Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere

Traditionally, starting inoculants have been applied to improve ensiling of forage used for livestock feed. Here, we aimed to build up a bioinoculant composed of lactic acid-producing and lignocellulolytic bacteria (LB) derived from the Megathyrsus maximus (guinea grass) phyllosphere. For this, the...

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Autores principales:	Laura Díaz-García, Dayanne Chaparro, Hugo Jiménez, Luis Fernando Gómez-Ramírez, Adriana J. Bernal, Esteban Burbano-Erazo, Diego Javier Jiménez
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	bacterial consortia lignocellulose metagenome-assembled genomes silage bioinoculant Cellulosimicrobium Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/4338fe473c4e4a588b36a85586efcd0f
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spelling	oai:doaj.org-article:4338fe473c4e4a588b36a85586efcd0f2021-11-22T09:58:03ZTop-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere1664-302X10.3389/fmicb.2021.744075https://doaj.org/article/4338fe473c4e4a588b36a85586efcd0f2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.744075/fullhttps://doaj.org/toc/1664-302XTraditionally, starting inoculants have been applied to improve ensiling of forage used for livestock feed. Here, we aimed to build up a bioinoculant composed of lactic acid-producing and lignocellulolytic bacteria (LB) derived from the Megathyrsus maximus (guinea grass) phyllosphere. For this, the dilution-to-stimulation approach was used, including a sequential modification of the starting culture medium [Man, Rogosa, and Sharpe (MRS) broth] by addition of plant biomass (PB) and elimination of labile carbon sources. Along 10 growth-dilution steps (T1–T10), slight differences were observed in terms of bacterial diversity and composition. After the sixth subculture, the consortium started to degrade PB, decreasing its growth rate. The co-existence of Enterobacteriales (fast growers and highly abundance), Actinomycetales, Bacillales, and Lactobacillales species was observed at the end of the selection process. However, a significant structural change was noticed when the mixed consortium was cultivated in higher volume (500ml) for 8days, mainly increasing the proportion of Paenibacillaceae populations. Interestingly, Actinomycetales, Bacillales, and Lactobacillales respond positively to a pH decrease (4–5), suggesting a relevant role within a further silage process. Moreover, gene-centric metagenomic analysis showed an increase of (hemi)cellulose-degrading enzymes (HDEs) during the enrichment strategy. Reconstruction of metagenome-assembled genomes (MAGs) revealed that Paenibacillus, Cellulosimicrobium, and Sphingomonas appear as key (hemi)cellulolytic members (harboring endo-glucanases/xylanases, arabinofuranosidases, and esterases), whereas Enterococcus and Cellulosimicrobium have the potential to degrade oligosaccharides, metabolize xylose and might produce lactic acid through the phosphoketolase (PK) pathway. Based on this evidence, we conclude that our innovative top-down strategy enriched a unique bacterial consortium that could be useful in biotechnological applications, including the development/design of a synthetic bioinoculant to improve silage processes.Laura Díaz-GarcíaDayanne ChaparroHugo JiménezLuis Fernando Gómez-RamírezAdriana J. BernalEsteban Burbano-ErazoDiego Javier JiménezFrontiers Media S.A.articlebacterial consortialignocellulosemetagenome-assembled genomessilagebioinoculantCellulosimicrobiumMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	bacterial consortia lignocellulose metagenome-assembled genomes silage bioinoculant Cellulosimicrobium Microbiology QR1-502
spellingShingle	bacterial consortia lignocellulose metagenome-assembled genomes silage bioinoculant Cellulosimicrobium Microbiology QR1-502 Laura Díaz-García Dayanne Chaparro Hugo Jiménez Luis Fernando Gómez-Ramírez Adriana J. Bernal Esteban Burbano-Erazo Diego Javier Jiménez Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
description	Traditionally, starting inoculants have been applied to improve ensiling of forage used for livestock feed. Here, we aimed to build up a bioinoculant composed of lactic acid-producing and lignocellulolytic bacteria (LB) derived from the Megathyrsus maximus (guinea grass) phyllosphere. For this, the dilution-to-stimulation approach was used, including a sequential modification of the starting culture medium [Man, Rogosa, and Sharpe (MRS) broth] by addition of plant biomass (PB) and elimination of labile carbon sources. Along 10 growth-dilution steps (T1–T10), slight differences were observed in terms of bacterial diversity and composition. After the sixth subculture, the consortium started to degrade PB, decreasing its growth rate. The co-existence of Enterobacteriales (fast growers and highly abundance), Actinomycetales, Bacillales, and Lactobacillales species was observed at the end of the selection process. However, a significant structural change was noticed when the mixed consortium was cultivated in higher volume (500ml) for 8days, mainly increasing the proportion of Paenibacillaceae populations. Interestingly, Actinomycetales, Bacillales, and Lactobacillales respond positively to a pH decrease (4–5), suggesting a relevant role within a further silage process. Moreover, gene-centric metagenomic analysis showed an increase of (hemi)cellulose-degrading enzymes (HDEs) during the enrichment strategy. Reconstruction of metagenome-assembled genomes (MAGs) revealed that Paenibacillus, Cellulosimicrobium, and Sphingomonas appear as key (hemi)cellulolytic members (harboring endo-glucanases/xylanases, arabinofuranosidases, and esterases), whereas Enterococcus and Cellulosimicrobium have the potential to degrade oligosaccharides, metabolize xylose and might produce lactic acid through the phosphoketolase (PK) pathway. Based on this evidence, we conclude that our innovative top-down strategy enriched a unique bacterial consortium that could be useful in biotechnological applications, including the development/design of a synthetic bioinoculant to improve silage processes.
format	article
author	Laura Díaz-García Dayanne Chaparro Hugo Jiménez Luis Fernando Gómez-Ramírez Adriana J. Bernal Esteban Burbano-Erazo Diego Javier Jiménez
author_facet	Laura Díaz-García Dayanne Chaparro Hugo Jiménez Luis Fernando Gómez-Ramírez Adriana J. Bernal Esteban Burbano-Erazo Diego Javier Jiménez
author_sort	Laura Díaz-García
title	Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
title_short	Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
title_full	Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
title_fullStr	Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
title_full_unstemmed	Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere
title_sort	top-down enrichment strategy to co-cultivate lactic acid and lignocellulolytic bacteria from the megathyrsus maximus phyllosphere
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/4338fe473c4e4a588b36a85586efcd0f
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Top-Down Enrichment Strategy to Co-cultivate Lactic Acid and Lignocellulolytic Bacteria From the Megathyrsus maximus Phyllosphere

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