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

2021 ◽  
Vol 12 ◽  
Author(s):  
Laura Díaz-García ◽  
Dayanne Chaparro ◽  
Hugo Jiménez ◽  
Luis Fernando Gómez-Ramírez ◽  
Adriana J. Bernal ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Plant Biomass ◽  
Selection Process ◽  
Carbon Sources ◽  
Bacterial Consortium ◽  
Top Down ◽  
Relevant Role ◽  
Ph Decrease ◽  
Silage Process ◽  
Enrichment Strategy

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.

scholarly journals Dilution-to-Stimulation/Extinction Method: a Combination Enrichment Strategy To Develop a Minimal and Versatile Lignocellulolytic Bacterial Consortium

2020 ◽  
Vol 87 (2) ◽  
Author(s):  
Laura Díaz-García ◽  
Sixing Huang ◽  
Cathrin Spröer ◽  
Rocío Sierra-Ramírez ◽  
Boyke Bunk ◽  
...  
Keyword(s):  
Microbial Consortium ◽  
Plant Biomass ◽  
Bacterial Species ◽  
Bacterial Consortium ◽  
Priority Effects ◽  
Plant Residues ◽  
Agricultural Plant ◽  
Enrichment Strategy ◽  
Sequence Types ◽  
Bacterial Sequence

ABSTRACT The engineering of complex communities can be a successful path to understand the ecology of microbial systems and improve biotechnological processes. Here, we developed a strategy to assemble a minimal and effective lignocellulolytic microbial consortium (MELMC) using a sequential combination of dilution-to-stimulation and dilution-to-extinction approaches. The consortium was retrieved from Andean forest soil and selected through incubation in liquid medium with a mixture of three types of agricultural plant residues. After the dilution-to-stimulation phase, approximately 50 bacterial sequence types, mostly belonging to the Sphingobacteriaceae, Enterobacteriaceae, Pseudomonadaceae, and Paenibacillaceae, were significantly enriched. The dilution-to-extinction method demonstrated that only eight of the bacterial sequence types were necessary to maintain microbial growth and plant biomass consumption. After subsequent stabilization, only two bacterial species (Pseudomonas sp. and Paenibacillus sp.) became highly abundant (>99%) within the MELMC, indicating that these are the key players in degradation. Differences in the composition of bacterial communities between biological replicates indicated that selection, sampling, and/or priority effects could shape the consortium structure. The MELMC can degrade up to ∼13% of corn stover, consuming mostly its (hemi)cellulosic fraction. Tests with chromogenic substrates showed that the MELMC secretes an array of endoenzymes able to degrade xylan, arabinoxylan, carboxymethyl cellulose, and wheat straw. Additionally, the metagenomic profile inferred from the phylogenetic composition along with an analysis of carbohydrate-active enzymes of 20 bacterial genomes support the potential of the MELMC to deconstruct plant polysaccharides. This capacity was mainly attributed to the presence of Paenibacillus sp. IMPORTANCE The significance of our study mainly lies in the development of a combined top-down enrichment strategy (i.e., dilution to stimulation coupled to dilution to extinction) to build a minimal and versatile lignocellulolytic microbial consortium. We demonstrated that mainly two selectively enriched bacterial species (Pseudomonas sp. and Paenibacillus sp.) are required to drive the effective degradation of plant polymers. Our findings can guide the design of a synthetic bacterial consortium that could improve saccharification (i.e., the release of sugars from agricultural plant residues) processes in biorefineries. In addition, they can help to expand our ecological understanding of plant biomass degradation in enriched bacterial systems.

scholarly journals An investigation on the degradation behaviors of Mg wires/PLA composite for bone fixation implants: influence of wire content and load mode

2021 ◽  
Vol 28 (1) ◽  
pp. 39-47
Author(s):  
Xuan Li ◽  
Yu Cong ◽  
Jisheng Sui ◽  
Xiaolong Li
Keyword(s):  
Lactic Acid ◽  
Dynamic Load ◽  
Degradation Rate ◽  
Convective Transport ◽  
Poly Lactic Acid ◽  
Mechanical Loss ◽  
Bone Fixation ◽  
Load Mode ◽  
Degradation Behaviors ◽  
Ph Decrease

Abstract Poly-lactic acid based biocomposite strengthened with magnesium alloy wires (Mg wires/PLA composite) is prepared for bone fixation implantation. The influence of wire content and load mode on the degradation performances of the composite and its components is studied. The result suggests the degradation of Mg wires could slow down the pH decrease originated from the degradation of PLA, while a relatively high wire content contributes to descend the degradation rate of Mg wire in the composite. Dynamic load significantly promotes the mechanical loss of the specimens. After 30 days immersion, the Sb retention is about 65%, 52% and 55%, respectively for pure PLA, the composite at 10 vol% and 20 vol% under dynamic load, comparing to 75%, 70% and 72% under no load. Moreover, dynamic load could further mitigate the degradation of Mg wires by increasing convective transport of acidic products out of the composite.

scholarly journals Screening and directed evolution of transporters to improve xylodextrin utilization in the yeast Saccharomyces cerevisiae

2017 ◽  
Author(s):  
Chenlu Zhang ◽  
Ligia Acosta-Sampson ◽  
Vivian Yaci Yu ◽  
Jamie H. D. Cate
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Directed Evolution ◽  
Plant Biomass ◽  
Carbon Sources ◽  
Industrial Applications ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cellulosic Biofuel ◽  
Economic Production ◽  
Report Analysis ◽  
Selection Medium

AbstractThe economic production of cellulosic biofuel requires efficient and full utilization of all abundant carbohydrates naturally released from plant biomass by enzyme cocktails. Recently, we reconstituted the Neurospora crassa xylodextrin transport and consumption system in Saccharomyces cerevisiae, enabling growth of yeast on xylodextrins aerobically. However, the consumption rate of xylodextrin requires improvement for industrial applications, including consumption in anaerobic conditions. As a first step in this improvement, we report analysis of orthologues of the N. crassa transporters CDT-1 and CDT-2. Transporter ST16 from Trichoderma virens enables faster aerobic growth of S. cerevisiae on xylodextrins compared to CDT-2. ST16 is a xylodextrin-specific transporter, and the xylobiose transport activity of ST16 is not inhibited by cellobiose. Other transporters identified in the screen also enable growth on xylodextrins including xylotriose. Taken together, these results indicate that multiple transporters might prove useful to improve xylodextrin utilization in S. cerevisiae. Efforts to use directed evolution to improve ST16 from a chromosomally-integrated copy were not successful, due to background growth of yeast on other carbon sources present in the selection medium. Future experiments will require increasing the baseline growth rate of the yeast population on xylodextrins, to ensure that the selective pressure exerted on xylodextrin transport can lead to isolation of improved xylodextrin transporters.

Trust, but verify.. Power relations and control practices in a smart working environment

2021 ◽  
pp. 24-50
Author(s):  
Silvia Doria
Keyword(s):  
Power Relations ◽  
New Technologies ◽  
Working Environment ◽  
Top Down ◽  
New Ways Of Working ◽  
The World ◽  
Relevant Role ◽  
Smart Working ◽  
Banking Institution ◽  
And Control

The world of working is changing and the technological transformations are playing a relevant role in this change. In particular, new technologies are making the physical boundaries of traditional offices increasingly permeable, allowing the diffusion of New Ways of Working (Demerouti et al., 2014; Koops and Helms, 2014), such as smart working. This paper, based on a qualitative research and discursive interviews, intends to reflect on the introduction and top-down management of smart working within a banking institution. At the same time, it aims to grasp the role attributed to and played by technology in its implementation. Starting from the two reconstructed stories, I shall show if and how the innovations introduced whereby technologies enable us to work remotely, are changing existing power relations and what control dynamics emerge from the field.

scholarly journals ABC Transporters Required for Hexose Uptake byClostridium phytofermentans

2019 ◽  
Vol 201 (15) ◽  
Author(s):  
Tristan Cerisy ◽  
Alba Iglesias ◽  
William Rostain ◽  
Magali Boutard ◽  
Christine Pelle ◽  
...  
Keyword(s):  
Abc Transporters ◽  
Extracellular Enzymes ◽  
Plant Biomass ◽  
Carbon Sources ◽  
Anaerobic Bacteria ◽  
Model Organism ◽  
Industrial Transformation ◽  
Content Type ◽  
Plant Matter

ABSTRACTThe mechanisms by which bacteria uptake solutes across the cell membrane broadly impact their cellular energetics. Here, we use functional genomic, genetic, and biophysical approaches to reveal howClostridium(Lachnoclostridium)phytofermentans, a model bacterium that ferments lignocellulosic biomass, uptakes plant hexoses using highly specific, nonredundant ATP-binding cassette (ABC) transporters. We analyze the transcription patterns of its 173 annotated sugar transporter genes to find those upregulated on specific carbon sources. Inactivation of these genes reveals that individual ABC transporters are required for uptake of hexoses and hexo-oligosaccharides and that distinct ABC transporters are used for oligosaccharides versus their constituent monomers. The thermodynamics of sugar binding shows that substrate specificity of these transporters is encoded by the extracellular solute-binding subunit. As sugars are not phosphorylated during ABC transport, we identify intracellular hexokinases based onin vitroactivities. These mechanisms used byClostridiato uptake plant hexoses are key to understanding soil and intestinal microbiomes and to engineer strains for industrial transformation of lignocellulose.IMPORTANCEPlant-fermentingClostridiaare anaerobic bacteria that recycle plant matter in soil and promote human health by fermenting dietary fiber in the intestine.Clostridiadegrade plant biomass using extracellular enzymes and then uptake the liberated sugars for fermentation. The main sugars in plant biomass are hexoses, and here, we identify how hexoses are taken in to the cell by the model organismClostridium phytofermentans. We show that this bacterium uptakes hexoses using a set of highly specific, nonredundant ABC transporters. Once in the cell, the hexoses are phosphorylated by intracellular hexokinases. This study provides insight into the functioning of abundant members of soil and intestinal microbiomes and identifies gene targets to engineer strains for industrial lignocellulosic fermentation.

scholarly journals The regulatory and transcriptional landscape associated with carbon utilization in a filamentous fungus

2020 ◽  
Vol 117 (11) ◽  
pp. 6003-6013 ◽  
Author(s):  
Vincent W. Wu ◽  
Nils Thieme ◽  
Lori B. Huberman ◽  
Axel Dietschmann ◽  
David J. Kowbel ◽  
...  
Keyword(s):  
Cell Wall ◽  
Transcription Factors ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Plant Biomass ◽  
Carbon Sources ◽  
Cell Wall Degrading Enzymes ◽  
Degrading Enzymes ◽  
Genes Encoding

Filamentous fungi, such asNeurospora crassa, are very efficient in deconstructing plant biomass by the secretion of an arsenal of plant cell wall-degrading enzymes, by remodeling metabolism to accommodate production of secreted enzymes, and by enabling transport and intracellular utilization of plant biomass components. Although a number of enzymes and transcriptional regulators involved in plant biomass utilization have been identified, how filamentous fungi sense and integrate nutritional information encoded in the plant cell wall into a regulatory hierarchy for optimal utilization of complex carbon sources is not understood. Here, we performed transcriptional profiling ofN. crassaon 40 different carbon sources, including plant biomass, to provide data on how fungi sense simple to complex carbohydrates. From these data, we identified regulatory factors inN. crassaand characterized one (PDR-2) associated with pectin utilization and one with pectin/hemicellulose utilization (ARA-1). Using in vitro DNA affinity purification sequencing (DAP-seq), we identified direct targets of transcription factors involved in regulating genes encoding plant cell wall-degrading enzymes. In particular, our data clarified the role of the transcription factor VIB-1 in the regulation of genes encoding plant cell wall-degrading enzymes and nutrient scavenging and revealed a major role of the carbon catabolite repressor CRE-1 in regulating the expression of major facilitator transporter genes. These data contribute to a more complete understanding of cross talk between transcription factors and their target genes, which are involved in regulating nutrient sensing and plant biomass utilization on a global level.

scholarly journals Biodegradation Potential and Diversity of Diclofenac-degrading Microbiota in an Immobilized Cell Biofilter

Processes ◽  
2019 ◽  
Vol 7 (9) ◽  
pp. 554 ◽  
Author(s):  
Efstathia Navrozidou ◽  
Nikolaos Remmas ◽  
Paraschos Melidis ◽  
Dimitrios G. Karpouzas ◽  
George Tsiamis ◽  
...  
Keyword(s):  
Activated Sludge ◽  
High Throughput Sequencing ◽  
Rhodopseudomonas Palustris ◽  
Carbon Sources ◽  
High Strength ◽  
Adaptation Period ◽  
Immobilized Cell ◽  
Wide Range ◽  
Acidic Conditions ◽  
Ph Decrease

Despite that diclofenac has been embodied to the European watch list of priority substances of concern, studies on diclofenac biodegradation are limited and the diversity of diclofenac-degrading microbiota remains unknown. In this work, an immobilized cell biofilter was constructed and operated to evaluate its effectiveness to depurate high strength diclofenac wastewater and to identify the diclofenac-degrading community accommodated in activated sludge by employing high-throughput sequencing techniques. After a two-month adaptation period, biofilter removal efficiencies reached values as high as 97.63 ± 0.62%, whereas utilization of diclofenac in the immobilized cell biofilter led to a drastic pH decrease. Based on Illumina sequencing, the major bacterial taxa identified in the immobilized cell biofilter were members of the species Granulicella pectinivorans and Rhodanobacter terrae, followed by members of the species Castellaniella denitrificans, Parvibaculum lavamentivorans, Bordetella petrii, Bryocella elongata and Rhodopseudomonas palustris. The ability of such taxa to utilize a wide range of carbon sources and to effectively adapt under acidic conditions seemed to be the main parameters, which favored their prevalence in the immobilized cell biofilter. In addition, Wickerhamiella was the predominant fungal taxon in the immobilized cell biofilter, which appears to be actively involved in diclofenac degradation in activated sludge systems.

Histamine and Tyramine Production by a Lactobacillus Strain Subjected to External pH Decrease

1994 ◽  
Vol 57 (3) ◽  
pp. 259-262 ◽  
Author(s):  
RIITTA MAIJALA
Keyword(s):  
Lactic Acid ◽  
Room Temperature ◽  
Lactobacillus Strain ◽  
Ph Values ◽  
Initial Ph ◽  
Ph Decrease ◽  
In The Beginning ◽  
Influence Of Ph ◽  
External Ph ◽  
Made In

The preservative effect of fermentation is based on the pH decrease during processing. However, most studies conceming the influence of pH on the formation of biogenic amines have been made in broths with different initial pH values. A histamine- and tyramine-positive Lactobacillus strain isolated from dry sausage was added at an initial level of 3.9- to 4.4-logl0 CFU/ml and incubated on a shaker at room temperature (20.0°C) for 6 days. The pH was decreased in histidine- or tyrosine-fortified MRS broth by adding glucono-delta-lactone (GDL) or lactic acid during the incubation. The external acidification decreased the growth and the production of histamine and tyramine by the strain. GDL was more effective as a preventative than lactic acid. According to these results, a rapid pH decrease resulting in decreased growth of amine-positive lactic acid bacteria in the beginning of fermentation may be a means of preventing the formation of high levels of amines in foods.

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