Degradation Of Humic Substances Research Articles

The implications of Fe speciation for the humic substance stability of ternary Fe(III)–montmorillonite–humic substance systems

Abstract Mineral–organic matter (OM) associations play an important role in determining the long-term retention of OM in soils. However, the retention mechanisms of OM in cation–mineral–OM systems remain unclear. Taking into account the dominance of montmorillonite (Mnt) in the soil of the temperate zone, we investigated the stability of humic substances (HSs) in the Fe(III)–Mnt–HS system using thermal analysis. The HS degradation started at ~387°C in the Fe(III)–Mnt–HS system, which was higher than that of the Fe(III)–HS system (290°C). The formed ferrihydrite (Fhy) mainly contributed to the enhanced labile OM retention through adsorption and/or co-precipitation, whereas Mnt inhibited the initial formation and subsequent transformation of Fhy, thus improving the stability of OM. These results suggest that the HS stability in Fe(III)–clay–HS systems depends on the Fe speciation affected by clay minerals, and this finding provides insights into OM–mineral interactions in temperate-zone soils.

Study of the effectiveness of a ZnO–TiO2 formulation in the degradation of humic substances in mature leachate by solar photocatalysis Brazil

Treatment of landfill leachate is an important environmental issue, especially in developing countries such as Brazil. Advanced oxidation processes (AOPs) have been considered interesting treatment alternatives. In this study, ZnO–TiO2 mixtures were incorporated into a paint polymer matrix and fixed onto supports. Paints were applied by overlapping coat layers on plates, resulting in high film thickness (600 ± 80 µm). Treatment of mature leachate by an AOP was conducted in a plug flow reactor connected to a stirred tank under solar irradiation. The objective was to evaluate the degradation of humic acid (HA) and fulvic acid + humins (FAH). The highest HA and FAH removal efficiencies were 62% ± 4.9% and 16% ± 4.2%, respectively. The kinetic model provided a coefficient of determination (R2) of 0.974. Rate constants for HA and FAH removal were 2.96 and 1.03 × 10−3 min−1, respectively. Statistical models for HA and FAH degradation had R2 values of 0.96 and 0.99, respectively. Both approaches indicated that HA degradation is greater at acidic pH and higher TiO2 concentrations. FAH degradation was favored by acidic pH and higher ZnO concentrations. Statistical models showed the same mean difference in conversion between replicate runs of HA and FAH, suggesting a uniform diffusion of fractions through catalysts.
 Keywords: heterogeneous photocatalysis, humic and fulvic acid, landfill leachate treatment, titanium dioxide, zinc oxide.

Soil water content as a critical factor for stable bacterial community structure and degradative activity in maritime Antarctic soil.

Recent increases in air temperature across the Antarctic Peninsula may prolong the thawing period and directly affect the soil temperature (Ts) and volumetric soil water content (SWC) in maritime tundra. Under an 8°C soil warming scenario, two customized microcosm systems with maritime Antarctic soils were incubated to investigate the differential influence of SWC on the bacterial community and degradation activity of humic substances (HS), the largest constituent of soil organic carbon and a key component of the terrestrial ecosystem. When the microcosm soil (KS1-4Feb) was incubated for 90 days (T = 90) at a constant SWC of ~32%, the initial HS content (167.0 mg/g of dried soil) decreased to 156.0 mg (approximately 6.6% loss, p < 0.05). However, when another microcosm soil (KS1-4Apr) was incubated with SWCs that gradually decreased from 37% to 9% for T = 90, HS degradation was undetected. The low HS degradative activity persisted, even after the SWC was restored to 30% with water supply for an additional T = 30. Overall bacterial community structure remained relatively stable at a constant SWC setting (KS1-4Feb). In contrast, we saw marked shifts in the bacterial community structure with the changing SWC regimen (KS1-4Apr), suggesting that the soil bacterial communities are vulnerable to drying and re-wetting conditions. These microcosm experiments provide new information regarding the effects of constant SWC and higher Ts on bacterial communities for HS degradation in maritime Antarctic tundra soil.

Surface weak acid-base pair of FeOOH/Al2O3 for enhanced peroxymonosulfate activation in degradation of humic substances from water

Three oxides (SiO2, Al2O3 and FeOOH/Al2O3) made of earth-abundant elements (O, Si, Al and Fe) were prepared to activate peroxymonosulfate (PMS). Humic acid (HA) as a representative of humic substances in water was used to evaluate their PMS activation performance. FeOOH/Al2O3 exhibited the highest catalytic activity towards HA degradation with a pseudo-first order reaction rate constant of 0.0204 min−1, but almost no HA was degraded over SiO2. The results of Electron Spin Resonance (ESR) analysis and radical quenching experiments verified that singlet oxygen (1O2) was the sole reactive species for HA degradation. The relationship between surface properties of tested oxides and their PMS activation efficiency was investigated. According to the characterization results of powder X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), nitrogen adsorption-desorption experiment, temperature programmed desorption of adsorbed CO2 (CO2-TPD) and Pyridine-FTIR spectroscopy, the surface acid-base property was proven to be responsible to excellent PMS activation performance of FeOOH/Al2O3 for 1O2 generation. A dual-reaction-site mechanism was proposed and the contribution of weak Lewis acid site and weak basic site to HA degradation in PMS-FeOOH/Al2O3 system was discussed based on all the experimental results.

Spectroscopic evidence for hyperthermophilic pretreatment intensifying humification during pig manure and rice straw composting

Hyperthermophilic pretreatment composting (HPC) is superior to traditional composting (CK) with shortened maturity period and enhanced humification degree. However, the chemical and structural evolution of humic substances (HS) at the molecular level is not known. In this study, the impact of hyperthermophilic pretreatment (90 °C, 4 h) on the content and chemical composition of HS during composting were investigated. The HS content of the final compost was 87.8 g/kg and 76.7 g/kg in HPC and CK, respectively. Significantly higher humic acid/fulvic acid ratio (1.27 in HPC v.s. 0.77 in CK) was observed in HPC. 13C NMR spectroscopic data showed a higher aromatics percentage and earlier enrichment of aromatic structures in HS extracted from HPC than CK. Intensified humification of HPC was related to the increased levels of HS precursors and degradation of lignocellulose. Redundancy analysis demonstrated that aromatic C, phenolic C and O-alkyl C can be used for evaluation of the humification degree.

The contrasting roles of aquatic fungi and oomycetes in the degradation and transformation of polymeric organic matter

AbstractStudies on the ecological role of fungi and, to a lesser extent, oomycetes, are receiving increasing attention, mainly due to their participation in the cycling of organic matter in aquatic ecosystems. To unravel their importance in humification processes, we isolated several strains of fungi and oomycetes from Anzali lagoon, Iran. We then performed taxonomic characterization by morphological and molecular methods, analyzed the ability to degrade several polymeric substrates, performed metabolic fingerprinting with Ecoplates, and determined the degradation of humic substances (HS) using liquid chromatography‐organic carbon detection. Our analyses highlighted the capacity of aquatic fungi to better degrade a plethora of organic molecules, including complex polymers. Specifically, we were able to demonstrate not only the utilization of these complex polymers, but also the role of fungi in the production of HS. In contrast, oomycetes, despite some morphological and physiological similarities with aquatic fungi, exhibited a propensity toward opportunism, quickly benefitting from the availability of small organic molecules, while exhibiting sensitivity toward more complex polymers. Despite their contrasting roles, our study highlights the importance of both oomycetes and fungi in aquatic organic matter transformation and cycling with potential implications for the global carbon cycle.

Improved dewaterability of anaerobically digested sludge and compositional changes in extracellular polymeric substances by indigenous persulfate activation

In this study, an indigenous activation of persulfate by iron-bearing minerals was examined using peroxymonosulfate (PMS) and peroxydisulfate (PDS) for the dewaterability and extracellular polymeric substances (EPS) composition of anaerobically digested sludge (ADS). Iron minerals originally present in the ADS seemed to operate as an initiator of the persulfate activation, which was indicated by an increase in the total dissolved iron in the supernatant. The PMS showed higher performance in improving the ADS dewaterability compared to the PDS, with more reduction in the average microbial flocs size. The extracted EPS consisted of three different fluorescent components of tryptophan-like (C1), humic-like (C2), and fulvic-like (C3) components. In the tightly bound (TB)-EPS, two humic-like components (C2 and C3), which were resistant to degradation, exhibited a strong linkage with dewaterability; whereas, in both the PMS and the PDS systems, the C1 was preferentially degraded by the radical-based oxidation, with a greater extent for the PMS-based treatments. The results showed that prior to the actual EPS degradation, the produced radicals were initially involved in cell disruption. The SEC results during the oxidation clearly demonstrated the degradation of a large-sized biopolymer fraction, followed by the production of relatively small sized molecules. They also revealed in the TB-EPS a close association between the ADS dewaterability and the degradation of humic substances.

Pre-coagulation coupled with sponge-membrane filtration for organic matter removal and membrane fouling control during drinking water treatment

A new hybrid system was developed in this study for the treatment of drinking water consisting of pre-coagulation using polyaluminium chloride (PACl) and membrane filtration (MF) with sponge cubes acting as biomass carriers (P-SMF). When compared to a conventional MF (CMF) and a MF after coagulation by utilizing PACl (P-MF), better removal of nutrients, UV254 and dissolved organic carbon (DOC) (>65%) was obtained from the P-SMF. The accumulation of biopolymers (including polysaccharides and proteins), humic substances, hydrophilic organics, and other small molecular weight (MW) organic matter in the CMF led to the most severe membrane fouling coupled with the highest pore blocking and cake resistance. Pre-coagulation was ineffective in eliminating small MW and hydrophilic organic matter. Conversely, the larger MW organics (i.e. biopolymers and humic substances), small MW organics and hydrophilic organic compounds could be removed in significantly larger quantities in the P-SMF by PACl coagulation. This was achieved via adsorption and the biodegradation by attached biomass on these sponges and by the suspended sludge. Further analyses of the microbial community indicated that the combined addition of PACl and sponges generated a high enrichment of Zoolgloea, Amaricoccus and Reyranella leading to the reduction of biopolymers, and Flexibacter and Sphingobium were linked to the degradation of humic substances. Moreover, some members of Alphaproteobacteria in the P-SMF may be responsible for the removal of low MW organics. These results suggest that the pre-coagulation process coupled with adding sponge in the MF system is a promising technology for mitigating membrane fouling.

Biodegradation of humic substances by microscopic filamentous fungi: chromatographic and spectroscopic proxies

The study of interactions between humic substances (HSs) and soil filamentous fungi is the key to understanding the sustainable soil functioning. The present work aims to examine the decomposition of HSs by filamentous dark-pigmented fungus Alternaria alternatа under the laboratory conditions and to determine the effect of easily assimilable organic carbon on this process. Analyzing such polydisperse substances like HSs by a complex integrated methodology makes it possible to explore the data on their decomposition by microorganisms. To achieve the aforementioned goals, we used chromatographic and spectroscopic approaches: low-pressure size-exclusion and hydrophobic interaction chromatography accompanied by absorption and fluorescence spectroscopy. To determine the effect cometabolism conditions produced on HS decomposition, two types of carbon substrates were added to the nutrient media: easily assimilable organic carbon (standard 0.3% or reduced 0.03% sucrose content) and hardly assimilable organic carbon (HSs), as well as their combinations. Five HS samples of different organic matter origin have been inspected: potassium humates (HPs) and humic acids (HAs) from coal, peat, and lignosulfonate. Correlation matrix and principal component analysis (PCA) were calculated for comprehensive data analysis. Transformations of the investigated HSs under fungal cultivation lead to the increase in the low molecular weight fraction, rise of hydrophilic fraction, enlargement of absorbance ratio A250/A365, shortening of the emission wavelength of the humic-type fluorescence, and growth in the fluorescence quantum yield measured with excitation at 355 nm. A positive correlation was observed between the accumulation of fungal biomass and the degree of HS decomposition. PCA analysis confirms that the difference in the results of HS decomposition largely depends on the sucrose content and the nature of HSs. We divided all the HS samples into four groups according to the degree of HS decomposition: original HS solutions, HPs altered using fungal cultivation at 0.03% sucrose, HAs after fungal cultivation at 0.03% sucrose, and finally, HSs (both HPs and HAs) after fungal cultivation at 0.3% sucrose. In the laboratory experiments, we showed that (1) the isolated HAs were more effectively degraded than the parent HPs, and this process was more pronounced at a reduced sucrose content, and (2) the decomposition of stable organic compounds (HSs) was activated by the easily assimilable carbon sources (especially 0.3% sucrose) being present. We assume that it is the easily assimilable organic carbon that most likely triggers the HS degradation working as the priming effect in natural environments.

Laccase Production and Humic Acids Decomposition by Microscopic Soil Fungi

Laccase (para-diphenol:oxygen oxidoreductase, EC 1.10.3.2) is a phenol oxidase widespread in fungi and bacteria. In basidiomycetes, this enzyme is involved in the transformation of lignin and humic substances (HS) in soil. The role of laccases of soil ascomycetes and deuteromycetes in HS degradation is not established, and conditions of the enzyme production have been poorly studied. In the present work soil micromycetes, potential laccase producers, were isolated from typical soils of the forest, steppe, and foreststeppe zones of European Russia by plating on agar media with ABTS (2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulphonic acid, sodium salt)) as the substrate. Their abundance, species composition, conditions of laccase production, and its relation to humic acids (HA) degradation in liquid and solid media were studied. Out of 68 strains isolated, 20 exhibited ABTS oxidation at initial plating on agar media. In pure cultures on agar media, oxidation was less pronounced, but in the presence of HA laccase production by some strains was higher than without HA. Significant and weak extracellular laccase production in liquid medium was observed for Acremonium murorum (Corda) W. Gams Z1710 and Botritis cinerea Pers. ex Fries Z1711, respectively. The level of laccase production by A. murorum was the same without inducers and in the presence of HA, while B. cinerea produced laccase only without inducers. No direct correlation was found between the presence of laccase and/or its activity and ability of the fungi to decolorize (degrade) HA. In liquid media active laccase producer A. murorum caused lower HA decolorization (43%) than B. cinerea (62%) and the fungi lacking extracellular laccase (54–81%). The role of micromycete oxidative systems in HA degradation requires further investigation.

Sonocatalytic Degradation of Humic Substances From Aquatic Environments Using MgO Nanoparticles

Humic substances are considered as one of the major natural organic contaminants in water resources. Presence of such substances in the drinking water supply resources results in poor water quality and jeopardizes the consumer’s health. In the present study, sonocatalytic decomposition of the humic substances by using MgO nanoparticles was investigated under the radiation of the ultrasonic waves. Accordingly, the X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques were used to determine characteristics of the nanoparticles. Furthermore, the effects of various parameters, including the amount of nanoparticles, initial pH, initial concentration of humic substances, contact time, as well as the power and frequency of ultrasonic waves on the efficiency of the sonocatalytic decomposition of humic substances were investigated. Results of the present study showed that the efficiency of sonocatalytic decomposition of humic substances was reduced by increasing the initial pH and initial concentration of the humic substances; while, the increased amount of the nanocatalyst resulted in the increased efficiency of decomposition. Furthermore, the obtained results indicated that the MgO sonocatalytic process could decompose the humic substances with the efficiency of 78.5% under optimal conditions (pH:7, initial concentration of humic substances: 20 mg/L, concentration of nanoparticles:1 g/L, power: 100%, frequency: 37 kHz and contact time:120 min). Based on the obtained results, the sonocatalytic method using MgO can remove humic substances and other similar organic matters from the aquatic environments with a high efficiency.

Increased Electron-Accepting and Decreased Electron-Donating Capacities of Soil Humic Substances in Response to Increasing Temperature.

The electron transfer capacities (ETCs) of soil humic substances (HSs) are linked to the type and abundance of redox-active functional moieties in their structure. Natural temperature can affect the chemical structure of natural organic matter by regulating their oxidative transformation and degradation in soil. However, it is unclear if there is a direct correlation between ETC of soil HS and mean annual temperature. In this study, we assess the response of the electron-accepting and -donating capacities (EAC and EDC) of soil HSs to temperature by analyzing HSs extracted from soil set along glacial-interglacial cycles through loess-palaeosol sequences and along natural temperature gradients through latitude and altitude transects. We show that the EAC and EDC of soil HSs increase and decrease, respectively, with increasing temperature. Increased temperature facilitates the prevalence of oxidative degradation and transformation of HS in soils, thus potentially promoting the preferentially oxidative degradation of phenol moieties of HS or the oxidative transformation of electron-donating phenol moieties to electron-accepting quinone moieties in the HS structure. Consequently, the EAC and EDC of HSs in soil increase and decrease, respectively. The results of this study could help to understand biogeochemical processes, wherein the redox functionality of soil organic matter is involved in the context of increasing temperature.

Photocatalytic degradation of humic substances in the presence of ZnO nanoparticles immobilized on glass plates under ultraviolet irradiation

ABSTRACTZnO nanoparticles were prepared through the chemical precipitation method and immobilized on glass plates. The effects of initial pH, UV lamp power, glass surface area, and initial humic substances (HS) concentration on the photocatalysis of HS were studied. Acidic conditions favoured the photocatalysis of HS. The degradation efficiency of HS was enhanced by increasing the UV lamp power and glass surface area. According to the results of total organic carbon (TOC) analysis, a mineralization efficiency of 59.3% was obtained within a reaction time of 120 min in comparison with the removal efficiency (%) of UV254, which was found to be 83.3%.

Changes in Structure and Content Humic Substances in Soil During the Laboratory Simulated Fires

Abstract The paper deals with the process of thermal degradation of humic substances in soil samples exposed to increased temperature. To determine the basic properties of humic substances, humic and fulvic acids are used conventional fractionation chemical laboratory methods. To determine changes in the chemical structure, the method of use of FT-IR ATR spectroscopy technique.

Application of experimental design approach for optimization of the photocatalytic degradation of humic substances in aqueous solution using immobilized ZnO nanoparticles

Degradation of humic substances in water is important due to its adverse effects on the environment and human health. The aim of this study was modeling and investigating the degradation of humic substances in water using immobilized ZnO as a catalyst. ZnO nanoparticles were synthesized through simple coprecipitation (CPT) method and immobilized on glass plates. The immobilized ZnO nanocatalyst was characterized through scanning electron microscopy (SEM) and X-ray diffraction (XRD). Response surface methodology (RSM) and central composite design (CCD) were used to create an experimental design for humic degradation and color removal efficiency. The most important parameters including initial concentration, pH, and contact time were optimized. The optimum conditions were initial concentration of 7.68 mg/l, pH of 4.42, and contact time of about 125.6 minutes. Under optimal conditions, maximum humic substances and color removal of about 100 and 82.37% were obtained, respectively. These results illustrate that an immobilized form of ZnO can be used as an efficient nanocatalyst for effective degradation of humic substances in water.

Photocatalytic degradation of humic substances in aqueous solution using Cu-doped ZnO nanoparticles under natural sunlight irradiation.

In this study, Cu-doped ZnO nanoparticles were investigated as an efficient synthesized catalyst for photodegradation of humic substances in aqueous solution under natural sunlight irradiation. Cu-doped ZnO nanocatalyst was prepared through mild hydrothermal method and was characterized using FT-IR, powder XRD and SEM techniques. The effect of operating parameters such as doping ratio, initial pH, catalyst dosage, initial concentrations of humic substances and sunlight illuminance were studied on humic substances degradation efficiency. The results of characterization analyses of samples confirmed the proper synthesis of Cu-doped ZnO nanocatalyst. The experimental results indicated the highest degradation efficiency of HS (99.2%) observed using 1.5% Cu-doped ZnO nanoparticles at reaction time of 120 min. Photocatalytic degradation efficiency of HS in a neutral and acidic pH was much higher than that at alkaline pH. Photocatalytic degradation of HS was enhanced with increasing the catalyst dosage and sunlight illuminance, while increasing the initial HS concentration led to decrease in the degradation efficiency of HS. Conclusively, Cu-doped ZnO nanoparticles can be used as a promising and efficient catalyst for degradation of HS under natural sunlight irradiation.

Temporal changes in soil bacterial diversity and humic substances degradation in subarctic tundra soil.

Humic substances (HS), primarily humic acids (HA) and fulvic acids (FA), are the largest constituent of soil organic matter. In microcosm systems with subarctic HS-rich tundra soil (site AK 1-75; approximately 5.6 °C during the thawing period) from Council, Alaska, the HA content significantly decreased to 48% after a 99-day incubation at 5 °C as part of a biologically mediated process. Accordingly, levels of FA, a putative byproduct of HA degradation, consistently increased to 172% during an identical incubation process. Culture-independent microbial community analysis showed that during the microcosm experiments, the relative abundance of phyla Proteobacteria (bacteria) and Euryarchaeota (archaea) largely increased, indicating their involvement in HS degradation. When the indigenous bacteria in AK 1-75 were enriched in an artificial mineral medium spiked with HA, the changes in relative abundance were most conspicuous in Proteobacteria (from 60.2 to 79.0%), specifically Betaproteobacteria-related bacteria. One hundred twenty-two HA-degrading bacterial strains, primarily from the genera Paenibacillus (phylum Firmicutes) and Pseudomonas (class Gammaproteobacteria), were cultivated from AK 1-75 and nearby sites. Through culture-dependent analysis with these bacterial isolates, we observed increasing HS-degradation rates in parallel with rising temperatures in a range of 0 °C to 20 °C, with the most notable increase occurring at 8 °C compared to 6 °C. Our results indicate that, although microbial-mediated HS degradation occurs at temperature as low as 5 °C in tundra ecosystems, increasing soil temperature caused by global climate change could enhance HS degradation rates. Extending the thawing period could also increase degradation activity, thereby directly affecting nearby microbial communities and rhizosphere environments.

Humic substances enhance growth and respiration in the basidiomycetes Trametes maxima under carbon limited conditions.

Humic substances (HS) represent the major reservoir of carbon (C) in ecosystems, and their turnover is crucial for understanding the global C cycle. Although basidiomycetes clearly have a role in HS degradation, much less is known about the effect of HS on fungal traits. We studied the alteration of physiological, biochemical, and morphological characteristics of Trametes maxima in the presence of HS. Both complete medium and minimal (C-limited) medium mimicking natural environmental conditions were used. Adding HS led to increased biomass yield, but under C-limited conditions the effect was more apparent. This result indicated that HS were used as an additional substrate and agreed with data showing a greater penetration of tritium-labeled HS into the cell interior under C-limited conditions. Humic substances induced ultra-structural changes in fungal cells, especially under C limitation, including reducing the thicknesses of the hyphal sheath and cell wall. In the minimal medium, cellular respiration increased nearly three-fold under HS application, while the corresponding effect in complete medium was lower. In addition, in the presence of inhibitors, HS stimulated either the cytochrome or the alternative pathway of respiration, depending on presence or absence of glucose in the medium. Our results suggest that, under conditions mimicking the natural environment, HS may play three major roles: as a surplus substrate for fungal growth, as a factor positively affecting cell morphology, and as an activator of physiological respiration.

Versatile peroxidase degradation of humic substances: Use of isothermal titration calorimetry to assess kinetics, and applications to industrial wastes

The kinetic constants of a hybrid versatile-peroxidase (VP) which oxidizes complex polymeric humic substances (HS) derived from lignin (humic and fulvic acids) and industrial wastes were determined for the first time using isothermal titration calorimetry (iTC). The reaction conditions were manipulated to enable manganese-peroxidase (MnP) and/or lignin-peroxidase (LiP) activities to be evaluated. The peroxidase reactions exhibited varying degrees of product inhibition or activation; properties which have not previously been reported for VP enzymes. In contrast to previous work (Ertan et al., 2012) on small non-polymeric substrates (MnSO4, veratryl alcohol and dyes), all kinetic plots for polymeric HS were sigmoidal, lacked Michaelis–Menten characteristics, and were indicative of positive cooperativity. Under conditions when both LiP and MnP were active, the kinetic data fitted to a novel biphasic Hill Equation, and the rate of enzymatic reaction was significantly greater than the sum of individual LiP plus MnP activities implying synergistic activation. By employing size-exclusion chromatography and electrospray ionization mass spectrometry, the characteristics of the oxidative degradation products of the HS were also monitored. Our study showed that the allosteric behaviour of the VP enzyme promotes a high level of regulation of activity during the breakdown of model and industrial ligninolytic substrates. The work was extended to examine the kinetics of breakdown of industrial wastes (effluent from a pulp and paper plant, and fouled membrane solids extracted from a ground water treatment membrane) revealing unique, VP-mediated, kinetic responses. This work demonstrates that iTC can be successfully employed to study the kinetic properties of VP enzymes in order to devise reaction conditions optimized for oxidative degradation of HS present in materials used in a wide range of industries.

Isolation and characterization of humic substances‐degrading bacteria from the subarctic Alaska grasslands

Humic substances (HS), an important fraction of soil organic carbon, are distributed widely throughout cold environments. A total of cold-adapted 122 bacterial strains were isolated from 66 Alaska grassland soil samples based on their ability to grow on humic acids (HA), a main fraction of HS, as a carbon and energy source. These isolates were identified based on 16S rRNA gene sequencing, with class Bacilli (79.5%) and γ-Proteobacteria (17.1%) comprising the largest groups. Among them, 45 strains, mainly Paenibacillus (27 strains) and Pseudomonas (15 strains), were selected for further screening. Two strains (Pseudomonas sp. PAMC 26793 and Paenibacillus sp. PAMC 26794) most efficiently degraded HA, but showed significant differences in their ability to grow on various monocyclic aromatics, which are putative degradative metabolites of HS. Fourier transform infrared spectra also showed substantial but different changes in HA chemical structure after incubation with each strain. Gel permeation chromatography demonstrated that depolymerization and polymerization of HA occurred during HS degradation by these newly isolated microbes.