The Research On Functional Genes Of Cellulase By Biosensor

Composting is considered as a useful process for the disposal of municipal and agricultural solid waste, in which enzymes play an important role.To monitor the trace of microbial community dynamics and the functional genes in composting system, which is favored the thorogh investigation of the microbial mechanism in waste composting process and efficient instruction of the microbial inoculation and composting technique renovation.Biosensing technology is a new approach for rapid, specific, real-time and on-line detection of analytes based on biometric identification. which has capacity for real-time and on-line detection in complex system in comparison with conventional analytical methods, and provides a new technical platform for the analysis and measurement of contaminants and various biological components or metabolites in relative degradation.The paper focuses on the trace of microbial community dynamics and the functional genes in composting system. Constructing a biosensing technology for the detection of contaminants and the dynamic changing of functional genes and metabolites in complex environment, which overcame the limitation of conventional analytical methods applied in environmental pollution control in the aspects of real-time and on-line detection, and etc.. Cellobiohydrolase (CBH) is a key enzyme in cellulose degradation. First, all the gene sequences of CBHII in Trichoderma reesei were found on NCBI, and the homology of all the sequences were analysised. According to results of the analysis, software design hybridization probes and PCR primers automatically.Multi-walled carbon nanotubes-macromolecule conductive polymer complex membrane and magnetic Au/Si/Fe core/shell nanoparticles were fabricated as a DNA carrier, which assembly with probes of CBHII in Trichoderma reesei. Then signal and target probes hybridization with the capture probes competitively, the amount of enzyme and carrier dosage, hybridization time and other hybridization conditions were optimized.DNA sensors based on multi-walled carbon nanotubes-poly (1-ethyl-(3-dimethylla-minopropyl) carbodiimide hydrochloride) complex membrane.Use of MWCNTs- PEDOT composite film can be more effective improved the electrode conductivity and enhanced sensitivity of the sensor. But with a carboxylic MWNTs not only with the NH2 of CBHII capture probe occurs cross-linked, but also with the HRP conjugate binding, there is interference in experiment, and requires further improvement.Use the advantages of nanoparticle moderate size, dispersion is easy to control, and synthesis of high biological compatibility and low cost and so on, magnetic Au/Si/Fe core/shell nanoparticles were synthesised by catalytic growth of Au NPs on the surface. An electrochemical nano DNA sensor based on the competitive hybridization recognition of CBHâ…¡was developed, with the lower detection limit of 10-13 M. Combining DNA extraction and polymerase chain reaction (PCR) technology, the cellulose and cellobiohydrolase gene extracted successfully. The sensor is useful for measured CBHII gene fragments from Trichoderma reesei genome extraction samples, to do further investigation. Key Words:Composting; Biosensor; Magnetic core/shell nanoparticles; Trichoderma...