obtained were determined by means of SEM surface imaging, nitrogen adsorption by the BET method and pH PZC measurement by the suspension method. Adsorption was studied using various operational parameters such as adsorbent dose, contact time, pH and concentration of copper ions in the rainwater. A high percentage of copper ions removal was demonstrated, i.e. 72% for halloysite (H), and 83% for activated halloysite (HA) for a dose of 2.0 g/L. Adsorption of Cu (II) was consistent with pseudo-second order kinetics. The adsorbents showed a high adsorption capacity at the level of 11.03 mg/g, determined by the Langmuir isotherm model. This model fit well with the experimental data.
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Authors and Affiliations

Anna Marszałek
1
ORCID: ORCID

  1. Silesian University of Technology, Gliwice, Poland
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Abstract

Cell encapsulation seems to be a promising tool in tissue engineering. However, it has been shown to have several limitations in terms of long-term cell cultures due to an insufficient oxygen supply. In this study we propose the use of novel microcapsules designed for long-term cell culture consisting of an alginate shell and perfluorocarbon (PFC) core, which works as a synthetic oxygen carrier and reservoir. The influence of PFC presence in the culture as well as the size of structures on cell metabolism was evaluated during 21-day cultures in normoxia and hypoxia. We showed significant improvement in cell metabolism in groups where cells were encapsulated in hydrogel structures with a PFC core. The cells maintained a typical metabolism (oxidative phosphorylation) through all 21 days of the culture, overcoming the oxygen supply shortage even in large structures (diameter ¡ 1 mm). Applying PFC in alginate matrices can improve cell metabolism and adaptation in long-term cell cultures.
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Authors and Affiliations

Agata Stefanek
1
ORCID: ORCID
Aleksandra Kulikowska-Darłak
1
ORCID: ORCID
Karolina Bogaj
1
Aleksandra Nowak
1
Joanna Dembska
1
Tomasz Ciach
1
ORCID: ORCID

  1. Biomedical Engineering Laboratory, Faculty of Chemical and Process Engineering, Warsaw University of Technology, 00-645 Warsaw, Poland
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Abstract

Developing an effective and safe cancer therapy could significantly reduce the number of deaths and improve the quality of life of treated patients. Nowadays medicine has developed a wide range of anticancer chemotherapeutics but at the same time there is a lack of effective drug delivery methods. Therefore, the development of the targeted drug delivery system which will selectively release drug into the cancer cells is a key challenge of modern medicine.
The main aim of the presented research was to investigate the targeting effect of a drug delivery system based on the controlled release of dextran nanoparticles containing the anticancer drug – doxorubicin from the alginate microspheres coated with chitosan multilayers.
During the research the physicochemical properties of the alginate microspheres and its stability in the physiological environment were investigated. Moreover, the kinetics of the nanoparticles with doxorubicin release from the alginate microspheres covered with chitosan multilayers was characterized, depending on the thickness of the chitosan layer. Further, the cytotoxicity study of the alginate microspheres covered with chitosan multilayer and containing nanoparticles was performed to determine the therapeutic effect of the released nanoparticles with doxorubicin on the HeLa cells during the in vitro cell culture.
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Authors and Affiliations

Aleksandra Kulikowska-Darłak
1
ORCID: ORCID
Agata Stefanek
1
ORCID: ORCID
Iga Wasiak-Wojasińska
1
ORCID: ORCID
Paulina Wiechecka-Ożdżyńska
1
ORCID: ORCID
Tomasz Ciach
1
ORCID: ORCID

  1. Faculty of Chemical and Process Engineering, Warsaw University of Technology, Warynskiego 1 Street, 00-645 Warsaw, Poland
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Abstract

The process of obtaining alginate microspheres (AMs) by emulsification method was optimized by applying statistical analysis software. Ten batches of microspheres were prepared using the fractional plan 3 (K-p). AMs were obtained with two different methods: an ultrasonic homogenization (UH) process and a rotor-stator mechanical homogenization (MH). The amount of a cross-linking agent (CaCl 2), calcium chloride rate addition, and the sonication amplitude (UH) or the speed of rotor rotation (MH) were selected as formulation variables. All the batches were evaluated in terms of stability and size of the alginate microspheres. Approximation profiles were developed. As a result of the conducted research, stable alginate microspheres with sizes ranging from 10 to 30 micrometres were obtained. The obtained results showed that the quality of AMs was mainly affected by the concentration and the rate of calcium chloride addition into the system. Therefore, the role of calcium ions in the mechanisms of shell structuring was discussed. Lactobacillus casei bacteria were encapsulated into the batches found to be optimum. The high encapsulation efficiency (EE) of the bacteria (72-94%) depending on the form) and their viability over time were obtained. The model developed in the study can be effectively utilized to achieve the AMs formulations.
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Authors and Affiliations

Anna Łętocha
1
Alicja Michalczyk
2
ORCID: ORCID
Małgorzata Miastkowska
1
ORCID: ORCID
Elżbieta Sikora
1
ORCID: ORCID

  1. Cracow University of Technology, Department of Chemical Engineering and Technology,Cracow, Poland
  2. Lukasiewicz - Research Network-Institute of Industrial Organic Chemistry, Warsaw, Poland
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Abstract

This study aims at the immobilization and characterization of thermoalkalophilic lipases produced recombinantly from Bacillus thermocatenulatus BTL2 and Bacillus pumilus MBB03. For this purpose, immobilization of the produced enzymes in calcium-alginate@gelatin (Ca–Alg@gelatin) hydrogel beads, immobilization optimization and characterization measurements of the immobilized-enzyme hydrogels were conducted. Optimum temperature and pH values were determined for B. thermocatenulatus and B. pumilus MBB03 immobilized-enzyme hydrogels (60–70 °C, 55 °C and pH 9.5, pH 8.5). Thermal stability was determined between 65 °C and 60 °C of B. thermocatenulatus and B. pumilus MBB03 immobilized enzymes, respectively. The pH stability was determined between pH 7.0–11.0 at +4°C and pH 8.0–10.0 at +4 °C, respectively.
In conclusion, the entrapment technique provided controlled production of small diameter hydrogel beads (~ 0:19 and ~ 0:29) with negligible loss of enzyme. These beads retained high lipase activity at high pH and temperature. The activity of Ca–Alg@gelatin-immobilized lipase remained relatively stable for up to three cycles and then markedly decreased. With this enzyme immobilization, it may have a potential for use in esterification and transesterification reactions carried out in organic solvent environments. We can conclude that it is one of the most promising techniques for highly efficient and economically competent biotechnological processes in the field of biotransformation, diagnostics, pharmaceutical, food and detergent industries.
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Authors and Affiliations

Kezban Yildiz Dalginli
1
ORCID: ORCID
Onur Atakisi
2
ORCID: ORCID

  1. Department of Chemistry and Chemical Processing Technologies, Kars Vocational High School Kafkas University, Kars, Turkey
  2. Department of Chemistry, Faculty Science and Letter, Kafkas University, Kars, Turkey
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Abstract

Several bacteria that are associated with macroalgae can use phycocolloids as a carbon source. Strain INACH002, isolated from decomposing Porphyra (Rhodophyta), in King George Island, Antarctica, was screened and characterized for the ability to produce agarase and alginate-lyase enzymatic activities. Our strain INACH002 was identified as a member of the genus Flavobacterium, closely related to Flavobacterium faecale, using 16S rRNA gene analysis. The INACH002 strain was characterized as psychrotrophic due to its optimal temperature (17°C) and maximum temperature (20°C) of growth. Agarase and alginate-lyase displayed enzymatic activities within a range of 10°C to 50°C, with differences in the optimal temperature to hydrolyze agar (50°C), agarose (50°C) and alginate (30°C) during the first 30 min of activity. Strain Flavobacterium INACH002 is a promising Antarctic biotechnological resource; however, further research is required to illustrate the structural and functional bases of the enzymatic performance observed during the degradation of different substrates at different temperatures.
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Authors and Affiliations

Paris Lavín
Cristian Atala
Jorge Gallardo-Cerda
Marcelo Gonzalez-Aravena
Rodrigo De La Iglesia
Rómulo Oses
Cristian Torres-Díaz
Nicole Trefault
Marco A. Molina-Montenegro
H. Dail Laughinghouse IV
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Abstract

Shoot tips excised from shoot culture of Salvia officinalis were encapsulated in 2% or 3% (w/v) sodium alginate and exposed to 50 mM calcium chloride for complexation. Immediately or after 6, 12 or 24 weeks of storage at 4°C, the synthetic seeds were cultured for 6 weeks on half-strength MS medium supplemented with indole-3-acetic acid (IAA) (0.1 mg/l) and solidified with 0.7% agar. The frequency of shoot and root emergence from encapsulated shoot tips was affected by the concentrations of sodium alginate and additives in the gel matrix (sucrose, gibberellic acid, MS nutrient medium) as well as duration of storage. The frequency of shoot and root induction of non-stored synthetic seeds was highest with shoot tips encapsulated with 2% sodium alginate containing 1.5% sucrose and 0.5 mg/l gibberellic acid (GA3). Shoot tips maintained their viability and ability to develop shoots even after 24 weeks of storage when they were encapsulated in 3% alginate with 1/3 MS medium, sucrose (1.5%) and GA3 (0.25 mg/l). Root formation tended to decrease with storage time. Overall, 90% of the plantlets derived from stored and non-stored synthetic seeds survived in the greenhouse and grew to phenotypically normal plants. This procedure can enable the use of synthetic seed technology for germplasm conservation of S. officinalis, a plant species of high medical and commercial value.

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Authors and Affiliations

Izabela Grzegorczyk
Halina Wysokińska

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