The research objective of the thesis is to select microorganisms capable of producing laccase, laccaselike from Ba Vi National Forest area, in herbicide / dioxin contaminated soil at Bien Hoa airport. Assessing the ability of decomposing herbicides containing dioxin and reactive dyes by laccase, laccase-like and VSV strains were selected to guide the application in defense activities.
MINISTRY OF EDUCATION AND TRAINING VIET NAM ACADAMY OF SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY SCIENCE AND TECHNOLOGY - PHUNG KHAC HUY CHU REASEARCH ON THE ABILITY OF DECOLORIZATION OF REACTIVE DYES AND DEGRADATION OF HERBICIDES/DIOXIN BY MICROBES PRODUCING LACCASE Major: Environmental Technique Code: 9.52.03.20 SUMMARRY OF ENVIRONMENTAL TECHNIQUE DOCTORAL THESIS Hanoi, 2018 The work was realized in Graduate University of Sicence and Technology – Vietnam Academy of Science and Technology Advisor 1: Assoc Professor, Dr Dang Thi Cam Ha Advisor 2: Examiner 1:……………………… Examiner 2:……………………… Examiner 3:………………………… The thesis will be evaluated by doctoral committee at Graduate University of Science and Technology, Vietnam Academy of Science and Technology on date… month……2018 The thesis is available at: - Library of Graduate University of Science and Technology - National Library of Vietnam INTRODUCTION The necessary of the thesis In recent years, pollution by chemicals including difficultlydecomposed organic substances (POPs) increasingly created by human beings, causing a lot of implications for human health and the environment In such developing countries as Vietnam, the use and discharge of chemical waste in agriculture, medicine, industry and other manufacturing sectors without ability to be controlled has caused serious consequences to the environment and humans In addition, herbicides containing dioxin in existence by the wars after more than 40 years and the nature of dioxins formed during the process of industrial development still continues to cause more serious consequences for the ecological environment and people in Vietnam Vietnam is a tropical country of biodiversity in the top 10 of the world, especially microbial diversity There are many species of microorganisms with differently-synthesized enzymes; two of which are oxidoreductive and peroxidase enzyme having included many high performance applications in the fields of economy and protecting human health Being not only high-activity laccase but also capable of strong activity, just together with liberal oxygen as well as the help of fastening substance or fastening systems (mediator) Then this laccase has been in the greater consideration than any other Therefore, the subject of study and research with the name "Research on the ability of decolorization of reactive dyes and degradation of herbicides/dioxin by microbes producing laccase" has been conducted 2 Research objectives A selection of microbes capable of laccase, laccase-like strains borning from Ba Vi National forests, in the soil of contaminated herbicides/dioxin at the Bien Hoa formed airbase; Ability to degrade the herbicides containing dioxin and decolor type reactive dyes by laccase, laccase-like strains, and microbe strains were chosen to apply in the orientation in defense activities The implementation content Isolation, classification of fungi, ascomycetes that capable of producing high potential laccase-like and laccase-like from the Ba Vi National Forest and dioxin contaminated soil at Bien Hoa Airbase, Dong Nai Province; selection cultured medium highly capable of laccase, laccase-like biosynthesis, representatives from Basicomycetes and Streptomycetes are highly isolated; Research the chemical-physical characteristics, chemicalbiological basis of laccase, laccase-like are purified; Performance evaluation of decolorization types of synthetic dyes, reactive dyes by laccase, laccase-like strain of microbes and laccase biosynthesis, laccase-like; Evaluate the biodegradable herbicides 2.4-D, 2, 4, 5-T pure and have in land pollution in Bien Hoa formed airbase by laccase, single race and mixed fungal strains laccase biosynthesis; Evaluate the performance of biodegradable type 2, 3, 7, 8-TCDD with single fungal strains and strain mixture laccase biosynthesis CHAPTER LITERATURE REVIEW 1.1 Laccase, laccase-like microbial biosynthesis and laccase, laccase-like 1.1.1 General introduction of laccase Laccase was interested in research from many years with the content: Group microorganisms laccase synthesis, molecular structure and mechanism of catalytic activity, the study of gene, protein and chemical-physical characteristics of laccase Laccase is, through the previous ones, shown to be the subject of quite detailed research There has, so far, been little use of many microbial strains of laccase synthesis in dealing with environmental pollution in general and dyes, difficultly- decomposed organic pollutants in particular, because selecting the combination of many microorganisms animals of likely higher laccase synthesis is not simple and requires much research effort This is the opportunity for research on purpose of finding a way to make up the processing technology to environmental pollution in general and water treatment technology of textile waste and environmental pollution of dioxin-containing herbicides in particular The results of the research on the ability of laccase need to continually complement the scientific base, a database of diverse levels of microbial strains laccase biosynthesis in general and on the geographical aspects different ecosystems in particular 1.1.2 Introduction of laccase-like In the course of its growth, the fungal strains beyond biosynthesis enzymes are protein in nature, they also have another ability of the biosynthesis of some biological agents with low molecular weights, the name of these substances depends on the source it is generated The agents are likely the same as laccase oxidation but chemical-physical characteristics are very different These compounds not have a protein structure but just are paragraphs peptides being highly oxidized A number of studies in Vietnam have discovered active substances generated from the strains of streptomycetes which was isolated from soil contaminated herbicides/dioxin at the Bien Hoa airbase, but detailed research to learn the characteristic -as well as the possibility of their application in this kind of decolorization dyes, decomposing the organic compound biodegradation is virtually published Thus, to distinguish those names and the term of the previous research and conveniently in the name of research of the thesis, the active ingredient will be called laccase-like 1.2 The waste water pollution characteristics and textile treatment technology This section outlines characteristics of textile waste water, the technology currently used for textile dyeing wastewater treatment The overview of the research on the application of laccase, laccase biosynthesis microbial treatment of decolored dyes 1.3 Pollution herbicides/dioxin in Vietnam and the treatment technology This section outlines the current state of pollution by dioxin-containing herbicides in the "hot spots" in Vietnam, stressing the current state of contamination in Bien Hoa airbase The technology has been on world statistics and Vietnam in dioxin polluted environment The synthesis research focus on using extracellular enzymes in general and laccase in particular in the degradation of organic compounds and difficultlydecomposed organic compounds CHAPTER SUBJECTS AND RESEARCH METHODS 2.1 Research subjects 2.1.1 Microbial strains Make sure the fungal strains collected and isolated at an altitude above 600 m from Ba Vi National forests in three regions and in Ba Trai around the foot area of Ba Vi Hanoi; The strains of streptomycetes are collected from the batch of biological treatment of herbicides/dioxin-contaminated soil in the area XĐ-1 Dong Nai, Bien Hoa airbase; The strains of streptomycetes are collected from the polluted soil in Southwest new pollution area (Pacer Ivy) Dong Nai, Bien Hoa airbase; Basidiomycetes strain FBD154 was isolated from Bidoup Nui Ba, Lam Dong and FNBLa1 filamentous fungus was isolated from rice straw in Ninh Binh in the same collection of research groups in the Biotechnology Institute 2.1.2 Examination object The herbicides 2.4-D, 2, 4, 5-T standard purity analysis (PA) of the Sigma and the toxic congener 2, 3, 7.8-TCDD; Polluted soil herbicides/dioxins were collected from two areas of Bien Hoa airbase, on toxic average of about 20,000 ng TEQ/kg; Some synthetic dyes in groups of azo and anthraquinone; The commercial active dyes are collected from the plant X20/General Department of Logistics 2.1.3 Cultivated mediums Gause M; potato extract (DCKT); PDA/B; Czapeck,; PDBDT; MEG; Vis; TSH1 medium 2.2 Research Methods Due to performance of many experiments to determine the characteristics, capabilities of laccase, laccae-like and synthetic microorganisms strains laccase, laccae-like in this kind of decolorization dyes and biodegradable herbicides/dioxin should experimental scheme described by the diagram in Figure 2.1 Figure 2.1 Experimental scheme diagram 2.2.1 Isolated and cultured of microorganisms 2.2.2 Classification of microorganisms 2.2.3 Biochemistry methode 2.2.4 Determination of decolorization ability 2.2.5 Determine the ability of decomposing herbicides/dioxin 2.3 Data processing methods Analysis data is processed by Microsolf excel/Microsoft office 10 software CHAPTER RESEARCH RESULTS AND DISCUSSION 3.1 Isolation, selection, and identification of fungi and ascomycete strains biosynthesis capable of laccase, laccase-like 3.1.1 Isolation, selection and identification of fungi strains at high laccase Have collected 45 fungal basidiomycetes in nature and in the ground of decayed wood at the items sample areas After four days, 22 strains of fungal has well-developed fibers, spread on the surface environment, fungi foam cotton fiber system is white, smooth and round red-brown on the creation of the environment containing the substances directive guaiacol Laccase activity in situ was identified shortly after the fungal samples were returned to the laboratory to be conducted The results obtained demonstrate the strains capable of biosynthesis enzymes in extracellular peroxidase (MnP, LiP) or called oxidoreductases (laccase) The results showed that there are about 16 samples of fungi collected in Ba Vi national forests for laccase biosynthesis Based on results of activity measurements of in situ laccase and growth capabilities Based on results of activity measurements of in situ laccase and the ability to grow fast, easy, FBV40 strains were selected to study the classification, enzyme properties, capable of decolorization dyes and biodegradable herbicides/dioxins Such a review on the characteristics of samples, the morphology of bacteria, spores and the sequence of the ITS1-5, 8S-ITS2 FBV40 fungus strains (already registered on the GenBank sequence with code MG243365) are gaining on the genus Rigidoporus and named Rigidoporus sp FBV40 Figure 3.2 Phylogenetic tree of FBV40 fungal strain 3.1.2 Isolation and classification of ascomycetes capable of growing on herbicides/dioxin medium and laccase-like biosynthesis From the polluted soil source, strains of actinomycetes were isolated, of which strains are (XKBHN1, XKBHN2) from Western land- and the other strains are XKBiR1 XKBiR2, XKBiR3, XKBiR4, XKBiR929, and XKBiR930 from the original soil (before processing the area Z1) contaminated herbicides containing dioxin in Bien Hoa airbase Based on the morphological characteristics of bacteria, spores and comparing the sequences of 16S rRNA encoding gene segments, XKBHN1, strain XKBiR929 was placed in the genus Streptomyces and named Streptomyces sp XKBHN1 and Streptomyces sp XKBiR929 11 About the durability of laccase purification, at pH Lac1 for active rest on 50% and in the pH value 3, active rest respectively in turn is 21 and 29% after h Meanwhile at pH Lac2, for active rest on 92% and at pH 3, 4, active rest respectively in turn is 54.55 and 78% after h As such, it can be seen that the pH conditions of laccase activity up isozyme from FBV40, especially vaccinations affect durability b) Influence of temperature on the activity and thermal stability Influence of temperature on the activity and the stability of the Lac 1, Lac from strain FBV40 has been implemented in about from 35 to 70oC, examination time to 140 minutes for Lac The temperature from 30 to 90oC and thermal stability has examinated from 35 to 70oC with time stretching to 180 minutes for Lac The results obtained both Lac1, Lac2, and reached highest activity at 60°C In the range of temperatures from 65 to 70°C reduced Lac1 active respectively in turn from 15 to 23% Lac2 reduce active from 20, 32 and 75% when the corresponding temperature is 70, 80 and 90°C The most stable Lac1 activity at 35oC when still 18% compared with the original activity after 140 minutes, while it reaches 92% Lac2 active and 90% at 30oC and 40oC after 180 minutes c) Specific substrates Lac1 and Lac2 can oxidize the organic compounds specificity of laccase such as ABTS, syringaldazine (Syrin), 26, DMP and guaiacol (Gua) The activity Lac with the organic chemicals 2.6-DMP, guaiancol and syringaldazine 12 versus ABTS is 76.5%, 0.54% and 0.13%, corresponding to Lac is 74% 0.3%, and 0.2% d) Influence of inhibitors and metal ions The effect of some particular inhibitors protein on the activity of Lac1, Lac has been studied The results are combined as follows: Comment on the level of being inhibition Inhibitors Lac Lac Was completely inhibited Was completely SDS at any concentration inhibited at any concentration 10 mM activity was 10 mM activity was completely inhibited; inhibited by 92%, mM activity was mM was inhibited by EDTA inhibited 65.5% and 58.4% and mM was mM activity was inhibited by 31.6% inhibited 28.5% 10 mM activity was completely inhibited; ClmM activity was inhibited 72%; mM inhibitory activity was 9.4% mM and 10 mM activity were completely inhibited; L-cystein mM inhibitory activity 54.5% 10 mM activity was inhibited 48%; mM Arginin activity was inhibited 36.5; mM activity was inhibited 28.5% Note: "-" not tested The influence of metal ions such as follows: Comment on the degree of being influence ion Lac Lac 13 ion Cu2+ Mg2+ Ni2+ Mn2+ Co2+ Fe2+ Comment on the degree of being influence Lac Lac Increased 109% activity at Increased 112% to the concentration mM 184.6% activity at the concentrations from 1.0 to mM, respectively Inhibitory activity at Inhibitory activity at all the all the concentrations concentrations and ranged and ranged from from 21.5 to 78.95%, 11.6% to 27%, respectively, from 0.5 mM to corresponding to 0.5 5.0 mM mM to 5.0 mM In 1.0 mM 100.6%, increase Inhibitory activity in in the concentration of the all the concentration inhibitor and the rest are and ranges from 12.8% ranged from 5.3% to 44.4% to 30% with the respectively 0.5 mM to concentration from 0.5 mM mM to mM Inhibitory activity in Inhibitory activity in all the all the concentration concentration and range and range from 11.6% from 21.1% to 35% with the to 18.9% concentration from 0.5 to 5.0 concentrations from mM 0.5 to 5.0 mM Cause inhibition of Inhibitory activity in all the activity in all the concentration and range concentration and from 15.5% to 35.2% range from 14.5% to concentrations from 0.5 to 32.3% from 0.5 mM 5.0 mM concentrations up to mM Inhibitory activity in Inhibitory activity in all the all the concentration concentration and range from and range from 95.9% 71.6% to 93.3% to 100% with the concentrations from 0.5 mM corresponding 14 Comment on the degree of being influence Lac Lac to mM concentrations from 0.5 mM to mM Inhibitory activity at Increase 107% and 128% of all the concentration and range from 16.2% Ca2+ activity at the concentration to 23.3% from 0.5 of 0.5 and mM mM concentrations up to mM The results obtained from this study showed that the ion influence of metal ions onto activity of laccase biosynthesis by FBV40 strain are similar to the research internationally published beforehand, however there exist differences in the extent of inhibition between Lac1 and Lac2 3.2.3.2 Kinetics of purified laccase Km and Vmax of Lac1 is respectively 0.3 µ M and 200.000 µM/min Meanwhile with Lac2 is 0.4 µM and Vmax is 10,000 µM/min 3.2.3.3 The Physical-chemical characterization of the crude laccase Research on the influence of environmental factors onto the raw laccase activity has been carried out Table 3.10 Factors affecting on the crude laccase of FBV40 Factors affecting Change laccase activity The influence of pH pH The highest activity pH The activity lost pH Active rest on 65% after 120 minutes The influence of temperature 30 to 40oC The highest activity Be inhibited in turn is 15% after 50 50oC, 60 to 70oC minutes at 160oC and 40% after 79 minutes at 60oC and up to 98% after 20 15 Factors affecting 30oC and 40oC Change laccase activity minutes at 70oC After 160 minutes of activity is almost not be inhibited Catalytic kinetics Km 0.5 µM Vmax 25,000 µM/phút The influence of inhibitors With mM severely inhibited completely SDS and mM were 79% inhibition With mM severely inhibited completely L-lysis and mM inhibits 86% With mM severely inhibited is 92% and EDTA mM were inhibited 10% Effects of mental ions Inhibition of activity in all concentrations Co2+ and ranged from 44.7% to 79.5% with a concentration of to 10 mM Inhibition of activity at all concentrations Mn2+ and ranged from 47.3% to 80.2% at concentrations of to 10 mM Inhibition of activity at all concentrations K+ and ranged from 41.7% to 80.7% with a concentration of to 10 mM Inhibition of activity at all concentrations Na+ and ranged from 75.5% to 94% at a concentration of to 10 mM Inhibition of activity at all concentrations Mg2+ and ranged from 13.4% to 79.1% with a concentration of to 10 mM Inhibition of activity at all concentrations Ca2+ and ranges from 41.5% to 68.5% with a concentration of to 10 mM Inhibition of activity at all concentrations Fe2+ and ranged from 97% to 97.9% with a concentration of to 10 mM 16 Factors affecting Change laccase activity Inhibition of activity at all concentrations Ni2+ and ranged from 2.7% to 88.3% with a concentration of to 10 mM Inhibition activity at all concentrations Cu2+ ranged from 48.1% to 76.2% with a concentration of to 10 mM The results obtained from the research shows that there exists similar ability but also big differences in the level of influence of the same element on crude laccase activity and purified laccase strains of FBV40 The research results showed that to apply purified laccase or crude one into environmental pollution disposal, overall and detailed studies should be needed to determine the factors and their influence levels; thereby to improve the efficiency of their application 3.2.3.4 Special chemical-physical properties of purified laccase-like Purified laccase-like has the highest activity at pH 1, rising fast and almost not being identified at pH The purified laccase-like activity of vaccine XKBiR929 hardly decreases when being boiled in the time up to hours Even, when fractional distillation takes off on the device filmed at a temperature of 800C and a pressure of 0.8 atm, active laccaselike of this strain still does not change As such, laccase-like created from XKBiR929 strain is very heat- durable Based on the first study, the influence of organic loading ability of oxidation of laccase-like, calculated the value of it with value Vmax is 142,857 µ M/min and Km is 0.43 µ M Compared with the value of laccase strains of fungi FBV40, the speed of response of the laccase-like is just lower than Lac1(200,000 µ 17 M/min) and about 14 times higher than Lac2 (10,000 µ M/min) and more than times higher than the crude laccase (25,000 µ M/min) With such a value of Vmax and Km obtained shows the very high oxygen capacity of laccase-like, especially in the very low value of pH It is a very notable feature of this thesis that the active substances are found in Actinobacteria The most powerful inhibitor aganst laccase-like activity is L-lys In concentrations 10 mM, active laccase-like rest 14%, followed by EDTA Active rest 66.7% and SDS having affected at least with laccase activity-like rest 83.6% Among the metal cation activity test with laccase-like, cation Fe2 + -induced inhibition of strong activity in all the concentration of a survey Compared with the effects of metal ions for laccase strains of FBV40, it shows that this is a very special properties of laccase-like race from XKBiR929 The results obtained by the thesis concerning the characteristics of the laccasee-like are actinomycetales created, which are also the first results obtained when surveying the basic characteristics of the protein and showed the outstanding characteristics of laccase-like 3.3 Decolorization dyes and digradation herbicides/dioxin 3.3.1 Decolorization dyes by laccase, laccase-like 3.3.1.1 Decolorization synthetic dye color by crude laccase Laccase of FBV40 strains has highly capability in the category of basic colour azo like NY1 and the basic color as NY7 anthraquinon as NY5 even if present and not present mediator This result is the first one of the ability to color sorts NY1, NY5 and fungi in the genus NY7 Rigidoporus was isolated from natural forests in Vietnam 18 3.3.1.2 Decolorization reactive dyes used in the military by crude laccase a) Capable of decolorization a number of reactive dyes Laccase of FBV40 strains shows decolorization ability some commercial reactive dyes used to dye a few army military sewing The ability to decolorate types best for dyes MN.FBN was 92.17% with dye, NN.SG was 91.84% after 3.5 h b) The selection of reactive dyes eliminated the highest decolorization Compare the capability of decolorization of crude laccase vaccine FBV40 with MN.FBN dye 92.67% achieved the highest, with the dye of NN.SG was 91.84% The MN.FBN decolorization fastness rate reached over 85% while with the dye of NN.SG just get over 17% during 0.25 h Especially without mediator, laccase activity reduced at 62.9% when decolorization MN.FBN dyes and 91.4% for NN.SG after 3.3 and hours The dye MN.FBN was, thus, selected to further study c) Capable of decolorization dye MN.FBN with crude laccase in the presence of D-glucose After 138 hours, with a concentration of 50 mg/L of the dye MN.FBN, the crude laccase FBV40 decolorated 99% and 97.6% in the presence of 0.1 g and 0.05 g D-glucose 3.3.1.3 Decolorization the active dye MN.FBN by Lac1 After 24 h, in the presence of 500 ViO µ M, Lac of FBV40 strains decolorated 91% color of MN.FBN with initial concentration of 50 mg/L The results clearly indicate the role and the possibility of very specific laccase strains FBV40 in loại màu dyes in general, and the commercial activity of the plant X20/General Logistics Bureau in particular 19 3.3.1.4 Decolorization the active dye MN.FBN by purified laccase-like At pH 1, purified laccase-like of vaccine XKBiR929 with the presence of fastening substance ViO decolorated 76.3 % MN FBN after 23 h But only after 5h, laccase-like decolorated 75.8% with the original dye concentration of 100 mg/L With the usage of the other CGK: HBT, its capability of decolorization reached 54.7% after 23 h, and hit 54.0% after 5h The role of the CGK ViO for laccase-like strains XKBiR929 in decolorization is not great (just add performance increase was 22%), with CGK HBT performance only increased the color type The results of the research have opened a new research direction to apply laccase-like in eliminating of organic compounds in low pH conditions 3.3.2 Decolorization reactive dyes by fungi FBV40 strain 3.3.2.1 The ability to decolorate reactive dyes used to dye fabric for the military FBV40 Strains are capable of decolorization MN FBN at the highest of 99% after days being cultured For MY dye color EG and MY BES efficiency decolorization reached a low level, just determined to be from to 8% Observe of fruiting bodies growing in MY.EG and MY.BES dye solutions showed color absorption on fruiting bodies, whereas MN.FBN dye did not see the adsorption, in contrast with the color dyes for MN FBN did not see the adsorption on From the above phenomenon can identify strains FBV40 type coloring dye, MN FBN by the mechanism of decomposition, metabolism 3.3.2.2 Decolorization dye MN FBN in different concentrations 20 After to cultured days FBV40 strain type color dyes for MN FBN has a concentration of 100 mg/L achieved from 52.7% to 97.3 %, while in the concentration of the dye 50 and 200 mg/L , the type of color performance reach respectively from 49.8 % to 94.3% and 35.2 % to 97.1 % As such, FBV40 fungus strains have the ability to sort the highest color in the dye concentration 100 mg/l 3.3.2.3 Decolorization dyes MN.FBN with the presence of Dglucose After day performance, efficency decolourization dyes for MN FBN by FBV40 have highly reached from 31 to 97%, while the environment has no D-glucose, then the performance is just 8% The obtained results of the thesis, together with the research of the author worldwide, D-glucose can be carbon source or is the CGK appropriate to the process of coloring dye, MN FBN by FBV40 fungus strains of highly effective 3.3.2.4 Decolorization dyes MN.FBN in the presence of other sugars With the use of the sugars as a source of carbon, the Dglucose reaches the highest effiency decolorization of 96%, reaching 90% lactose, mannose, gained 87% sacharose is 74%, while the template does not add the sugars, only reached 28% after days The research results were presented on again affirmed and clearly demonstrated the role of carbon sources suits in general as well as demonstrate the role of D-glucose in decolorization reactive dyes MN FBV40 strains of FBN 3.3.2.5 Decolorization dyes MN.FBN in the presence of the different nitrogen sources 21 (NH4)2SO4 in the FBV40 strains for cultivation medium, its performance of decolorization reaches 67%,3 is 57% KNO and NH4NO3 is 54% after days compared with cultured form for the certificate is 29% The results obtained by the dissertation shows, nitrogen source inorganic influence grew to the ability and they play a very important role in the process of laccase biosynthesis of fungal strains 3.3.3.Degradation herbicides/dioxin by laccase and biosynthesis of fungal laccase 3.3.3.1 Degradation herbicides/dioxin by crude laccase a) Degradation of soil extracts After 13 days of experiments capable of degrading 2.4-D and the 2,4,5-T respectively reached 65.46% and 85.71% in experimental formula in the presence of acetate buffer and mediator ViO b) Degradation of 2,4,5-T pure After 20 days, crude laccase strains of FBV40 are given the ability to degrade for 2, 4, 5-T at different rates in the range from 40.55 to 98.45% respectively with 2, 4, 5-T is in the form of experiment is 1.401 mg to 0.037 mg c) Degradation of 2,4-D and 2,4,5-T in contaminated soils After 27 days, 2.4-D in contaminated soils degraded 49.5% and 2,4,5-T degraded 37.71% of initial concentrations in the very highly contaminated soil 556.92 respectively and 981.88 ppm 3.3.3.2 Degradation herbicides/dioxins by fungi biosynthesis laccase a) Degradation 2,4,5-T in contaminated soil at Bien Hoa formed airbase 2,4,5-T in soil have very high rate in reaching 473.5 mg/kg, after days FBV40 strains cultured biodegrade by more than 7% corresponds to 36 mg/kg, if measurement processing 22 performance/day for mass experiments, the FBV40 were processed are 0, 045mg 2,4,5-T/day b) Degradation 2.4-D and the 2, 4, 5-T in polluted soil in Bien Hoa formed airbase in single and mixed fungal strains The performance of single FBV40 strains for 2.4-D, 2, 4, 5T in polluted soil in bien Hoa formed airbase in turn is 11.44 and 12.13% after 10 days processing with initial content in turn is 62.95 mg/kg and 63.5 mg/kg Also when using mixed strains of FBV40, FBD154 and FNBLa1 with 1:1:1 rate by volume then the degradation performance 2.4-D, 2,4,5-T increased significantly and in turn reached 36.06% and 56.08% c) Degradation congener 2,3,7,8-TCDD in single and mixed fungal strains After 10 days, single race and racial mixtures have the potential to biodegrade congener 2,3,7,8-TCDD with 44.68% performance when the original toxic was 2,000 ng-TEQ/L and mixed strains have a higher potential to 92.89% performance after 10 days of cultivation THE CONCLUSION Having selected strains of FBV40 from 22 strains isolated from Ba Vi national forests, Hanoi with the highest crude laccase activity being 107.708 U/l on the medium following TSH1 on cultured FBV40 strains are classified in the genus Rigidoporus and named Rigidoporus sp.FBV40 Isolation and Streptomyces, classify named two strains Streptomyces of Actinobacteria sp.XKBHN1 and Streptomyces sp.XKBiR929 laccase biosynthesis-like in the medium containing PAHs, 2.4-D, 2, 4, 5-T and soil extract 23 Having purified and identified characteristics of laccase of FBV40 strains including Lac1 and isozyme Lac2 that have the molecular mass of 55 kDa and 60 kDa, both of whose has different private properties After 24 h of interaction, Lac1 eliminated 91% decolorization commercial dye MN.FBN at concentrations of 50 mg/L Laccase-like purified from Streptomyces sp XKBiR929 was very thermal stable, even at 1000C Within h the activity does not change; affected by protein inhibitors such as L-lys, EDTA, SDS that is very different from real laccase; decolorization 76% MN.FBN dye in the initial concentration of 50 mg/L in pH condition in the presence of mediator ViO after h Efficiency decolorization synthesis dye by crude laccase of FBV40 in the concentration of 100 mg/L is 82% after 2,5 h, 85% after h for synthetic dye azo NY1, NY7 and 70 % after 48 hours for anthraquinone dye NY5 There is an inverse ratio between the rate relationship decrease laccase activity with the concentration of fastening in the process of decolorization synthetic dyes After 138 h in the presence of 0.1 g of D-glucose, crude laccase from Rigidoporus sp FBV40 has decolorated 99% MN.FBN dye and with the absence of D-glucose, just 44% After a day of implanting Rigidoporus sp FBV40 in medium containing 100 mg/L MN.FBN dye and 0.5 g D-glucose 97.2% of the color was removed The crude laccase of FBV40 degradated 40.55 to 98.45% of 2, 4, 5-T purity in the laboratory with the weight of 2.5 mg When concentrations of 2,4-D, 2,4,5-T up to 556.92 mg/kg and 24 981.89 mg/kg, after 27 days, crude laccase degraded 49.5 and 37.7% After to 10 days of cultured strains Rigidoporus sp.FBV40 biodegradable 11.44% (7.2 mg/kg) of 2.4-D herbicides with initial concentrations of 62.95 mg/kg and 12.13% (7.7 mg/kg) 2, 4, 5-T with concentration is 63.5 mg/kg Make sure the mixture of FBV40, FBD154 and FNBLa1 biodegradable 56.07% (35.29 mg/kg) of 2.4-D and 36.06% (22.89 mg/kg) 2, 4.5-T After days of cultivation, strains FBV40 is degradable 44.7 percent congener 2,3,7,8-TCDD with toxicity 2,000 initial ng-TEQ/L, while the mixture of races of the fungus is FBV40, FBD154 and FNBLa1 degraded 92.9% Recommendation Continue to more research to develop the capabilities of laccase, laccase-like and synthetic microorganisms strains laccase, laccase-like so as to gradually perfect the innovative technology in processing of organic pollutants hard degradation in general and herbicides containing dioxin in particular, contributing to completing the remedial program of chemical poisons remain saved after the war in Vietnam and the treatment of pollution to protect the environment NEWS CONTRIBUTIONS OF THE THESIS 1.The first study on laccase-like characteristics biosynthesized by Streptomyces sp XKBiR929 and its ability in decolorization of the commercial active dye MN.FBN; Demonstrate fungi strain Rigidoporus sp FBV40 is biosynthesized with crude laccase activity up to 107,708 U/L 25 and has high potential in decolorization of azo and anthraquinone dyes used in the military State that Rigidoporus sp FBV40 was also capable of degradation 2,4-D, 2,4,5-T herbicides and the most toxic congener of dioxins 2,3,7,8-TCDD Additionally, clarify and confirm that the mixture of different fungi strains (i.e Basidiomycetes and Ascomycetes) isolated from nature in Vietnam degradate not only 2.4-D, 2,4,5-T but also degradating 2,3,7,8-TCDD with higher efficiency than the use of the single strain LIST OF PUBLISHED WORKS Phung Khac Huy Chu, Đao Thi Ngoc Anh, Đinh Thi Thu Hang, Đang Thi Cam Ha, Isolation, classification and study of laccase-like biosynthesis possibility of some actinomyces in herbicides/dioxin contaminated soil of Bien Hoa airbase, Dong Nai, Province, Hue University Journal of Science, (2015), 110 (11): 29-41 Phung Khac Huy Chu, Nguyen Hai Van, Dang Thi Cam Ha, Purification and characterization of laccase involved in the decolourization of synthetic dyes and 2,3,7,8-TCDD congener degradation by the white rot fungus isolated from bavi forest of Vietnam, Vietnam Journal of Science and Technology, (2017), 55 (4C): 180-185 Phung Khac Huy Chu, Đao Thi Ngoc Anh, Dang Thi Cam Ha, Basidiomycetes fungus synthesis laccase that has decolouration abitily of active dyes used to dye fabric sewing ammunition uniform, Journal of Military Science and Technology, (12-2017), 52: 169-179 ... Then this laccase has been in the greater consideration than any other Therefore, the subject of study and research with the name "Research on the ability of decolorization of reactive dyes and. .. Performance evaluation of decolorization types of synthetic dyes, reactive dyes by laccase, laccase- like strain of microbes and laccase biosynthesis, laccase- like; Evaluate the biodegradable herbicides... characteristics of the protein and showed the outstanding characteristics of laccase- like 3.3 Decolorization dyes and digradation herbicides/dioxin 3.3.1 Decolorization dyes by laccase, laccase- like