Screening And Compounding The Rapid Degradation Microorganisms Of Hyperaccumulation Organisms

For the scientific issues of fast volume reduction to enrichment plant collected from bioremediation which polluted by radioactive substances and heavy metal. The screening method was combined concentration gradient of uranium and associated heavy metal and cellulose, which w ere used for separating activated strains and also identified by molecular biology. This paper was discussed the related physiological and biochemical indexes and the optimal conditions to produce enzyme of these strains. The effect of strains could be improved by synergism and cooperative in the ultra-rich biomass disposal technologies. This research provided a theoretical and technical support for the disposal of excess enrichment plant. The results were as follows:(1)By the screening method of concentration gradient of uranium and associated heavy metal, and lignocellulose, the activitied strains were separated in which the Bacillus as a dominance. The Matching degree was 100% for C1 strain with Bacillus licheniformis BCRC 12826(DQ309323)and identified as Bacillus licheniformis; the matching degree was 100% for C3 with Bacillus subtilis subsp. Subtilis BCRC10255T(DQ309323)and identified as Bacillus subtilis subsp. Subtilis; the matching degree was 100% for C5 with Bacillus licheniformis BCRC 12826(DQ309323)and identified as Bacillus licheniformis; the matching degree was 100% for the C11 with Bacillus cereus WGPSB2(EF095752)and identified as Bacillus cereus; and the C16 was Trichoderma reesei CGMCCC3.3711.(2)By resistance screening of strains to uranium and associated heavy metal, the best culture condition was established that initial p H ranging from 5.5 to 8.0, inoculation quantity ranging from 2.0 % to 5.0 %, shaking speed ranging from 160 rpm to 200 rpm, and incubation temperature ranging from 28 to 37 ℃. Each strain could tolerate the stress effect of a certain concentration of uranium and associated heavy metal, which was different for the sensitivity to the above culture conditions, and the utilization of carbon source is also different.(3)The best condition of cellulase production of strain C1 was as follows: 1% of CMC- Na was used for carbon source of producing enzyme with fermentation. 2% of mixture of peptone and yeast powder was used as nitrogen source. The culture temperature was 39 ℃. The volume of liquid loaded was 70 m L. And the initial p H of medium was 8.The FPA and CMCA measured were: 143.67U/m L and 164.28U/m L respectively in the 4th day and 5th day.The best condition of cellulase production of strain C 3 was as follows: 1% of CMC- Na was used for carbon source of producing enzyme with fermentation. 1% peptone and 1% yeast powder was used as nitrogen source. The inoculation quantity was 3%.The culture temperature was 37 ℃. The volume of liquid loaded was 70 m L. And the initial p H of medium was 8. The FPA and CMCA measured were: 223.74U/m L and 257.67U/m L respectively in the 4th day and 5th day.The best condition of cellulase production of strain C 5 was as follows: 1% of CMC- Na was used for carbon source of producing enzyme with fermentation. 2% of mixture of peptone and yeast powder was used as nitrogen source. The culture temperature was 39 ℃. The volume of liquid loaded was 70 m L. And the initial p H of medium was 7.5.In these conditions, the capacity of cellulase production of bacillus licheniformis C5 was the strongest. The FPA and CMCA measured were: 215.74U/m L and 217.68U/m L respectively in the 3rd day and 5th day.The best condition of cellulase production of strain C 11 was as follows: 1% of CMC- Na was used for carbon source of producing enzyme with fermentation. 2% of mixture of peptone and yeast powder was used as nitrogen source. The culture temperature was 40 ℃. The volume of liquid loaded was 70 m L. And the initial p H of medium was 8. The FPA and CMCA measured were: 173.34U/m L and 159.57U/m L respectively in the 4th day and 3rd day.The best condition of cellulase production of strain C 16 was as follows: 1% of bran was used for carbon source of producing enzyme with fermentation. 2% of mixture of peptone and yeast powder was used as nitrogen source. The culture temperature was 30 ℃. The volume of liquid loaded was 70 m L. And the initial p H of medium was 5.5. The FPA and CMCA measured were: 235.89U/m L and 268.66U/m L respectively in the 5th day.(4)When collocations of different strains were used, the best culture condition was as follows: A2B3C3D3. The inoculation quantity of strain C1 and strain C5 mixture liquid was 4%. The inoculation quantity of bacillus subtilis was 6%. The inoculation quantity of waxy bacillus was 6%.The inoculation quantity of trichoderma reesei was 6%. Namely when the ratio of strain C1, C3, C5, C11, C16 was 1:3:1:3:3, the effect of degradation of straw was the best. The influence degree of Four factors in the experiment was,in order, B > D > C > A, namely the inoculation quantity of the bacillus subtilis has the greatest influence, followed by trichoderma reesei and waxy bacillus and bacillus licheniformis in order.