An efficient biosurfactant-producing and crude-oil emulsifying bacterium Bacillus methylotrophicus USTBa isolated from petroleum reservoir

An efficient biosurfactant-producing bacterium was isolated and cultured from petroleum reservoir in northeast China. Isolate was screened for biosurfactant production using haemolytic assay, Cetyl Trimethyl Ammonium Bromide agar plate assay (CTAB) and the qualitative oil-displacement test. Based on partial sequenced 16S rDNA analysis of isolate, USTBa, identified as Bacillus methylotrophicus with 100% identity. This bacterium was able to produce a type of biosurfactant with excessive foam-forming properties. The maximum biosurfactant production was obtained when the cells were grown on minimal salt medium containing 2% (v/v) crude-oil as the sole source of carbon at 35 °C and 180 rpm after 192 h. This strain had a high emulsification activity and biosurfactant production of 78% and 1.8 g/L respectively. The cell free broth containing biosurfactant could reduce the surface tension to 28 mN/m. Fourier transform infrared (FT-IR) spectrum of extracted biosurfactant indicates the presence of carboxyl, hydroxyl and methoxyl functional groups. Elemental analysis of the biosurfactant by Energy dispersive X-ray spectroscopy (EDS) reveals that the biosurfactant was anionic in nature. The strain USTBa represented as a potent biosurfactant-producer and could be useful in variety of biotechnological and industrial processes, particularly oil industry.     

Characterization and optimization of biosurfactants produced by Acinetobacter baylyi ZJ2 isolated from crude oil-contaminated soil sample toward microbial enhanced oil recovery applications

The present work aims to investigate the surface activity of the biosurfactant produced by Acinetobacter baylyi ZJ2 isolated from crude oil-contaminated soil sample in China and evaluate its potential application in microbial enhanced oil recovery. The biosurfactant produced by A. baylyi ZJ2 was identified as lipopeptide based on thin-layer chromatography, Fourier transform infrared spectroscopy and nuclear magnetic resonance techniques. This biosurfactant could reduce the surface tension of water from 65 mN/m to 35 mN/m, and interfacial tension against oil from 45 mN/m to 15 mN/m. Moreover, surface activity stability results showed that this biosurfactant was effective when the salinity was lower than 8% and the pH value was 4–9, and it was especially stable when the salinity was lower than 4% and pH was 6–7. Based on the result of gas chromatography, there was a decrease in heavy components and an increase in light components, which indicated that A. baylyi ZJ2 exhibited the biodegradability on the heavy components of crude oil. Furthermore, the ability of recovering oil from oil-saturated core showed that nearly 28% additional residual oil was displaced after water flooding. The lipopeptide biosurfactant produced by A. baylyi ZJ2 presented a great potential application in microbial enhanced oil recovery process, owing its good surface activity and satisfying degradation ability to crude oil.     

Biosurfactant production from Candida lipolytica in bioreactor and evaluation of its toxicity for application as a bioremediation agent

This large-scale production, toxicity, characterization and economic analysis of the biosurfactant from Candida lipolytica UCP 0988 produced in the low-medium formulated with animal fat and corn steep liquor was investigated. The biosurfactant was produced in the stationary phase under 200 rpm in the absence of aeration and reduced the surface tension of the medium from 50 to 28 mN/m after 96 h, yielding 10.0 g/L of isolated biosurfactant in a 2 L bioreactor. The production was maximized in a 50 L bioreactor, reaching 40 g/L biosurfactant and 25 mN/m. The cell biomass was quantified and characterized for use in animal nutrition. Chemical structures of the biosurfactant were identified using FTIR and NMR. The crude biosurfactant was not toxic to the bivalve Anomalocardia brasiliana, to the microcrustacean Artemia salina, or three species of vegetables seeds. The biosurfactant stimulated the degradation of motor oil by the seawater indigenous microorganisms. The results obtained indicate that the biosurfactant produced has great potential to be applied as a bioremediation agent for cleaning oil spills.     

Isolation and characterization of biosurfactant production under extreme environmental conditions by alkali-halo-thermophilic bacteria from Saudi Arabia

Twenty three morphologically distinct microbial colonies were isolated from soil and sea water samples, which were collected from Jeddah region, Saudi Arabia for screening of the most potent biosurfactant strains. The isolated bacteria were selected by using different methods as drop collapse test, oil displacement test, blue agar test, blood hemolysis test, emulsification activity and surface tension. The results showed that the ability of Virgibacillus salarius to grow and reduce surface tension under a wide range of pH, salinities and temperatures gives bacteria isolate an advantage in many applications such as pharmaceutical, cosmetics, food industries and bioremediation in marine environment. The biosurfactant production by V. salarius decreased surface tension and emulsifying activity (30 mN/m and 80%, respectively). In addition to reducing the production cost of biosurfactants by tested several plant-derived oils such as jatropha oil, castor oils, jojoba oil, canola oil and cottonseed oil. In this respect the feasibility to reusing old frying oil of sunflower for production rhamnolipids and sophorolipids, their use that lead to solve many ecological and industrial problems.     

Characteristics of biosurfactant produced by Pseudomonas aeruginosa S6 isolated from oil-containing wastewater

A biosurfactant-producing strain S6 was isolated from oil-containing wastewater and identified as Pseudomonas aeruginosa based on physiological and biochemical tests together with 16S rDNA sequence analysis. Thin layer chromatography (TLC) and high-performance liquid chromatography electrospray ionization mass spectra (HPLC-ESI-MS) worked together to reveal that the strain S6 produced rhamnolipid biosurfactant. Mass spectrometry confirmed the presence of some major components in the rhamnolipid surfactant showing m/z of 675.8, 529.6, 503.3 and 475.4, which corresponded to RhaRhaC₁₀C_12:1, RhaC_12:1C₁₀, RhaC₁₀C₁₀ and RhaC₈C₁₀, respectively. The biosurfactant produced by strain S6 had the ability to decrease the surface tension of water from 72 to 33.9 mN m^?1, with the critical micelle concentration (CMC) of 50 mg L^?1. Emulsification experiment indicated that this biosurfactant effectively emulsified the crude petroleum and the measurements of surface tension demonstrated that the biosurfactant possessed stable surface activity at variable ranges of pH and salinity. The biosurfactant also exhibited good performance of phenanthrene solubilization with about 23 times higher solubility of phenanthrene in water than the control. Thus, this biosurfactant may have a potential for application in bioremediation of crude oil contamination.     

Optimization and immobilization of amylase obtained from halotolerant bacteria isolated from solar salterns

The objective of the present study was to isolate halotolerant bacteria from the sediment sample collected from Marakanam Solar Salterns, Tamil Nadu, India using NaCl supplemented media and screened for amylase production. Among the 22 isolates recovered, two strains that had immense potential were selected for amylase production and designated as P1 and P2. The phylogenetic analysis revealed that P1 and P2 have highest homology with Pontibacillus chungwhensis (99%) and Bacillus barbaricus (100%). Their amylase activity was optimized to obtain high yield under various temperature, pH and NaCl concentration. P1 and P2 strain showed respective, amylase activity maximum at 35 °C and 40 °C; pH 7.0 and 8.0; 1.5 M and 1.0 M NaCl concentration. Further under optimized conditions, the amylase activity of P1 strain (49.6 U mL^?1) was higher than P2 strain. Therefore, the amylase enzyme isolated from P. chungwhensis P1 was immobilized in sodium alginate beads. Compared to the free enzyme form (49.6 U mL^?1), the immobilized enzyme showed higher amylase activity as 90.3 U mL^?1. The enzyme was further purified partially and the molecular mass was determined as 40 kDa by SDS–PAGE. Thus, high activity of amylase even under increased NaCl concentration would render immense benefits in food processing industries.     

Variability of soil carbon sequestration capability and microbial activity of different types of salt marsh soils at Chongming Dongtan

Variations in the soil carbon sequestration capability of different types of salt marsh soils at Chongming Dongtan and its influencing factors were studied by analyzing the soil organic carbon (SOC) content, organic matter input and microbial activities. The results indicated that the total SOC content at Area A (southeast of Dongtan, sandy soil with Phragmites communis) was only 46.11% of that of Area B (northeast of Dongtan, clay soil with mixed P. communis and Spartina alterniflora) (P = 0.000 < 0.05), but their organic matter input per year was almost identical. These findings implied that Area B had a lower output of SOC. The microbial biomass at Area A was 3.83 times greater than that at Area B (P = 0.049 < 0.05); the soil catalase and invertase activities at Area A, which were related to carbon metabolism, were 60.31% (P = 0.006 < 0.05) and 34.33% (P = 0.021 < 0.05) higher than at Area B, respectively; and the soil respiration at Area A was also higher than at Area B. These findings implied that the microbial activities at Area A were greater than those at Area B, and therefore the carbon metabolism was rapid, resulting in increased SOC output at Area A. Increased water content and salinity in the clay soil at Area B may inhibit the microbial activities, thereby reducing the decomposition of the organic matter and enhancing carbon sequestration. In addition, some artificial measures for controlling spread of S. alterniflora at Area B (mowing/digging and tillage (M + D); mowing/digging and tillage/waterlogging (M + D + W)) were found to generally improve the microbial activity of soil, thereby increasing SOC output. However, when the two different physical controlling modes were compared, the SOC and microbial activities of the soil subjected to the M + D + W treatment were relatively high and low, respectively, due to waterlogging restraining the microbial metabolism. These findings indicated that the difference in microbial activities was the important factor leading to variability in the SOC sequestration capability between Areas A and B. Additionally, with the exception of soil texture and vegetation types, environmental conditions and artificial turbulence also influenced microbial activities of soil, and hence SOC output and organic carbon sequestration capability.     

Evaluation of fluorescent pseudomonads for plant growth promotion,antifungal activity against Rhizoctonia solani on common bean,and biocontrol potential

Fluorescent pseudomonads are ubiquitous bacteria that are common inhabitants of the rhizosphere and are the most studied group within the genus Pseudomonas. Bacterial isolates (n = 103) from the rhizosphere of wheat and common bean were assessed as potential biocontrol agents in this study. Fungal inhibition tests were performed by a plate assay in which each isolate was tested directly for the production of hydrogen cyanide, protease, siderophore and cellulase. Production of DAPG was verified by using an analytical high performance liquid chromatography assay (HPLC). Plant growth promotion was assessed in phytochamber trials and biocontrol activity was evaluated in greenhouse trials. In all, 52 bacterial isolates with antifungal activity against Rhizoctonia solani were found. Of the 52 isolates, 41 were selected according to their high efficiency in in vitro antagonism, which was shown as inhibition zones in the dual-culture assay. Six of the 41 rhizobacteria, including isolates UTPF7, UTPF13, UTPF18, UTPF22, UTPF27 and strain CHA0 produced HCN. Production of protease enzyme was detected for all isolates excluding UTPF30 isolate. Although some stains appeared not to produce any compound with affinity for ferric iron, other isolates produced prolific amounts, creating a large zone of orange (up to 160 mm², i.e., UTPF16). Seventeen of 41 isolates of fluorescent pseudomonads including strain CHAO produced different amounts of DAPG ranging from 0.6 to 11.4 ng/10⁸ cfu. A total of 39 isolates induced statistically significant effects on plant growth compared with the non-treated control for at least one parameter. The predominant influence observed was increased root length. No bacteria could completely protect the plant against R. solani, although all isolates significantly increased fresh weight as compared to the infested control in greenhouse trials. Pseudomonas fluorescens isolates UTPF16 and UTPF26 significantly (P < 0.05) decreased the number of seedlings with damping-off symptoms in the means of the experiments.     

Isolation of novel chitinolytic bacteria and production optimization of extracellular chitinase

Chitin is one of the most abundant biopolymers widely distributed in the marine and terrestrial environments. Chitinase enzyme has received increased attention due to its wide range of biotechnological applications, especially in agriculture for biocontrol of phytopathogenic fungi and harmful insects. In the present study, 58 bacterial isolates were screened for chitinolytic activity and on the basis of chitin hydrolysis zone 6 isolates were selected for chitinase production in broth media. Based on enzyme production, two most potent isolates identified as Aeromonas hydrophila HS4 and Aeromonas punctata HS6 were selected for further study. The effects of media composition and various fermentation conditions for optimization of chitinase production were studied. The maximum chitinase production was obtained at 37 °C and pH 8.0 after 24–48 h of incubation by HS4; and at 37 °C and pH 7 after 48 h incubation by HS6. Among the substrates colloidal chitin was the best for both the strains. Regarding carbon sources, starch (1%) was the best for both strains; while malt and yeast extract (1%) was found as the best nitrogen source for HS4 and HS6, respectively. Out of metal ions Mn²⁺ and Cu²⁺ enhanced enzyme production in the case of HS6. However, Co²⁺ was the most appropriate for HS4.     

Determination of the genetic similarities of fingerprints from Escherichia coli O157:H7 isolated from different sources in the North West Province,South Africa using ISR,BOXAIR and REP-PCR analysis

The aim of this study was to determine the genetic relationships of Escherichia coli O157:H7 isolated from pigs, cattle, pork, beef, humans and water samples using REP, ISR and BOXAIR PCR analysis. A total of 94 isolates were subjected to the REP-PCR analysis while 95 were screened for ISR and BOXAIR PCR fingerprints. The band sizes for amplicons from the ISR-PCR analysis ranged from 0.173 kb to 0.878 kb. However, a large proportion of the isolates had four bands ranging from 0.447 kb to 0.878 kb. Cluster analysis of the BOXAIR PCR profiles based on banding patterns revealed seven main clusters. It was identified in the clusters III, IV and VII in the BOXAIR PCR that 17.9%, 16.8% and 18.9%, of E. coli O157:H7 isolates respectively were present from all the animal species, meat and water samples. REP-PCR analysis produced 9 different patterns with bands ranging from 0 to 12 per isolate. The band sizes ranged from 200 bp to 8000 bp. Nine major clusters (I–IX) were identified. From the three different species sampled cluster eight was the largest and a mixed cluster with 23.4% (22/94) of the E. coli O157:H7 isolates. These indicate that food products obtained from supermarkets in the study area are contaminated with E. coli O157:H7.     

Purification and characterization of cellulase from a marine Bacillus sp. H1666: A potential agent for single step saccharification of seaweed biomass

In this study, 115 marine bacterial isolates were screened for cellulase enzymatic activity and enzyme with a molecular mass of 40 kDa was purified from culture supernatant of the marine bacterium Bacillus sp. H1666 using ion exchange and size exclusion chromatography method. Growth of bacterial strain H1666 with efficient cellulase enzyme production was observed on untreated wheat straw and rice bran. The biochemical properties of the extracted cellulase were studied and enzyme was found active over a range of pH 3–9. The optimum cellulase activity was observed at pH 7 and temperature 50 °C. The enzyme was also shown to be slightly thermo-stable with 40% residual activity at 60 °C for 4 h. The potential applicability of enzyme was tested on dried green seaweed (Ulva lactuca) and 450 mg/g increase in glucose yield was obtained after saccharification. MALDI TOF–TOF analysis of cellulase peptide fingerprint showed similarity to the sequence of the glycoside hydrolase family protein.     

Effects of grassland conversion to cropland and forest on soil organic carbon and dissolved organic carbon in the farming-pastoral ecotone of Inner Mongolia

Effects of grassland conversion to cropland and forest on soil organic carbon (SOC), dissolved organic carbon (DOC) in the farming-pastoral ecotone of Inner Mongolia were investigated by direct field sampling. SOC content and DOC content in soil decreased after grassland were shifted to forest or cropland, in the sequence of grassland soil > forest soil > cropland soil. SOC stock declined by 18% after grassland shifted from to forest. Reclamation of cropland for 10 years, 15 years and 20 years lost SOC in 0–30 cm soil layer, by 34%, 14% and 18%, respectively, compared with that of grassland. DOC in 3 soil layers was within 21.1–26.5 mg/L in grassland, 12.1–14.6 mg/L in forest soil, and 8.0–14.0 mg/L in cropland soil. Correlation analysis indicated that SOC content and DOC content were positively dependent on total nitrogen content (p < 0.05), but negatively on bulk density or land use type (p < 0.05). DOC was positively correlated SOC (p < 0.01). Moreover, SOC content could be quantitatively described by a linear combination of land use types (p = 0.000, r² = 0.712), and DOC content by a linear combination of two soil-related variables, land use types and SOC (p = 0.000, r² = 0.861).     

On the control of HAB species using low biosurfactant concentrations

Biosurfactants have been suggested as a method to control harmful algal blooms (HABs), but warrant further and more in-depth investigation. Here we have investigated the algicidal effect of a biosurfactant produced by the bacterium Pseudomonas aeruginosa on five diverse marine and freshwater HAB species that have not been tested previously. These include Alexandrium minutum (Dinophycaee), Karenia brevis (Dinophyceae), Pseudonitzschia sp. (Bacillariophyceae), in marine ecosystems, and Gonyostomum semen (Raphidophyceae) and Microcystis aeruginosa (Cyanophyecae) in freshwater. We examined not only lethal but also sub-lethal effects of the biosurfactant. In addition, the effect of the biosurfactant on Daphnia was tested. Our conclusions were that very low biosurfactant concentrations (5 μg mL⁻¹) decreased both the photosynthesis efficiency and the cell viability and that higher concentrations (50 μg mL⁻¹) had lethal effects in four of the five HAB species tested. The low concentrations employed in this study and the diversity of HAB genera tested suggest that biosurfactants may be used to either control initial algal blooms without causing negative side effect to the ecosystem, or to provoke lethal effects when necessary.     

Growth and bioactive secondary metabolites of arctic Protoceratium reticulatum (Dinophyceae)

Harmful algal blooms are mainly caused by marine dinoflagellates and are known to produce potent toxins that may affect the ecosystem, human activities and health. Such events have increased in frequency and intensity worldwide in the past decades. Numerous processes involved in Global Change are amplified in the Arctic, but little is known about species specific responses of arctic dinoflagellates. The aim of this work was to perform an exhaustive morphological, phylogenetical and toxinological characterization of Greenland Protoceratium reticulatum and, in addition, to test the effect of temperature on growth and production of bioactive secondary metabolites. Seven clonal isolates, the first isolates of P. reticulatum available from arctic waters, were phylogenetically characterized by analysis of the LSU rDNA. Six isolates were further characterized morphologically and were shown to produce both yessotoxins (YTX) and lytic compounds, representing the first report of allelochemical activity in P. reticulatum. As shown for one of the isolates, growth was strongly affected by temperature with a maximum growth rate at 15 °C, a significant but slow growth at 1 °C, and cell death at 25 °C, suggesting an adaptation of P. reticulatum to temperate waters. Temperature had no major effect on total YTX cell quota or lytic activity but both were affected by the growth phase with a significant increase at stationary phase. A comparison of six isolates at a fixed temperature of 10 °C showed high intraspecific variability for all three physiological parameters tested. Growth rate varied from 0.06 to 0.19 d⁻¹, and total YTX concentration ranged from 0.3 to 15.0 pg  YTX cell⁻¹ and from 0.5 to 31.0 pg YTX cell⁻¹ at exponential and stationary phase, respectively. All six isolates performed lytic activity; however, for two isolates lytic activity was only detectable at higher cell densities in stationary phase.     

The hydrolysate of barley straw containing inhibitors can be used to produce cephalosporin C by solvent extraction using ethyl acetate

When the solvent extraction of the hydrolysate from barley straw was performed using ethyl acetate (EA), the logarithm of the partition coefficient (log P) of the phenols and furans for EA was found to be more than 1.00, which means that more than 90% of the inhibitors were removed from the hydrolysate layer. Cephalosporin C (CPC) was produced from the hydrolysate of dilute acid pretreatment (DAP) by Acremonium chrysogenum M35. A. chrysogenum M35 was cultured using the hydrolysate and the amount of CPC produced was found to be 10.35 g/L at 144 h. Also, the dry cell weight was about 101.5 g/L at 120 h. The utilization of the hydrolysate for CPC production was effective and the solvent extraction method for the removal of inhibitory substances could contribute to the biorefinery process.     

Esculentin-2CHa: A host-defense peptide with differential cytotoxicity against bacteria,erythrocytes and tumor cells

The host-defense peptide, esculentin-2CHa (GFSSIFRGVA¹⁰KFASKGLGK D²⁰LAKLGVDLVA³⁰ CKISKQC) shows potent (MIC ≤ 6 μM) growth inhibitory activity against clinical isolates of multidrug-resistant strains of Staphylococcus aureus, Acinetobacter baumannii, and Stenotrophomonas maltophilia and differential cytotoxic activity against human erythrocytes (LC₅₀ = 150 μM) and human non-small cell lung adenocarcinoma A549 cells (LC₅₀ = 10 μM). Esculentin-2CHa significantly (P < 0.01) stimulates the release of the anti-inflammatory cytokine IL-10 by mouse lymphoid cells and elevates its production after stimulation with concanavalin A and significantly (P < 0.05) stimulates TNF-α production by peritoneal macrophages. Effects on IL-6 and IL-1β production were not significant. Removal of the hydrophobic N-terminal hexapeptide (GFSSIF) from esculentin-2CHa results in abolition of growth inhibitory activity against S. aureus and cytotoxic activity against erythrocytes and A549 cells as well as a marked (≥16-fold) reduction in potency against A. baumannii and S. maltophilia. The primary structure of esculentin-2 has been poorly conserved between frog species but evolutionary pressure has acted to maintain the hydrophobic character of this N-terminal hexapeptide sequence. Removal of the cyclic C-terminal domain (CKISKQC) and replacement of the Cys³¹ and Cys³⁷ residues by serine resulted in appreciable decreases in cytotoxicity against all microorganisms and against mammalian cells. The more cationic [D20K, D27K] analog showed a modest increase in potency against all microorganisms (up to 4-fold) but a marked increase in cytotoxicity against erythrocytes (LC₅₀ = 11 μM) and A549 cells (LC₅₀ = 3 μM).     

Synthesis and antimicrobial activity of novel amphiphilic aromatic amino alcohols

We report in this work the preparation and in vitro antimicrobial evaluation of novel amphiphilic aromatic amino alcohols synthesized by reductive amination of 4-alkyloxybenzaldehyde with 2-amino-2-hydroxymethyl-propane-1,3-diol. The antibacterial activity was determined against four standard strains (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa) and 21 clinical isolates of methicillin-resistant Staphylococcus aureus. The antifungal activity was evaluated against four yeast (Candida albicans, Candida tropicalis, Candida glabrata and Candida parapsilosis). The results obtained showed a strong positive correlation between the lipophilicity and the antibiotic activity of the tested compounds. The best activities were obtained against the Gram-positive bacteria (MIC = 2–16 μg ml^?1) for the five compounds bearing longer alkyl chains (4c–g; 8–14 carbons), which were also the most active against Candida (MIC = 2–64 μg ml^?1). Compound 4e exhibited the highest levels of inhibitory activity (MIC = 2–16 μg ml^?1) against clinical isolates of MRSA. A concentration of twice the MIC resulted in bactericidal activity of 4d against 19 of the 21 clinical isolates.     

Production and partial characterization of biosurfactant produced by crude oil degrading bacteria

Production of biosurfactant by crude oil degrading bacteria for use in microbial enhanced oil recovery was investigated. Crude oil utilizing bacteria were isolated from soil by enrichment method on oil agar at 30 °C for 5 days. The isolates were identified and screened for biosurfactant production using blood haemolysis and emulsification tests. IR and GC–MS analyses were carried out to detect the type of biosurfactant. The biosurfactant was purified and its stability at various pH, temperature and salinity levels was studied. The organisms were identified as: Achromobacter xylosoxidans subspecies xylosoxidans, Bacillus licheniformis, Proteus vulgaris, Proteus mirabilis, Serratia marcescens, Sphingomonas paucimobilis and Micrococcus kristinae. Emulsification test (E₂₄) revealed that Serratia marcescens had the highest emulsification index of 87%. GC–MS indicated the biosurfactants as lipopeptides. The biosurfactant can be used in EOR under various environmental conditions.     

Soil organic carbon fractions are affected by different land uses in an agro-pastoral transitional zone in Northeastern China

Soil organic carbon (SOC) consists of various C fractions with different stabilities and chemical compositions that are differently affected by changes in land use. A better understanding of the responses of different C fractions to land uses is vital for maintaining soil quality and mitigating global warming. Using data from a short-term land use experiment in northeastern China, this paper investigated the effect of five land uses, corn cropland (Corn), alfalfa grassland (Alfalfa), artificial grassland of Lyemus chinensis (AG), Lyemus chinensis grassland for mowing (AG + Mow) and restored grassland (RG), on the dynamics of total SOC and four SOC fractions with increasing degrees of oxidizability at 0 to 50 cm depths. The results show that land use had a significant effect (P < 0.05) on the total SOC and SOC fractions of very labile C (F1), labile C (F2) and less labile C (F3), while the difference in recalcitrant C (F4) was less pronounced. SOC in the study area was characterized by a predominantly very labile C fraction, and the percentages of F1 to total SOC were more than 40% for all land uses. Compared with Corn, the treatments AG + Mow, AG and RG decreased the percentage of F1 to SOC (by 4.49%, 6.53% and 3.55%, respectively) and increased the percentages of F2 (by 3.32%, 2.77% and 6.60%, respectively) and F3 (by 4.47%, 3.46% and 0.3%, respectively) to SOC. These findings suggest that land-use type is a major factor that influences soil C fractions and that labile C fractions contribute a large part of the total SOC. In addition, grassland colonization of croplands improves soil C sequestration in northeastern China.