Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation
Abstract Background Bacterial cellulose (BC) is a biocompatible material with unique mechanical properties, thus holding a significant industrial potential. Despite many acetic acid bacteria (AAB) being BC overproducers, cost-effective production remains a challenge. The role of pyrroloquinoline qui...
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
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| Online Access: | https://doi.org/10.1186/s13068-024-02482-9 |
| _version_ | 1797275466322673664 |
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| author | Pedro Montenegro-Silva Tom Ellis Fernando Dourado Miguel Gama Lucília Domingues |
| author_facet | Pedro Montenegro-Silva Tom Ellis Fernando Dourado Miguel Gama Lucília Domingues |
| author_sort | Pedro Montenegro-Silva |
| collection | DOAJ |
| description | Abstract Background Bacterial cellulose (BC) is a biocompatible material with unique mechanical properties, thus holding a significant industrial potential. Despite many acetic acid bacteria (AAB) being BC overproducers, cost-effective production remains a challenge. The role of pyrroloquinoline quinone (PQQ)-dependent membrane dehydrogenases (mDH) is crucial in the metabolism of AAB since it links substrate incomplete oxidation in the periplasm to energy generation. Specifically, glucose oxidation to gluconic acid substantially lowers environmental pH and hinders BC production. Conversely, ethanol supplementation is known to enhance BC yields in Komagataeibacter spp. by promoting efficient glucose utilization. Results K. sucrofermentans ATCC 700178 was engineered, knocking out the four PQQ-mDHs, to assess their impact on BC production. The strain KS003, lacking PQQ-dependent glucose dehydrogenase (PQQ-GDH), did not produce gluconic acid and exhibited a 5.77-fold increase in BC production with glucose as the sole carbon source, and a 2.26-fold increase under optimal ethanol supplementation conditions. In contrast, the strain KS004, deficient in the PQQ-dependent alcohol dehydrogenase (PQQ-ADH), showed no significant change in BC yield in the single carbon source experiment but showed a restrained benefit from ethanol supplementation. Conclusions The results underscore the critical influence of PQQ-GDH and PQQ-ADH and clarify the effect of ethanol supplementation on BC production in K. sucrofermentans ATCC 700178. This study provides a foundation for further metabolic pathway optimization, emphasizing the importance of diauxic ethanol metabolism for high BC production. |
| first_indexed | 2024-03-07T15:14:46Z |
| format | Article |
| id | doaj.art-2b3e0a2910dc4482a3b838940cf1d64f |
| institution | Directory Open Access Journal |
| issn | 2731-3654 |
| language | English |
| last_indexed | 2024-03-07T15:14:46Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj.art-2b3e0a2910dc4482a3b838940cf1d64f2024-03-05T17:57:58ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542024-02-0117111610.1186/s13068-024-02482-9Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementationPedro Montenegro-Silva0Tom Ellis1Fernando Dourado2Miguel Gama3Lucília Domingues4CEB–Center of Biological Engineering, University of MinhoCentre for Synthetic Biology, Imperial College LondonCEB–Center of Biological Engineering, University of MinhoCEB–Center of Biological Engineering, University of MinhoCEB–Center of Biological Engineering, University of MinhoAbstract Background Bacterial cellulose (BC) is a biocompatible material with unique mechanical properties, thus holding a significant industrial potential. Despite many acetic acid bacteria (AAB) being BC overproducers, cost-effective production remains a challenge. The role of pyrroloquinoline quinone (PQQ)-dependent membrane dehydrogenases (mDH) is crucial in the metabolism of AAB since it links substrate incomplete oxidation in the periplasm to energy generation. Specifically, glucose oxidation to gluconic acid substantially lowers environmental pH and hinders BC production. Conversely, ethanol supplementation is known to enhance BC yields in Komagataeibacter spp. by promoting efficient glucose utilization. Results K. sucrofermentans ATCC 700178 was engineered, knocking out the four PQQ-mDHs, to assess their impact on BC production. The strain KS003, lacking PQQ-dependent glucose dehydrogenase (PQQ-GDH), did not produce gluconic acid and exhibited a 5.77-fold increase in BC production with glucose as the sole carbon source, and a 2.26-fold increase under optimal ethanol supplementation conditions. In contrast, the strain KS004, deficient in the PQQ-dependent alcohol dehydrogenase (PQQ-ADH), showed no significant change in BC yield in the single carbon source experiment but showed a restrained benefit from ethanol supplementation. Conclusions The results underscore the critical influence of PQQ-GDH and PQQ-ADH and clarify the effect of ethanol supplementation on BC production in K. sucrofermentans ATCC 700178. This study provides a foundation for further metabolic pathway optimization, emphasizing the importance of diauxic ethanol metabolism for high BC production.https://doi.org/10.1186/s13068-024-02482-9Bacterial celluloseKomagataeibacterMetabolic engineeringAcetic acid bacteriaPQQ-dependent dehydrogenasesGluconic acid |
| spellingShingle | Pedro Montenegro-Silva Tom Ellis Fernando Dourado Miguel Gama Lucília Domingues Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation Biotechnology for Biofuels and Bioproducts Bacterial cellulose Komagataeibacter Metabolic engineering Acetic acid bacteria PQQ-dependent dehydrogenases Gluconic acid |
| title | Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation |
| title_full | Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation |
| title_fullStr | Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation |
| title_full_unstemmed | Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation |
| title_short | Enhanced bacterial cellulose production in Komagataeibacter sucrofermentans: impact of different PQQ-dependent dehydrogenase knockouts and ethanol supplementation |
| title_sort | enhanced bacterial cellulose production in komagataeibacter sucrofermentans impact of different pqq dependent dehydrogenase knockouts and ethanol supplementation |
| topic | Bacterial cellulose Komagataeibacter Metabolic engineering Acetic acid bacteria PQQ-dependent dehydrogenases Gluconic acid |
| url | https://doi.org/10.1186/s13068-024-02482-9 |
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