Transformation mechanism of carbon tetrachloride and the associated micro-ecology in landfill cover, a typical functional layer zone / 生物工程学报

Landfill is one of the important sources of carbon tetrachloride (CT) pollution, and it is important to understand the degradation mechanism of CT in landfill cover for better control. In this study, a simulated landfill cover system was set up, and the biotransformation mechanism of CT and the associated micro-ecology were investigated. The results showed that three stable functional zones along the depth, i.e., aerobic zone (0-15 cm), anoxic zone (15-45 cm) and anaerobic zone (> 45 cm), were generated because of long-term biological oxidation in landfill cover. There were significant differences in redox condition and microbial community structure in each zone, which provided microbial resources and favorable conditions for CT degradation. The results of biodegradation indicated that dechlorination of CT produced chloroform (CF), dichloromethane (DCM) and Cl- in anaerobic and anoxic zones. The highest concentration of dechlorination products occurred at 30 cm, which were degraded rapidly in aerobic zone. In addition, CT degradation rate was 13.2-103.6 μg/(m2·d), which decreased with the increase of landfill gas flux. The analysis of diversity sequencing revealed that Mesorhizobium, Thiobacillus and Intrasporangium were potential CT-degraders in aerobic, anaerobic and anoxic zone, respectively. Moreover, six species of dechlorination bacteria and eighteen species of methanotrophs were also responsible for anaerobic transformation of CT and aerobic degradation of CF and DCM, respectively. Interestingly, anaerobic dechlorination and aerobic transformation occurred simultaneously in the anoxic zone in landfill cover. Furthermore, analysis of degradation mechanism suggested that generation of stable anaerobic-anoxic-aerobic zone by regulation was very important for the harmless removal of full halogenated hydrocarbon in vadose zone, and the increase of anoxic zone scale enhanced their removal. These results provide theoretical guidance for the removal of chlorinated pollutants in landfills.

Advances in microbial degradation of chlorinated hydrocarbons / 生物工程学报

Chlorinated hydrocarbons (CAHs) threaten human health and the ecological environment due to their strong carcinogenic, teratogenic, mutagenic and heritable properties. Heterotrophic assimilation degradation can completely and effectively degrade CAHs, without secondary pollution. However, it is crucial to comprehensively understand the heterotrophic assimilation process of CAHs for its application. Therefore, we review here the characteristics and advantages of heterotrophic assimilation degradation of CAHs. Moreover, we systematically summarize current research status of heterotrophic assimilation of CAHs. Furthermore, we analyze bacterial genera and metabolism, key enzymes and characteristic genes involved in the metabolic process. Finally, we indicate existing problems of heterotrophic assimilation research and future research needs.

Advances in biotic and abiotic mutual promoting mechanism for chlorinated aliphatic hydrocarbons degradation / 生物工程学报

Chlorinated aliphatic hydrocarbons (CAHs) with characteristics of high toxicity, biological accumulation and recalcitrance to degradation as well as carcinogenicity, teratogenesis and mutagenicity, are seriously harmful to human health and ecological environment. CAHs degradation depends on biotic and abiotic responses that exist diversified interactive effects, so it is important to clarify the mechanism of CAHs degradation via biotic and abiotic mutual promoting to significantly enhance the CAHs-contaminated site restoration. In this work, a series of pathways for CAHs degradation was first introduced and summarized as three means on reductive dechlorination, aerobic cometabolism and direct oxidation, and biotic and abiotic typical factors affecting CAHs degradation were concluded from these. Then, mechanisms of induced degradation and synergistic degradation were indicated from the perspective of mutual promoting degradation both with biotic and abiotic responses, and furthermore, the application and technical limitations of CAHs degradation enhanced via biotic and abiotic mutual promoting were reviewed and analyzed. Finally, the development of CAHs degradation technology in future was prospected.

Effects of copper on biodegradation mechanism of trichloroethylene by mixed microorganisms / 生物工程学报

We isolated and enriched mixed microorganisms SWA1 from landfill cover soils supplemented with trichloroethylene (TCE). The microbial mixture could degrade TCE effectively under aerobic conditions. Then, we investigated the effect of copper ion (0 to 15 μmol/L) on TCE biodegradation. Results show that the maximum TCE degradation speed was 29.60 nmol/min with 95.75% degradation when copper ion was at 0.03 μmol/L. In addition, genes encoding key enzymes during biodegradation were analyzed by Real-time quantitative reverse transcription PCR (RT-qPCR). The relative expression abundance of pmoA gene (4.22E-03) and mmoX gene (9.30E-06) was the highest when copper ion was at 0.03 μmol/L. Finally, we also used MiSeq pyrosequencing to investigate the diversity of microbial community. Methylocystaceae that can co-metabolic degrade TCE were the dominant microorganisms; other microorganisms with the function of direct oxidation of TCE were also included in SWA1 and the microbial diversity decreased significantly along with increasing of copper ion concentration. Based on the above results, variation of copper ion concentration affected the composition of SWA1 and degradation mechanism of TCE. The degradation mechanism of TCE included co-metabolism degradation of methanotrophs and oxidation metabolism directly at copper ion of 0.03 μmol/L. When copper ion at 5 μmol/L (biodegradation was 84.75%), the degradation mechanism of TCE included direct-degradation and co-metabolism degradation of methanotrophs and microorganisms containing phenol hydroxylase. Therefore, biodegradation of TCE by microorganisms was a complicated process, the degradation mechanism included co-metabolism degradation of methanotrophs and bio-oxidation of non-methanotrophs.

Sequence analysis of 16S rDNA and pmoCAB gene cluster of trichloroethylene-degrading methanotroph / 生物工程学报

Methanotrophs could degrade methane and various chlorinated hydrocarbons. The analysis on methane monooxygenase gene cluster sequence would help to understand its catalytic mechanism and enhance the application in pollutants biodegradation. The methanotrophs was enriched and isolated with methane as the sole carbon source in the nitrate mineral salt medium. Then, five chlorinated hydrocarbons were selected as cometabolic substrates to study the biodegradation. The phylogenetic tree of 16S rDNA using MEGE5.05 software was constructed to identify the methanotroph strain. The pmoCAB gene cluster encoding particulate methane monooxygenase (pMMO) was amplified by semi-nested PCR in segments. ExPASy was performed to analyze theoretical molecular weight of the three pMMO subunits. As a result, a strain of methanotroph was isolated. The phylogenetic analysis indicated that the strain belongs to a species of Methylocystis, and it was named as Methylocystis sp. JTC3. The degradation rate of trichloroethylene (TCE) reached 93.79% when its initial concentration was 15.64 μmol/L after 5 days. We obtained the pmoCAB gene cluster of 3 227 bp including pmoC gene of 771 bp, pmoA gene of 759 bp, pmoB gene of 1 260 bp and two noncoding sequences in the middle by semi-nested PCR, T-A cloning and sequencing. The theoretical molecular weight of their corresponding gamma, beta and alpha subunit were 29.1 kDa, 28.6 kDa and 45.6 kDa respectively analyzed using ExPASy tool. The pmoCAB gene cluster of JTC3 was highly identical with that of Methylocystis sp. strain M analyzed by Blast, and pmoA sequences is more conservative than pmoC and pmoB. Finally, Methylocystis sp. JTC3 could degrade TCE efficiently. And the detailed analysis of pmoCAB from Methylocystis sp. JTC3 laid a solid foundation to further study its active sites features and its selectivity to chlorinated hydrocarbon.

Advances in degradation of chlorinated hydrocarbons by obligate and facultative methanotrophs / 生物工程学报

Bioremediation is one of the most effective ways to treat and dispose of chlorinated hydrocarbons, and methanotrophs are potentially useful to do so. Recent studies found that facultative methanotrophs can use compounds containing C-C bond as sources of carbon and energy, thus overcoming the limitation that obligate methanotrophsone uses only C1 compounds for this process. This is a unique metabolic approach that is becoming increasingly attractive in the field of contaminant biodegradation. Here, we summarized the bioremediation of chlorinated hydrocarbons by obligate and facultative methanotrophs. This process involves the degradation of various chlorinated hydrocarbons by diverse strains, including pure cultures and mixed cultures. We also compare the activity expression and catalytic properties of different types of methane monooxygenases in various substrates. We furthermore summarize the kinetic characteristics of the degradation of chlorinated hydrocarbons using the model strain Methylosinus trichosporium OB3b, and outline the degradation and potential of chlorinated hydrocarbons by facultative methanotrophs. Lastly, we discuss current problems and future research directions for degradation of chlorinated hydrocarbons by methanotrophs.