scholarly journals Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacterium Catenovulum sp. CCB-QB4

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10929
Author(s):  
Go Furusawa ◽  
Nor Azura Azami ◽  
Aik-Hong Teh
Keyword(s):  
Carbon Source ◽  
Uronic Acid ◽  
Marine Bacterium ◽  
Glycoside Hydrolases ◽  
Uronic Acids ◽  
Marine Microorganisms ◽  
Growth Experiment ◽  
Degrading Bacterium ◽  
Polysaccharide Lyases ◽  
Cazy Database

Background Oligosaccharides from polysaccharides containing uronic acids are known to have many useful bioactivities. Thus, polysaccharide lyases (PLs) and glycoside hydrolases (GHs) involved in producing the oligosaccharides have attracted interest in both medical and industrial settings. The numerous polysaccharide lyases and glycoside hydrolases involved in producing the oligosaccharides were isolated from soil and marine microorganisms. Our previous report demonstrated that an agar-degrading bacterium, Catenovulum sp. CCB-QB4, isolated from a coastal area of Penang, Malaysia, possessed 183 glycoside hydrolases and 43 polysaccharide lyases in the genome. We expected that the strain might degrade and use uronic acid-containing polysaccharides as a carbon source, indicating that the strain has a potential for a source of novel genes for degrading the polysaccharides. Methods To confirm the expectation, the QB4 cells were cultured in artificial seawater media with uronic acid-containing polysaccharides, namely alginate, pectin (and saturated galacturonate), ulvan, and gellan gum, and the growth was observed. The genes involved in degradation and utilization of uronic acid-containing polysaccharides were explored in the QB4 genome using CAZy analysis and BlastP analysis. Results The QB4 cells were capable of using these polysaccharides as a carbon source, and especially, the cells exhibited a robust growth in the presence of alginate. 28 PLs and 22 GHs related to the degradation of these polysaccharides were found in the QB4 genome based on the CAZy database. Eleven polysaccharide lyases and 16 glycoside hydrolases contained lipobox motif, indicating that these enzymes play an important role in degrading the polysaccharides. Fourteen of 28 polysaccharide lyases were classified into ulvan lyase, and the QB4 genome possessed the most abundant ulvan lyase genes in the CAZy database. Besides, genes involved in uronic acid metabolisms were also present in the genome. These results were consistent with the cell growth. In the pectin metabolic pathway, the strain had genes for three different pathways. However, the growth experiment using saturated galacturonate exhibited that the strain can only use the pathway related to unsaturated galacturonate.

scholarly journals Biochemical Reconstruction of a Metabolic Pathway from a Marine Bacterium Reveals Its Mechanism of Pectin Depolymerization

2018 ◽  
Vol 85 (1) ◽  
Author(s):  
Joanne K. Hobbs ◽  
Andrew G. Hettle ◽  
Chelsea Vickers ◽  
Alisdair B. Boraston
Keyword(s):  
Carbon Source ◽  
Marine Bacteria ◽  
Cell Walls ◽  
Marine Bacterium ◽  
Canonical Pathway ◽  
Glycoside Hydrolases ◽  
Biofuel Production ◽  
Terrestrial Plants ◽  
Metabolic Intermediate ◽  
Plant Waste

ABSTRACTPectin is a complex uronic acid-containing polysaccharide typically found in plant cell walls, though forms of pectin are also found in marine diatoms and seagrasses. Genetic loci that target pectin have recently been identified in two phyla of marine bacteria. These loci appear to encode a pectin saccharification pathway that is distinct from the canonical pathway typically associated with phytopathogenic terrestrial bacteria. However, very few components of the marine pectin metabolism pathway have been experimentally validated. Here, we biochemically reconstructed the pectin saccharification pathway from a marinePseudoalteromonassp.in vitroand show that it results in the production of galacturonate and the key metabolic intermediate 5-keto-4-deoxyuronate (DKI). We demonstrate the sequential de-esterification and depolymerization of pectin into oligosaccharides and the synergistic action of glycoside hydrolases (GHs) to fully degrade these oligosaccharides into monosaccharides. Furthermore, we show that this pathway relies on enzymes belonging to GH family 105 to carry out the equivalent chemistry afforded by an exolytic polysaccharide lyase (PL) and KdgF in the canonical pectin pathway. Finally, we synthesize our findings into a model of marine pectin degradation and compare it with the canonical pathway. Our results underline the shifting view of pectin as a solely terrestrial polysaccharide and highlight the importance of marine pectin as a carbon source for suitably adapted marine heterotrophs. This alternate pathway has the potential to be exploited in the growing field of biofuel production from plant waste.IMPORTANCEMarine polysaccharides, found in the cell walls of seaweeds and other marine macrophytes, represent a vast sink of photosynthetically fixed carbon. As such, their breakdown by marine microbes contributes significantly to global carbon cycling. Pectin is an abundant polysaccharide found in the cell walls of terrestrial plants, but it has recently been reported that some marine bacteria possess the genetic capacity to degrade it. In this study, we biochemically characterized seven key enzymes from a marine bacterium that, together, fully degrade the backbone of pectin into its constituent monosaccharides. Our findings highlight the importance of pectin as a marine carbon source available to bacteria that possess this pathway. The characterized enzymes also have the potential to be utilized in the production of biofuels from plant waste.

scholarly journals Two New Alginate Lyases of PL7 and PL6 Families from Polysaccharide-Degrading Bacterium Formosa algae KMM 3553T: Structure, Properties, and Products Analysis

Marine Drugs ◽  
2020 ◽  
Vol 18 (2) ◽  
pp. 130 ◽  
Author(s):  
Alexey Belik ◽  
Artem Silchenko ◽  
Olesya Malyarenko ◽  
Anton Rasin ◽  
Marina Kiseleva ◽  
...  
Keyword(s):  
Marine Bacterium ◽  
Tumor Therapy ◽  
Alginate Lyase ◽  
E Coli ◽  
Degrading Bacterium ◽  
Polysaccharide Lyases ◽  
Optimum Ph ◽  
Products Analysis ◽  
Alginate Lyases ◽  
Structure Properties

A bifunctional alginate lyase (ALFA3) and mannuronate-specific alginate lyase (ALFA4) genes were found in the genome of polysaccharide-degrading marine bacterium Formosa algae KMM 3553T. They were classified to PL7 and PL6 polysaccharide lyases families and expressed in E. coli. The recombinant ALFA3 appeared to be active both on mannuronate- and guluronate-enriched alginates, as well as pure sodium mannuronate. For all substrates, optimum conditions were pH 6.0 and 35 °C; Km was 0.12 ± 0.01 mg/mL, and half-inactivation time was 30 min at 42 °C. Recombinant ALFA4 was active predominately on pure sodium mannuronate, with optimum pH 8.0 and temperature 30 °C, Km was 3.01 ± 0.05 mg/mL. It was stable up to 30 °C; half-inactivation time was 1 h 40 min at 37 °C. 1H NMR analysis showed that ALFA3 degraded mannuronate and mannuronate-guluronate blocks, while ALFA4 degraded only mannuronate blocks, producing mainly disaccharides. Products of digestion of pure sodium mannuronate by ALFA3 at 200 µg/mL inhibited anchorage-independent colony formation of human melanoma cells SK-MEL-5, SK-MEL-28, and RPMI-7951 up to 17% stronger compared to native polymannuronate. This fact supports previous data and suggests that mannuronate oligosaccharides may be useful for synergic tumor therapy.

scholarly journals Genome and Secretome Analysis of Staphylotrichum longicolleum DSM105789 Cultured on Agro-Residual and Chitinous Biomass

2021 ◽  
Vol 9 (8) ◽  
pp. 1581
Author(s):  
Arslan Ali ◽  
Bernhard Ellinger ◽  
Sophie C. Brandt ◽  
Christian Betzel ◽  
Martin Rühl ◽  
...  
Keyword(s):  
Sugarcane Bagasse ◽  
Chitinase Activity ◽  
Growth Conditions ◽  
Glycoside Hydrolases ◽  
Carbohydrate Binding ◽  
Waste Products ◽  
Enzyme Mixture ◽  
Secretome Analysis ◽  
Polysaccharide Lyases ◽  
Carbohydrate Esterases

Staphylotrichum longicolleum FW57 (DSM105789) is a prolific chitinolytic fungus isolated from wood, with a chitinase activity of 0.11 ± 0.01 U/mg. We selected this strain for genome sequencing and annotation, and compiled its growth characteristics on four different chitinous substrates as well as two agro-industrial waste products. We found that the enzymatic mixture secreted by FW57 was not only able to digest pre-treated sugarcane bagasse, but also untreated sugarcane bagasse and maize leaves. The efficiency was comparable to a commercial enzymatic cocktail, highlighting the potential of the S. longicolleum enzyme mixture as an alternative pretreatment method. To further characterize the enzymes, which efficiently digested polymers such as cellulose, hemicellulose, pectin, starch, and lignin, we performed in-depth mass spectrometry-based secretome analysis using tryptic peptides from in-gel and in-solution digestions. Depending on the growth conditions, we were able to detect from 442 to 1092 proteins, which were annotated to identify from 134 to 224 putative carbohydrate-active enzymes (CAZymes) in five different families: glycoside hydrolases, auxiliary activities, carbohydrate esterases, polysaccharide lyases, glycosyl transferases, and proteins containing a carbohydrate-binding module, as well as combinations thereof. The FW57 enzyme mixture could be used to replace commercial enzyme cocktails for the digestion of agro-residual substrates.

scholarly journals A Multiomic Approach to Understand How Pleurotus eryngii Transforms Non-Woody Lignocellulosic Material

2021 ◽  
Vol 7 (6) ◽  
pp. 426
Author(s):  
Ander Peña ◽  
Rashid Babiker ◽  
Delphine Chaduli ◽  
Anna Lipzen ◽  
Mei Wang ◽  
...  
Keyword(s):  
Extracellular Enzymes ◽  
Fungal Growth ◽  
2D Nmr ◽  
Pleurotus Eryngii ◽  
Glycoside Hydrolases ◽  
Oxidative Enzymes ◽  
Initial Growth ◽  
Lignocellulosic Material ◽  
Polysaccharide Lyases ◽  
Polysaccharide Monooxygenases

Pleurotus eryngii is a grassland-inhabiting fungus of biotechnological interest due to its ability to colonize non-woody lignocellulosic material. Genomic, transcriptomic, exoproteomic, and metabolomic analyses were combined to explain the enzymatic aspects underlaying wheat–straw transformation. Up-regulated and constitutive glycoside–hydrolases, polysaccharide–lyases, and carbohydrate–esterases active on polysaccharides, laccases active on lignin, and a surprisingly high amount of constitutive/inducible aryl–alcohol oxidases (AAOs) constituted the suite of extracellular enzymes at early fungal growth. Higher enzyme diversity and abundance characterized the longer-term growth, with an array of oxidoreductases involved in depolymerization of both cellulose and lignin, which were often up-regulated since initial growth. These oxidative enzymes included lytic polysaccharide monooxygenases (LPMOs) acting on crystalline polysaccharides, cellobiose dehydrogenase involved in LPMO activation, and ligninolytic peroxidases (mainly manganese-oxidizing peroxidases), together with highly abundant H2O2-producing AAOs. Interestingly, some of the most relevant enzymes acting on polysaccharides were appended to a cellulose-binding module. This is potentially related to the non-woody habitat of P. eryngii (in contrast to the wood habitat of many basidiomycetes). Additionally, insights into the intracellular catabolism of aromatic compounds, which is a neglected area of study in lignin degradation by basidiomycetes, were also provided. The multiomic approach reveals that although non-woody decay does not result in dramatic modifications, as revealed by detailed 2D-NMR and other analyses, it implies activation of the complete set of hydrolytic and oxidative enzymes characterizing lignocellulose-decaying basidiomycetes.

scholarly journals Isolation and Characterization of a Mutant of the Marine Bacterium Alcanivorax borkumensis SK2 Defective in Lipid Biosynthesis

2010 ◽  
Vol 76 (9) ◽  
pp. 2884-2894 ◽  
Author(s):  
Efraín Manilla-Pérez ◽  
Alvin Brian Lange ◽  
Stephan Hetzler ◽  
Marc Wältermann ◽  
Rainer Kalscheuer ◽  
...  
Keyword(s):  
Carbon Source ◽  
Sole Carbon Source ◽  
Marine Bacterium ◽  
Neutral Lipids ◽  
Nile Red ◽  
Dry Weight ◽  
Wax Ester ◽  
Gene Encoding ◽  
Isolation And Characterization ◽  
Key Enzyme

ABSTRACT In many microorganisms, the key enzyme responsible for catalyzing the last step in triacylglycerol (TAG) and wax ester (WE) biosynthesis is an unspecific acyltransferase which is also referred to as wax ester synthase/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT; AtfA). The importance and function of two AtfA homologues (AtfA1 and AtfA2) in the biosynthesis of TAGs and WEs in the hydrocarbon-degrading marine bacterium Alcanivorax borkumensis SK2 have been described recently. However, after the disruption of both the AtfA1 and AtfA2 genes, reduced but substantial accumulation of TAGs was still observed, indicating the existence of an alternative TAG biosynthesis pathway. In this study, transposon-induced mutagenesis was applied to an atfA1 atfA2 double mutant to screen for A. borkumensis mutants totally defective in biosynthesis of neutral lipids in order to identify additional enzymes involved in the biosynthesis of these lipids. At the same time, we have searched for a totally TAG-negative mutant in order to study the function of TAGs in A. borkumensis. Thirteen fluorescence-negative mutants were identified on Nile red ONR7a agar plates and analyzed for their abilities to synthesize lipids. Among these, mutant 2 M131 was no longer able to synthesize and accumulate TAGs if pyruvate was used as the sole carbon source. The transposon insertion was localized in a gene encoding a putative cytochrome c family protein (ABO_1185). Growth and TAG accumulation experiments showed that the disruption of this gene resulted in the absence of TAGs in 2 M131 but that growth was not affected. In cells of A. borkumensis SK2 grown on pyruvate as the sole carbon source, TAGs represented about 11% of the dry weight of the cells, while in the mutant 2 M131, TAGs were not detected by thin-layer and gas chromatography analyses. Starvation and lipid mobilization experiments revealed that the lipids play an important role in the survival of the cells. The function of neutral lipids in A. borkumensis SK2 is discussed.

Bioavailability of a nonylphenol isomer in dependence on the association to dissolved humic substances

2004 ◽  
Vol 50 (5) ◽  
pp. 277-283 ◽  
Author(s):  
R. Vinken ◽  
A. Höllrigl-Rosta ◽  
B. Schmidt ◽  
A. Schäffer ◽  
P.F.-X. Corvini
Keyword(s):  
Humic Acid ◽  
Carbon Source ◽  
Humic Substances ◽  
Organic Compounds ◽  
Sole Carbon Source ◽  
Equilibrium Dialysis ◽  
Fulvic Acids ◽  
Degrading Bacterium ◽  
Bacterium Strain ◽  
Dissolved Humic Substances

Humic substances are important environmental components since they represent a very large part of organic compounds on earth. According to many reports, dissolved humic substances are a determinant parameter for the bioavailability of xenobiotic compounds. For the present bioavailability studies, two kinds of dissolved humic substances, a commercially available humic acid and fulvic acids isolated from peat were used. As the relevant xenobiotic, a defined branched nonylphenol isomer, 4(3′,5′-dimethyl-3′-heptyl)-phenol (p353NP) was synthesised according to Friedel-Crafts alkylation. Equilibrium dialysis studies were implemented in order to investigate the association between 14C-labelled p353NP and dissolved humic substances. The biodegradability in the presence of dissolved humic substances was examined in experiments with the nonylphenol degrading bacterium strain Sphingomonas TTNP3 and with p353NP as sole carbon source. The results showed that p353NP-humic acid associates were formed in high amounts, whereas no adducts with fulvic acids occurred. In the degradation studies with Sphingomonas TTNP3, no effects of dissolved humic substances on the bioavailability of p353NP could be observed. It was assumed that the association between nonylphenol and humic acids occurs rapidly and is reversible. Thus, the formation of "labile" complexes did not influence biodegradation rates, which were quite low.

scholarly journals Isolation and Characterization of a Novel p,p’-DDT-Degrading Bacterium: Aeromonas sp. Strain MY1 from Agricultural Soil

2020 ◽  
pp. 12-22
Author(s):  
Y. Murtala ◽  
B. C. Nwanguma ◽  
L. U. S. Ezeanyika
Keyword(s):  
Carbon Source ◽  
16S Rrna ◽  
Agricultural Soil ◽  
Sole Carbon Source ◽  
Phylogenetic Analyses ◽  
Inhibitory Effect ◽  
Aerobic Conditions ◽  
Degrading Bacterium ◽  
Isolation And Characterization

Background: Despite the banned on the use of dichlorodiphenyltrichloroethane (DDT) and other Persistent Organic Pollutants (POPs) by the Stockholm Convention for their toxicity, emerging shreds of evidence have indicated that DDT is, however, still in use in developing countries. This might increase the global burden of DDT contamination and its hazardous effects. Aim: This study focused on the isolation and characterization of p,p’-DDT-degrading bacterium from a tropical agricultural soil. Methodology: Standard isolation procedure was used for the screening and isolation of the strain. The 16S rRNA and phylogenetic analyses were used to identify the isolate and established protocols were followed to characterize the strain. Results: A new strain belonging to the genus Aeromonas was isolated from agricultural soil using minimal salt-p,p’-DDT enrichment medium. The 16S rRNA sequencing was used to identify the strain and the partial sequence was deposited in the NCBI GenBank as Aeromonas sp. Strain MY1. This mesophilic isolate was capable of utilizing up to 50 mgL-1 of p,p’-DDT as the sole carbon source at an optimum pH of 7.5 and optimum temperature of 35 °C within 120 h under aerobic conditions. Fe2+ (0.2 mgL-1) demonstrated a stimulatory effect on the p,p’-DDT degradation capacity by the strain MY1. However, Zn, Cu, Pb, Hg, Ag and Cr ions have demonstrated various patterns of inhibitory effect on the p,p’-DDT degradation capacity of the isolate at 0.2 mgL-1. The strain MY1 could be a promising candidate for the bioremediation of p,p’-DDT contaminant. Conclusion: Aeromonas sp. strain MY1 was capable of utilizing p,p’-DDT as a sole carbon source under aerobic conditions. The utilization capacity of the strain was influenced by some heavy metals. Fe was found to enhance the p,p’-DDT utilization capacity of the isolate at a lower concentration. While Zn, Cu, Pb, Hg, Ag and Cr showed various patterns of inhibitory effect.

Marine Biosurfactants, III. Toxicity Testing with Marine Microorganisms and Comparison with Synthetic Surfactants

1991 ◽  
Vol 46 (3-4) ◽  
pp. 210-216 ◽  
Author(s):  
Knut Poremba ◽  
Wilfried Gunkel ◽  
Siegmund Lang ◽  
Fritz Wagner
Keyword(s):  
Marine Bacterium ◽  
Sea Water ◽  
Toxicity Testing ◽  
Test System ◽  
Marine Microorganisms ◽  
Oil Dispersants ◽  
Test Organisms ◽  
Pure Cultures ◽  
Microtox Test ◽  
Synthetic Surfactants

Eight synthetic and nine biogenetic surfactants were tested on their toxicity. Because of their possible application as oil dispersants against oil slicks on sea. the test organisms used were marine microorganisms (mixed and pure cultures of bacteria, microalgae, and protozoa). Bacterial growth was hardly effected or stimulated, whilst that of algae and flagellates was reduced. All substances tested were biodegradaded in sea water. The bioluminescence of Photobacter phosphoreum (Microtox test) was the most sensitive test system used. A ranking shows that most biogenetic surfactants were less toxic than synthetic surfactants. No toxicity could be detected with the glucose-lipid GL. produced by the marine bacterium Alcaligenes sp. MM 1.

scholarly journals Biosynthesis of Extracellular Polymeric Substances by the Marine BacteriumSaccharophagus degradansunder Different Nutritional Conditions

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Yolanda González-García ◽  
Alejandra Heredia ◽  
Juan Carlos Meza-Contreras ◽  
Froylán M. E. Escalante ◽  
Rosa María Camacho-Ruiz ◽  
...  
Keyword(s):  
Carbon Source ◽  
Synergistic Effect ◽  
Maximum Concentration ◽  
Marine Bacterium ◽  
Extracellular Polymeric Substances ◽  
Mineral Medium ◽  
Saccharophagus Degradans ◽  
First Report ◽  
Nutritional Conditions ◽  
New Feature

The effect of carbon source, carbon to nitrogen (C/N) ratio, and limitation in nutrients (N, P, K, Ca, Mg, and Fe) on extracellular polymeric substances (EPS) synthesis by the marine bacteriumSaccharophagus degradanswas studied. This strain was able to grow in mineral medium and produce EPS with different efficiency according to the C source used (g EPS/L): glucose or starch (1.5 ± 0.2); galactose, sucrose, or xylose (0.7 ± 0.2); and fructose (0.3 ± 0.1). The C/N ratio (glucose/ammonium) had a significant effect on EPS biosynthesis due to its production rise as the C/N ratio increased from 3 to 100 (0.7 to 2.1 g EPS/L). It was also observed that limitation in nutrients such as N, P, K, Ca, Mg, and Fe also favored EPS biosynthesis. When taking into account both factors (C/N ratio, 100; nutrients limitation, 50%) a positive synergistic effect was noted on EPS production since under these conditions the maximum concentration obtained was 4.12 ± 0.3 g/L after 72 h of culture. The polymer was found to be a polysaccharide of mainly glucose, mannose, and galactose. This is the first report on EPS production byS. degradanswhich is a new feature of this versatile marine bacterium.

scholarly journals Draft Genome Sequence of Ralstonia sp. MD27, a Poly(3-Hydroxybutyrate)-Degrading Bacterium, Isolated from Compost

2015 ◽  
Vol 3 (5) ◽  
Author(s):  
Morgan Zhu ◽  
Lucy M. McCully ◽  
Mark W. Silby ◽  
Tamunonengiyeofori I. Charles-Ogan ◽  
Jean Huang ◽  
...  
Keyword(s):  
Carbon Source ◽  
Genome Sequence ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Sequence Length ◽  
Total Sequence ◽  
Degrading Bacterium ◽  
Total Sequence Length

Ralstonia sp. strain MD27, a novel biopolymer-degrading betaproteobacterium, was isolated from compost samples. This organism has been shown to utilize the biopolymer poly(3-hydroxybutyrate) [P(3HB)] as a carbon source for growth. We report the draft genome sequence of MD27 with an estimated total sequence length of 5.9 Mb.

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