Yu-Hua Zhao

Zhejiang University, Hang-hsien, Zhejiang Sheng, China

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Publications (28)62.79 Total impact

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    ABSTRACT: Azo dyes are recalcitrant and refractory pollutants that constitute a significant menace to the environment. The present study is focused on exploring the capability of Bacillus sp. strain UN2 for application in methyl red (MR) degradation. Effects of physicochemical parameters (pH of medium, temperature, initial concentration of dye, and composition of the medium) were studied in detail. The suitable pH and temperature range for MR degradation by strain UN2 were respectively 7.0-9.0 and 30-40 °C, and the optimal pH value and temperature were respectively 8.0 and 35 °C. Mg(2+) and Mn(2+) (1 mM) were found to significantly accelerate the MR removal rate, while the enhancement by either Fe(3+) or Fe(2+) was slight. Under the optimal degradation conditions, strain UN2 exhibited greater than 98 % degradation of the toxic azo dye MR (100 ppm) within 30 min. Analysis of samples from decolorized culture flasks confirmed biodegradation of MR into two prime metabolites: N,N'dimethyl-p-phenyle-nediamine and 2-aminobenzoic acid. A study of the enzymes responsible for the biodegradation of MR, in the control and cells obtained during (10 min) and after (30 min) degradation, showed a significant increase in the activities of azoreductase, laccase, and NADH-DCIP reductase. Furthermore, a phytotoxicity analysis demonstrated that the germination inhibition was almost eliminated for both the plants Triticum aestivum and Sorghum bicolor by MR metabolites at 100 mg/L concentration, yet the germination inhibition of parent dye was significant. Consequently, the high efficiency of MR degradation enables this strain to be a potential candidate for bioremediation of wastewater containing MR.
    Environmental Science and Pollution Research 01/2014; · 2.76 Impact Factor
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    ABSTRACT: Azoreductase (AZR) has been found to be an effective enzyme for azo dye removal. However, NAD(P)H is required as electron donor for the cleavage of the azo bond by AZR. Due to the high cost of the reduced nicotinamide coenzyme, the large scale application of AZR is impractical. In this study, an artificial fusion enzyme of azoreductase and alkaline-resistant NAD(P)-dependent glucose 1-dehydrogenase (LsGDH) called GFA was constructed for NAD(P)H regeneration and cyclic utilization. The purified fusion enzyme migrated as a single protein band on SDS–PAGE with a molecular weight of ∼55 kDa, exhibiting both azoreductase and glucose 1-dehydrogenase activities. The azoreductase moiety of the fusion enzyme exhibited high enzyme activities towards Methyl Red (160.62 U mg−1), which was slightly higher than parental azoreductase (153.16 U mg−1). Compared with parental enzymes, the azoreductase moiety was characterized by kinetic parameters of decreased Km (0.52-fold) and increased Kcat (1.25-fold). These indicate a 2.41-fold net increase in catalytic efficiency (Kcat/Km) of the azoreductase moiety. The highest enzymatic degradation was achieved at the temperature of 45 °C and pH of 7.0, respectively. Mg2+ and Ca2+ enhanced the fusion enzyme activity, while Cr2+, Mn2+and Zn2+ inhibited the activity to varying extents. The enzyme activity of the fusion enzyme was enhanced by low concentrations of dodecyl trimethyl ammonium bromide (DTAB) and cetylpyridinium chloride (CPC) but inhibited by high concentrations of surfactants.
    International Biodeterioration & Biodegradation 01/2014; 87:81–86. · 2.06 Impact Factor
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    ABSTRACT: An understanding of the genetic basis underlying the phenotypic variations of yeast strains would guide the breeding of this useful microorganism. Here, comparative functional genomics (CFG) of two bioethanol Saccharomyces cerevisiae strains (YJS329 and ZK2) with different stress tolerances and ethanol fermentation performances were performed. Our analysis indicated that different patterns of gene expression in the central carbon metabolism, antioxidative factors, and membrane compositions of these two strains are the main contributors to their various traits. Some of the differently expressed genes were directly caused by the genomic structural variations between YJS329 and ZK2. Moreover, CFG of these two strains also led to novel insights into the mechanism of stress tolerance in yeast. For example, it was found that more oleic acid in the plasma membrane contributes to the acetic acid tolerance of yeast. Based on the genetic information particular to each strain, strategies to improve their adaptability and ethanol fermentation performances were designed and confirmed. Thus, CFG could not only help reveal basis of phenotypic diversities but also guide the genetic breeding of industrial microorganisms.
    Applied Microbiology and Biotechnology 01/2013; · 3.69 Impact Factor
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    ABSTRACT: Various types of lignocellulosic wastes extensively used in biofuel production were provided to assess the potential of EXLX1 as a cellulase synergist. Enzymatic hydrolysis of natural wheat straw showed that all the treatments using mixtures of cellulase and an optimized amount of EXLX1, released greater quantities of sugars than those using cellulase alone, regardless of cellulase dosage and incubation time. EXLX1 exhibited different synergism and binding characteristics for different wastes, but this can be related to their lignocellulosic components. The cellulose proportion could be one of the important factors. However, when the cellulose proportion of different biomass samples exhibited no remarkable differences, a higher synergism of EXLX1 is prone to occur on these materials, with a high proportion of hemicellulose and a low proportion of lignin. The information could be favorable to assess whether EXLX1 is effective as a cellulase synergist for the hydrolysis of the used materials. Binding assay experiments further suggested that EXLX1 bound preferentially to alkali pretreated materials, as opposed to acid pretreated materials under the assay condition and the binding preference would be affected by incubation temperature.
    PLoS ONE 01/2013; 8(9):e75022. · 3.73 Impact Factor
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    ABSTRACT: BACKGROUND: Environmental stresses and inhibitors encountered by Saccharomyces cerevisiae strains are the main limiting factors in bioethanol fermentation. Strains with different genetic backgrounds usually show diverse stress tolerance responses. An understanding of the mechanisms underlying these phenotypic diversities within S. cerevisiae populations could guide the construction of strains with desired traits. RESULTS: We explored the genetic characteristics of the bioethanol S. cerevisiae strain YJS329 and elucidated how genetic variations in its genome were correlated with specified traits compared to similar traits in the S288c-derived strain, BYZ1. Karyotypic electrophoresis combined with array-comparative genomic hybridization indicated that YJS329 was a diploid strain with a relatively constant genome as a result of the fewer Ty elements and lack of structural polymorphisms between homologous chromosomes that it contained. By comparing the sequence with the S288c genome, a total of 64,998 SNPs, 7,093 indels and 11 unique genes were identified in the genome of YJS329-derived haploid strain YJSH1 through whole-genome sequencing. Transcription comparison using RNA-Seq identified which of the differentially expressed genes were the main contributors to the phenotypic differences between YJS329 and BYZ1. By combining the results obtained from the genome sequences and the transcriptions, we predicted how the SNPs, indels and chromosomal copy number variations may affect the mRNA expression profiles and phenotypes of the yeast strains. Furthermore, some genetic breeding strategies to improve the adaptabilities of YJS329 were designed and experimentally verified. CONCLUSIONS: Through comparative functional genomic analysis, we have provided some insights into the mechanisms underlying the specific traits of the bioenthanol strain YJS329. The work reported here has not only enriched the available genetic resources of yeast but has also indicated how functional genomic studies can be used to improve genetic breeding in yeast.
    BMC Genomics 09/2012; 13(1):479. · 4.40 Impact Factor
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    ABSTRACT: Bensulfuron-methyl (BSM) is a new kind of sulfonylurea herbicide widely used to control broad-leaf weeds in rice paddies. The aim of this work was to study BSM biodegradation in paddy soils with BSM-degrading bacteria Bacillus megaterium L1 and Brevibacterium sp. BH and its effect on the structures of soil bacterial community. More than 90 % of BSM could be degraded in paddy soils with 0.0355 mg kg⁻¹ BSM concentration. Addition of BSM-degrading bacterial strains Bacillus megaterium L1 into BSM contaminated paddy soil could have the half-life time of BSM compared to treatment without Bacillus megaterium L1 inoculation. Denaturing gradient gel electrophoresis and principle component analysis indicated that the diversity of the soil microbial community structure changed along with the addition of BSM, which recovered at the end of the experiment (5 weeks). Addition of BSM-degrading bacteria Bacillus megaterium L1 enriched the diversity of soil microbial community structure in paddy soils. This study provides information on the biodegradation of BSM and BSM's influences on the soil bacteria microbial community structures.
    Ecotoxicology 03/2012; 21(5):1281-90. · 2.77 Impact Factor
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    ABSTRACT: A phosphite dehydrogenase gene (ptdhK) consisting of 1,011-bp nucleotides which encoding a peptide of 336 amino acid residues was cloned from Pseudomonas sp. K. gene ptdhK was expressed in Escherichia coli BL21 (DE3) and the corresponding recombinant enzyme was purified by metal affinity chromatography. The recombinant protein is a homodimer with a monomeric molecular mass of 37.2 kDa. The specific activity of PTDH-K was 3.49 U mg(-1) at 25 °C. The recombinant PTDH-K exhibited maximum activity at pH 3.0 and at 40 °C and displayed high stability within a wide range of pHs (5.0 to 10.5). PTDH-K had a high affinity to its natural substrates, with K (m) values for sodium phosphite and NAD of 0.475 ± 0.073 and 0.022 ± 0.007 mM, respectively. The activity of PTDH-K was enhanced by Na(+), NH (4) (+) , Mg(2+), Fe(2+), Fe(3+), Co(2+), and EDTA, and PTDH-K exhibited different tolerance to various organic solvents.
    Applied biochemistry and biotechnology 03/2012; 166(5):1301-13. · 1.94 Impact Factor
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    ABSTRACT: The ability of Pseudomonas sp. strain DY1 to adsorb Acid Black 172 was studied to determine the kinetics and mechanisms involved in biosorption of the dye. Kinetic data for adsorption fit a pseudo-second-order model. Increased initial dye concentration could significantly enhance the amount of dye adsorbed by heat-treated biomass in which the maximum amount of dye adsorbed was as high as 2.98 mmol/g biomass, whereas it had no significant influence on dye sorption by live biomass. As treated temperature increased, the biomass showed gradual increase of dye sorption ability. Experiments using potentiometric titration and Fourier transform infrared spectroscopy (FTIR) indicated that amine groups (NH2) played a prominent role in biosorption of Acid Black 172. Scanning electron microscopy (SEM), atomic force microscopy (AFM) and transmission electron microscopy (TEM) analysis indicated that heat treatment of the biomass increased the permeability of the cell walls and denatured the intracellular proteins. The results of biosorption experiments by different cell components confirmed that intracellular proteins contributed to the increased biosorption of Acid Black 172 by heat-treated biomass. The data suggest that biomass produced by this strain may have application for removal of metal-complex dyes from wastewater streams generated from the dye products industry.
    Journal of hazardous materials 02/2012; 205-206:47-54. · 4.14 Impact Factor
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    ABSTRACT: Malachite Green (MG) is used for a variety of applications but is also known to be carcinogenic and mutagenic. In this study, a novel Micrococcus sp. (strain BD15) was observed to efficiently decolorize MG. The purposes of this study were to explore the optimal conditions for decolorization and to evaluate the potential use of this strain for MG decolorization. Optical microscope and UV-visible analyses were carried out to determine whether the decolorization was due to biosorption or biodegradation. A Plackett-Burman design was employed to investigate the effect of various parameters on decolorization, and response surface methodology was then used to explore the optimal decolorization conditions. Kinetics analysis and antimicrobial activity tests were also performed. The results indicated that the decolorization by the strain was mainly due to biodegradation. Concentrations of MG, urea, and yeast extract and inoculum size had significantly positive effects on MG decolorization, while concentrations of CuCl(2) and MgCl(2), and temperature had significantly negative effects. The interaction between different parameters could significantly affect decolorization, and the optimal conditions for decolorization were 1.0 g/L urea, 0.9 g/L yeast extract, 100 mg/L MG, 0.1 g/L inoculums (dry weight), and incubation at 25.2°C. Under the optimal conditions, 96.9% of MG was removed by the strain within 1 h, which represents highly efficient microbial decolorization. Moreover, the kinetic data for decolorization fit a second-order model well, and the strain showed a good MG detoxification capability. Based on the results of this study, we propose Micrococcus sp. strain BD15 as an excellent candidate strain for MG removal from wastewater.
    Environmental Science and Pollution Research 02/2012; 19(7):2898-907. · 2.76 Impact Factor
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    ABSTRACT: The performances of nine biosorbents derived from dead fungal biomass were investigated for their ability to remove Reactive Black 5 from aqueous solution. The biosorption data for removal of Reactive Black 5 were readily modeled using the Langmuir adsorption isotherm. Kinetic analysis based on both pseudo-second-order and Weber-Morris models indicated intraparticle diffusion was the rate limiting step for biosorption of Reactive Black 5 on to the biosorbents. Sorption capacities of the biosorbents were not correlated with the initial biosorption rates. Sensitivity analysis of the factors affecting biosorption examined by an artificial neural network model showed that pH was the most important parameter, explaining 22%, followed by nitrogen content of biosorbents (16%), initial dye concentration (15%) and carbon content of biosorbents (10%). The biosorption capacities were not proportional to surface areas of the sorbents, but were instead influenced by their chemical element composition. The main functional groups contributing to dye sorption were amine, carboxylic, and alcohol moieties. The data further suggest that differences in carbon and nitrogen contents of biosorbents may be used as a selection index for identifying effective biosorbents from dead fungal biomass.
    PLoS ONE 01/2012; 7(3):e33551. · 3.73 Impact Factor
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    ABSTRACT: Two lipopeptide antibiotics, pelgipeptins C and D, were isolated from Paenibacillus elgii B69 strain. The molecular masses of the two compounds were both determined to be 1,086 Da. Mass-spectrometry, amino acid analysis and NMR spectroscopy indicated that pelgipeptin C was the same compound as BMY-28160, while pelgipeptin D was identified as a new antibiotic of the polypeptin family. These two peptides were active against all the tested microorganisms, including antibiotic-resistant pathogenic bacterial strains such as methicillin-resistant Staphylococcus aureus (MRSA). Time-kill assays demonstrated that pelgipeptin D exhibited rapid and effective bactericidal action against MRSA at 4×MIC. Based on acute toxicity test, the intraperitoneal LD50 value of pelgipeptin D was slightly higher than that of the structurally related antimicrobial agent polymyxin B. Pelgipeptins are highly potent antibacterial and antifungal agents, particularly against MRSA, and warrant further investigation as possible therapeutic agents for bacteria infections resistant to currently available antibiotics.
    The Journal of Microbiology 12/2011; 49(6):942-9. · 1.28 Impact Factor
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    ABSTRACT: Malachite green (MG), a widely-used and recalcitrant dye, has been confirmed to be carcinogenic and mutagenic against many organisms. The main objective of this study is to investigate the capability of Pseudomonas sp. strain DY1 to decolorize MG, and to explore the possible mechanism. The results showed that this strain demonstrated high decolorizing capability (90.3-97.2%) at high concentrations of MG (100-1,000 mg/l) under shaking condition within 24 h. In static conditions, lower but still effective decolorization (78.9-84.3%) was achieved. The optimal pH and temperature for the decolorization was pH 6.6 and 28-30°C, respectively. Mg(2+) and Mn(2+) (1 mM) were observed to significantly enhance the decolorization. The intermediates of the MG degradation under aerobic condition identified by UV-visible, GC-MS and LC-MS analysis included malachite green carbinol, (dimethyl amino-phenyl)-phenyl-methanone, N,N-dimethylaniline, (methyl amino-phenyl)-phenyl-methanone, (amino phenyl)-phenyl methanone and di-benzyl methane. The enzyme analysis indicated that Mn-peroxidase, NADH-DCIP and MG reductase were involved in the biodegradation of MG. Moreover, phytotoxicity of MG and detoxification for MG by the strain were observed. Therefore, this strain could be potentially used for bioremediation of MG.
    Ecotoxicology 03/2011; 20(2):438-46. · 2.77 Impact Factor
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    ABSTRACT: Acetic acid existing in a culture medium is one of the most limiting constraints in yeast growth and viability during ethanol fermentation. To improve acetic acid tolerance in Saccharomyces cerevisiae strains, a drug resistance marker-aided genome shuffling approach with higher screen efficiency of shuffled mutants was developed in this work. Through two rounds of genome shuffling of ultraviolet mutants derived from the original strain 308, we obtained a shuffled strain YZ2, which shows significantly faster growth and higher cell viability under acetic acid stress. Ethanol production of YZ2 (within 60 h) was 21.6% higher than that of 308 when 0.5% (v/v) acetic acid was added to fermentation medium. Membrane integrity, higher in vivo activity of the H+-ATPase, and lower oxidative damage after acetic acid treatment are the possible reasons for the acetic acid-tolerance phenotype of YZ2. These results indicated that this novel genome shuffling approach is powerful to rapidly improve the complex traits of industrial yeast strains.
    Journal of Industrial Microbiology 03/2011; 38(3):415-22. · 1.80 Impact Factor
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    ABSTRACT: An efficient nitrobenzene degrading bacterium strain NB5, which was able to utilize nitrobenzene as a sole source of carbon, nitrogen and energy under aerobic condition, was isolated from activated sludge in an oil refinery at Hangzhou, China. Based on phenotypic features, 16S rDNA gene sequencing and G + C content analysis, strain NB5 was identified as Rhodococcus sp. NB5. Nitrobenzene degradation experiments using high performance liquid chromatograph (HPLC) showed that strain NB5 could tolerate a high nitrobenzene concentration and completely degrade nitrobenzene with initial concentration ranging from 100 mg · l–1 to 1000 mg · l–1 within 144 h. The optimal degradation and cell growth were observed at 30 °C, pH 7.0. The addition of second nitrogen source (0.1%) such as urea, peptone, yeast extract and beef extract generally enhanced degradation of nitrobenzene. Rhodococcus sp. strain NB5 could be an excellent candidate for biotreatment of industrial wastewater containing high concentration of nitrobenzene. (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
    Journal of Basic Microbiology 02/2011; 51(4):397 - 403. · 1.20 Impact Factor
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    ABSTRACT: The gene gdh encoding an organic solvent-tolerant and alkaline-resistant NAD(P)-dependent glucose 1-dehydrogenase (LsGDH) was cloned from Lysinibacillus sphaericus G10 and expressed in Escherichia coli. The recombinant LsGDH exhibited maximum activity at pH 9.5 and 50 °C. LsGDH displayed high stability at a wide pH ranging from 6.5 to 10.0 and was stable after incubation at 30 °C for 1 week in 25 mM sodium phosphate buffer (pH 6.5) in the absence or presence of NaCl. The activity of LsGDH was enhanced by Li+, Na+, K+, NH4+, Mg2+, and EDTA at pH 8.0. LsGDH exhibited high tolerance to 60% DMSO, 30% acetone, 30% methanol, 30% ethanol, 10% n-propanol, 30% isopropanol, 60% n-hexanol and 30% n-hexane. The relationship between stability and chain length of the alcohols fit a Gaussian distribution model (R2≥0.94), and demonstrated lowest enzyme stability in C4-alcohol. The results suggested that LsGDH was potentially useful for coenzyme regeneration in organic solvents or under alkaline conditions.
    Bioresource Technology 01/2011; 102(2):1528-36. · 5.04 Impact Factor
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    ABSTRACT: Pseudomonas sp. strain DY1 was newly isolated from soil with rotten wood and identified as a member of the genus Pseudomonas based on 16S rDNA and biochemical tests. Acid Black 172, a water soluble Cr-complex dye, was then selected as a model dye to investigate the decolorisation ability of the strain. It was observed that the growth of the strain was not inhibited by high dose of metal ions (10 mM), and efficient decolorisation was still observed when high concentrations of Fe(2+), Fe(3+) and Ca(2+) existed. The optimal decolorising conditions obtained from Taguchi design were as follows: temperature 37˚C, pH 7.0, Fe(3+) and proline concentrations of 7 mM and 3.0 g/L, respectively. Under the optimal conditions, 94.5% of Acid Black 172 (100 mg/L) could be decolorised by the strain in 24 h. The kinetics of the decolorisation best fitted the first order kinetic model (R(2)=0.981). Besides, the phytotoxicity study demonstrated a good detoxification by the strain. This work has a certain practical value in microbial decolorisation of textile wastewater.
    Water Science & Technology 01/2011; 63(7):1531-8. · 1.10 Impact Factor
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    ABSTRACT: Direct microbial conversion of wheat straw into lipid by a cellulolytic fungus of Aspergillus oryzae A-4 in solid-state fermentation (SSF) was investigated. In submerged fermentation, A. oryzae A-4 accumulated lipid to 15-18.15% of biomass when pure cellulose was utilized as the sole substrate. In SSF of the wheat straw and bran mixture, A. oryzae A-4 yielded lipid of 36.6mg/g dry substrate (gds), and a cellulase activity of 1.82 FPU/gds with 25.25% of holocellulose utilization in the substrates were detected on the 6th day. The lipid yield reached 62.87 mg/gds in SSF on the 6th day under the optimized conditions from Plackett-Burman design (PBD). Cellulase secretion of A. oryzae A-4 was found to influence the lipid yield. Dilute acid pretreatment of the straw and addition of some agro-industrial wastes to the straw could enhance lipid production of A. oryzae A-4.
    Bioresource Technology 10/2010; 101(19):7556-62. · 5.04 Impact Factor
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    ABSTRACT: In this study, a systemic analysis was initially performed to investigate the relationship between fermentation-related stress tolerances and ethanol yield. Based on the results obtained, two elite Saccharomyces cerevisiae strains, Z8 and Z15, with variant phenotypes were chosen to construct strains with improved multi-stress tolerance by genome shuffling in combination with optimized initial selection. After three rounds of genome shuffling, a shuffled strain, YZ1, which surpasses its parent strains in osmotic, heat, and acid tolerances, was obtained. Ethanol yields of YZ1 were 3.11%, 10.31%, and 10.55% higher than those of its parent strains under regular, increased heat, and high gravity fermentation conditions, respectively. YZ1 was applied to bioethanol production at an industrial scale. Results demonstrated that the variant phenotypes from available yeast strains could be used as parent stock for yeast breeding and that the genome shuffling approach is sufficiently powerful in combining suitable phenotypes in a single strain.
    Bioresource Technology 10/2010; 102(3):3020-7. · 5.04 Impact Factor
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    ABSTRACT: Based on the potential decolorization ability of Pseudomonas sp. strain DY1, a Plackett–Burman design was used to select the operational parameters for decolorization of heavy metal-containing dye Acid Black 172. Four significant parameters, including pH, temperature, concentrations of Fe3+ and NaH2PO4, were optimized. A quadratic model obtained from the response surface design was constructed on experimental data. The optimal condition for decolorization was found to be pH 6.23, 30 °C, 8.0 mM of Fe3+, and 10.0 g/L of NaH2PO4. The confirmatory experiments (86.0% decolorization percentage within the confidence interval) subsequently verified the accuracy of the experimental model. Moreover, the decolorization under the optimal condition fitted the logarithmic model well (R2 = 0.964). This strain really offers an alternative for bioremediation of soil or water contaminated by heavy metal-containing dye.
    International Biodeterioration & Biodegradation. 01/2010;

Publication Stats

122 Citations
62.79 Total Impact Points

Institutions

  • 2010–2014
    • Zhejiang University
      • College of Life Sciences
      Hang-hsien, Zhejiang Sheng, China
    • China National Rice Research Institute
      Hang-hsien, Zhejiang Sheng, China
  • 2013
    • Zhejiang Academy of Agricultural Sciences
      Zhegang, Jiangxi Sheng, China