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Development of cellulose-degrading lactic acid bacterium Lactococcus cremoris by genetic engineering
ID Štravs, Petra (Avtor), ID David, Hélène (Avtor), ID Fierobe, Henri-Pierre (Avtor), ID Perret, Stéphanie (Avtor), ID Berlec, Aleš (Avtor)

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Izvleček
Cellulose is one of the most abundant potential sources of carbon for sustainable microbial production of biochemicals. Lactic acid bacteria that produce a range of valuable metabolites are generally unable to grow on cellulose. Here, we aimed to develop the lactic acid bacteria Lactococcus cremoris for growth on cellulosic substrate. Genes encoding cellulases Cel5I, Cel9A and Cel5H from different cellulolytic bacteria were introduced into L. cremoris NZ9000 strain. The genes were designed for constitutive expression, with the produced cellulases being either secreted or displayed on the cell surface. Four promoters for cellulase gene expression and two noncovalent anchors for surface display of cellulases were evaluated using immunoblotting, confocal microscopy and flow cytometry. With the most effective promoter PepN, the cells secreted milligram amounts of cellulases per liter culture. The highest cellulolytic activity on amorphous (phosphoric-acid swollen cellulose, PASC) and microcrystalline (Avicel) cellulose was observed for the secreted Cel5H and Cel5I cellulases, respectively. The major cellodextrin produced by Cel5H and Cel5I was cellobiose, as determined by high performance anion exchange chromatography. The strains secreting cellulases outperformed the corresponding strains displaying the same cellulase on the surface in growth on PASC as the primary carbon source. The fastest growth was observed for the strain secreting Cel9A, followed by strain secreting Cel5H. By demonstrating growth of lactic acid bacteria expressing heterologous cellulases on PASC as a major carbon source for the first time, this study presents an important step towards the realization of consolidated bioprocesses involving lactic acid bacteria.

Jezik:Angleški jezik
Ključne besede:Lactococcus cremoris, surface display, lactic acid bacteria, cellulose, cellulases
Vrsta gradiva:Članek v reviji
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:FFA - Fakulteta za farmacijo
Status publikacije:Objavljeno
Različica publikacije:Objavljena publikacija
Leto izida:2025
Št. strani:13 str.
Številčenje:Vol. 438, art. 133177
PID:20.500.12556/RUL-181082 Povezava se odpre v novem oknu
UDK:60
ISSN pri članku:1873-2976
DOI:10.1016/j.biortech.2025.133177 Povezava se odpre v novem oknu
COBISS.SI-ID:246772995 Povezava se odpre v novem oknu
Datum objave v RUL:24.03.2026
Število ogledov:32
Število prenosov:11
Metapodatki:XML DC-XML DC-RDF
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Gradivo je del revije

Naslov:Bioresource technology
Založnik:Elsevier
ISSN:1873-2976
COBISS.SI-ID:4444495 Povezava se odpre v novem oknu

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

Jezik:Slovenski jezik
Ključne besede:Lactococcus cremoris, genski inženiring, celuloza, celulaza

Projekti

Financer:Republic of Slovenia, Ministry of Higher Education, Science and Innovation
Program financ.:Era-NET CoBioTech
Akronim:Cell4Chem

Financer:ANR - French National Research Agency
Številka projekta:ANR-21-COBI-0001
Naslov:Engineering microbial communities for the conversion of lignocellulose into medium-chain carboxylates
Akronim:Cell4Chem

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P4-0127
Naslov:Farmacevtska biotehnologija: znanost za zdravje

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:BI-FR/24-25-PROTEUS-001
Naslov:Inženiring in karakterizacija bakterije Lactococcus lactis z izraženimi celulazami

Financer:Campus France
Program financ.:France Excellence Scholarship – Slovenia

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