Difference Analysis Of Genome,Proteins And Metabolites Of Acidithiobacillus Ferrooxidans In The Bioleaching System Of Nano-realgar Particles

Recently,the researches of the realgar（As₄S₄）have been demonstrated that the agent has obvious anti-tumor effects.It is expected to be a candidate drug for the treatment of malignant tumors in the future.However,traditional processing methods still cannot solve the shortcomings of realgar with low solubility,high toxicity,poor bioavailability and difficulty in oral administration.In order to solve the above problem,bioleaching and nano-technology of realgar were used in the processing of realgar,which improved the solubility and bioavailability significant.Furthermore,the new technology of processing of realgar reduced the use of realgar and sovled the problem of easily oxidized and photochemical when realgar in nano-sized.In this study we have established the production process of preparation of nano realgar particles.The result demonstrated that when using high-energy ball mill to grind reagar pairicles the grind condition is as follow:the grinding speed at 600 r/min,ball material ratio at 1:5 and the grind time is 90 min.Under the grind condition above the size range of nano-realgar is all between 110-260 nm.Whatsmore,a process of using A.ferrooxidans DLC-5 to bioleach nano realgar particles was established.The results showed that when bioleach nano realgar particles20%of the inoculation of bacteria should add to the 9K medium contained 1 g/L Fe²⁺,keep the pH value at 1.5,maintain the temperature of bioleaching systerm at 35 ^oC,and 0.1 g of nano-realgar particles were added after adding bacteria for 20 hours in the medum.Under the bioleaching condition above the concentration of total arsenic in the realgar bioleaching solution is the highest.Physical and chemical properties of realgar particle and A.ferrooxidans DLC-5were measured by SEM、XRD、Raman and FT-IR before and after bioleaching of realgar.These results showed that As₂O₃ andβ-As₄S₄ were not detected in the bioleaching system.The infrared spectra of the bacteria showed that chemical groups on the cell surface are no obvious difference between the original strain and experimental strain.The result indicates that the quality of realgar and the activity bacterial has remained stable when using the A.ferrooxidans DLC-5 to bioleach nano-realgar.In biochemical activities analysis,the result indicated that the genome of A.ferrooxidans DLC-5 contained two plasmids and one circular chromosome.when searching the DNA database of the A.ferrooxidans we found that the DNA sequence of this genus of bacterias encodes a variety of proteins related to arsenic and sulfur metabolism.In proteomics studies,it demonstrated that the expression levels of arsenate reductase,iron-sulfur clusterin,and heat shock proteins in A.ferrooxidans DLC-5 strains have significantly changed.The result of differences in metabolites of A.ferrooxidans DLC-5 point out that the experimental group contained substances such as phenyl sulfate and sulfatide,and the same substance was not detected in the control strain.Therefore,we speculate that this phenomenon may be due to the bacteria metabolize realgar during bioleaching.The technology of using A.ferrooxidans DLC-5to bioleach nano realgars,and the multi-omics analysis of the bacteria not only can explain the mechanism of bioleaching processing,but also provide a more safted clinical candidate drugs in the future.