Uzgoj i permeabilizacija stanica Ralstonia eutropha H16

undergraduate thesis

• Title (english): Cultivation and permeabilization of Ralstonia eutropha H16 cells
• Author: Korina Krog
• Mentor: Ana Jurinjak Tušek (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Process Engineering); (Laboratory for Measurement, Regulation and Automatisation); Zagreb
• Defense date and country: 2024-07-16, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Kako bi se smanjila upotreba fosilnih goriva kao izvora energije. potiče se primjena zelenih alternativnih goriva, poput vodika. Posebno je zanimljiva proizvodnja biovodika koja se temelji na razgradnji organskih tvari (npr. organski otpad) primjenom mikroorganizama. U ovom radu analiziran je utjecaj sastava hranjive podloge (udio fruktoze i glicerola) na rast bakterijskog soja Ralstonia eutropha H16 koji metabolički proizvodi četiri hidrogenaze, obećavajuće biokatalizatore u proizvodnji biovodika. Također je analizirana učinkovitost šest različitih sredstava (EDTA, Triton X-100, SDS, CTAB, PEG i BSA) na permeabilizaciju stanica R. eutropha H16 kako bi se u što većoj koncentraciji oslobodili poželjni enzimi hidrogenaze. Rezultati pokazuju da je najveći prinos koncentracije biomase, od 0,470 ± 0,134 g/L, postignut u eksperimentu s 4 g/L otopine glicerola u hranjivoj podlozi. Nadalje, najveća koncentracija proteina, od 25,536 ± 1,102 mg/mL, oslobođena je koristeći CTAB kao sredstva za permeabilizaciju. Permeabilizat dobiven primjenom Triton X-100 pokazao je najveću enzimsku aktivnost zbog specifičnog načina interakcije s površinskim strukturama stanica.

Abstract (english)

In order to reduce the use of fossil fuels as an energy source, the use of green alternative fuels, such as hydrogen, is being promoted. Of particular interest is the production of biohydrogen, which is based on the decomposition of organic substances (e.g. organic waste) by microorganisms. In this work, the influence of the composition of the culture medium (proportion of fructose and glycerol) on the growth of the bacterial strain Ralstonia eutropha H16, which produces four hydrogenases, promising biocatalysts for the production of biohydrogen, was analyzed. The efficacy of six different agents (EDTA, Triton X-100, SDS, CTAB, PEG and BSA) on the permeabilization of R. eutropha H16 cells was also analyzed to release the desired hydrogenase enzymes at the highest possible concentration. The results show that the highest biomass concentration of 0.470 ± 0.134 g/L was achieved in the experiment with 4 g/L glycerol solution in the culture medium. In addition, the highest protein concentration of 25.536 ± 1.102 mg/ml was released when CTAB was used as a permeabilizing agent. The permeabilizate obtained with Triton X-100 showed the highest enzymatic activity due to the specific nature of the interaction with the surface structures of the cells.

• Keywords: biovodik; Ralstonia eutropha H16; permeabilizacija
• Keywords (english): biohydrogen; Ralstonia eutropha H16; permeabilization
• Language: croatian
• URN:NBN: urn:nbn:hr:159:400701
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-11-13 10:18:02