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Razvoj mikropretočne naprave z imobiliziranimi sporami Bacillus subtilis na osnovi magnetnega polja
ID Sotlar, Samo (Author), ID Žnidaršič Plazl, Polona (Mentor) More about this mentor... This link opens in a new window

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Abstract
Glavni izzivi biokatalitskih procesov so razvoj zelo aktivnih, robustnih in stabilnih biokatalizatorjev, učinkovita regeneracija in preprečevanje deaktivacije biokatalizatorja. Magnetni delci predstavljajo prilagodljiv nosilec za imobilizacijo encimov ali celic, saj imajo veliko specifično površino z možnostjo funkcionalizacije in omogočajo enostavno imobilizacijo biokatalizatorja z zunanjim magnetnim poljem. V pričujočem delu smo razvili mikropretočno napravo, v kateri smo učinkovito imobilizirali bakterijske spore s pomočjo magnetnih delcev. Preučili smo vpliv pH in različnih magnetnih delcev na lakazno aktivnost spor. Prav tako smo preučili vpliv pH in različnega razmerja med količino magnetnih delcev in spor na uspešnost imobilizacije spor v magnetnem polju. Pri optimalnih pogojih, ki so bili pH imobilizacije 2,5, magnetni delci MagP-NH2 in masno razmerje magnetnih delcev in spor 2,2, je bil izkoristek imobilizacije v pretočnem sistemu 97 %, zadržana lakazna aktivnost spor z magnetnimi delci MagP-NH2 v šaržnem sistemu pri pH 4 pa 88 %. Mikroreaktor z imobiliziranimi sporami v magnetnem polju smo uporabili za oksidacijo 2,2'-azino-bis(3-etilbenzotiazolin)-6-sulfonske kisline. Med potekom pretočnih reakcij smo ugotovili, da se v 5 urah procesa v reaktorju zadrži več kot 83 % spor. Koncentracijo produkta na izstopu iz mikroreaktorja smo spremljali izven sistema z UV/VIS spektrofotometrom, kot tudi z integriranim miniaturiziranim pretočnim spektrofotometrom, ki omogoča pridobivanje podatkov v realnem času.

Language:Slovenian
Keywords:Imobilizacija, spore Bacillus subtilis, magnetni delci, biotransformacija, mikropretočni sistemi
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2022
PID:20.500.12556/RUL-142024 This link opens in a new window
COBISS.SI-ID:134273539 This link opens in a new window
Publication date in RUL:17.10.2022
Views:707
Downloads:159
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Secondary language

Language:English
Title:Development of a magnetic field-bassisted microfluidic device with immobilized Bacillus subtilis spores
Abstract:
The main challenges of biocatalytic processes are the development of highly active, robust and stable biocatalysts, efficient regeneration and prevention of biocatalyst deactivation. Magnetic particles represent a flexible carrier for the immobilization of enzymes or cells, as they have a large specific surface area with the possibility of functionalization and enable easy immobilization of the biocatalyst with an external magnetic field. In the present work, we developed a microfluidic device in which we effectively immobilized bacterial spores with the help of magnetic particles. We examined the influence of pH and different magnetic particles on the laccase activity of spores. We also studied the effect of pH and different ratios between the amount of magnetic particles and spores on the immobilization efficiency in a magnetic field. At the optimal conditions, which were immobilization pH 2,5, MagP-NH2 magnetic particles and the mass ratio of magnetic particles to spores 2,2, the immobilization efficiency in the flow system was 97 %, the retained laccase activity of spores with MagP-NH2 magnetic particles was 88 % for the batch system at pH 4. A microreactor with immobilized spores in a magnetic field was used for the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid. During the flow reactions, we found that more than 83 % of the spores are retained in the reactor within 5 hours of the process. The concentration of the product at the outlet of the microreactor was monitored outside the system with a UV/VIS spectrophotometer, as well as with an integrated miniaturized flow spectrophotometer, which enables data acquisition in real time.

Keywords:Immobilization, Bacillus subtilis spores, magnetic particles, biotransformation, microfluidic systems

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