20.500.12556/RUL-149088 Prednosti in slabosti uporabe bakterije Bacillus subtilis kot celične tovarne za produkcijo heterolognih proteinov v industriji Advantages and disadvantages of using Bacillus subtilis as cell factory for the production of heterologous proteins in industry Bacillus subtilis je Gram pozitivna bakterija z velikim potencialom za celično tovarno heterolognih proteinov. Že od prejšnjega stoletja se jo uporablja za proizvodnjo encimov in drugih učinkovin, ima dobro poznan genom in razvita orodja za genetske modifikacije genoma. B. subtilis je zelo dovzetna za sprejem tuje DNK, poleg kromosomske in plazmidne DNK sprejme tudi krajše PCR fragmente. Za spreminjanje genoma uporabljajo tehnike CSM, SSR in CRISPR-Cas9. Ima izjemno prilagodljiv metabolizem, zato lahko raste ne enostavnih in poceni substratih. Izloča velike količine produkta, največ preko sekrecijskih poteh Sec in Tat direktno v medij, zato je postopek čiščenja lažji in cenejši. Prednost uporabe B. subtilis je tudi status GRAS, ne izloča endotoksinov ter eksotoksinov in ne proizvaja inkluzijskih telesc. Poleg prednosti ima tudi nekaj slabosti. Ni sposobna tvoriti disulfidnih vezi, saj so encimi nespecifični in počasni. Največjo težavo predstavljajo encimi proteaze, ki razgradijo nastale heterologne proteine in zmanjšujejo donos proizvodnje. Najbolj znani produkti B. subtilis so humani rastni faktor in industrijski encimi amilaza, laktaza, celulolitični encimi in ksilanaza. Bacillus subtilis is a Gram-positive bacterium with great potential as a cellular factory for heterologous proteins. It has been used since the last century to produce enzymes and other active substances, it has a well-known genome and well developed tools for genetic genome modification. B. subtilis is very susceptible to accepting foreign DNA, accepting short PCR fragments in addition to chromosomal and plasmid DNA. It has an extremely flexible metabolism and can therefore grow on simple and inexpensive substrates. It excretes large amounts of product, mostly via the Sec and Tat secretion pathways, directly into the medium, making the purification process easier and cheaper. B. subtilis also has the advantage of GRAS status, it does not secrete endotoxins and exotoxins and does not produce inclusion bodies. In addition to the advantages, it also has some disadvantages. It is not able to form disulfide bonds as the enzymes are non-specific and slow. The biggest problem is the protease enzymes, which degrade the resulting heterologous proteins and reduce the yield of production. The best known products of B. subtilis are human growth factor and the industrial enzymes amylase, laccase, cellulolytic enzymes and xylanase. Bacillus subtilis celična tovarna heterologni proteini sekrecijske poti manipulacija genoma Bacillus subtilis cell factory heterologous proteins secretion pathways genome manipulation true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2023-09-03 07:16:27 2023-09-03 07:16:29 2023-10-04 11:43:51 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 228232 163218691 34647.pdf 34647.pdf 1 B2AF547432EBC7EA113C124F18B53801 07a09d0b80337fbaf5c69d01ffd94b48a661907a9ffabf0d44c5c33833cfcab6 081b3732-4a19-11ee-af86-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=173475 Biotehniška fakulteta 0 0 0