Research On The Fermentation Technology Of Environmental Protection Enzymes In Thorn Pome Slag

Rosa roxburghii tratt,also named silk reeling flowers,send spring flowers,pear pomegranate,wooden pear etc.,is belong to the thorn pear family and the roses genus.In addition,it is the small deciduous and perennial shrub,and also the special wild fruit tree resources in Guizhou province.The fruit of Rosa roxburghii tratt contains rich Vitamin C,SOD,flavonoids and amino acids and has the extremely high nutrition,health care and medicine value.In recent years,with the rising of cultivated area of Rosa roxburghii tratt,the related industries also developed quickly,and many related products were developed.The large number of slags were generated during the processing progress of the thorn pear,however,there were many nutritional ingredients in the thorn pome slags which were disposed as the wastes.It was not only a wasting of resources,but it also polluted the natural environment.As a result,this study was focus on the preparation of the thorn pome slag enzymes.And also,the degradation effects of the thorn pome slag enzymes on the pesticides were explored,which could increase the utilization and additional value of the thorn pome slag.The thorn pome slag was regarded as a raw material,the fermented lactobacillus strain was isolated and identified,the processing progress of the environmental enzymes in the thorn pome slag was optimized.Furthermore,the degradation effects of the thorn pome slag enzymes on the pesticides were explored by studying the enzymatic activity of the thorn pome slag enzymes and related active ingredients.The research results were showed as followings:1.The screen of fermentation bacterial strain in the thorn pome slag enzyme:the five suspected strains of lactic acid bacteria were isolated from the thorn pome slag enzyme which was fermented at least 3 months.After that,according to the results of the related physiological and biochemical experiments and identification,the five bacterial strains isolated from the natural fermentation enzyme were detected by 16 S r DNA gene sequence analysis phylogenetic tree.The results showed that there were the highest homologies between S-3 bacterial strain and L.pentosus strain ATCC8041,CL-11 and L.pentosus strain Z2-2,S-9 and L.plantarum strain L009(LBF2)C03,S-4 and L.plantarum strain WS2,CL12 and L.plantarum strain XLP1733,and they had 99 % of affinities.The thorn pome slag contained many phenols and Vitamin Cs,and its p H was very low,so the bacterial were difficult to be survival.As a result,according to the detection of growth curves,ability to produce acid and activity of SOD enzyme,the lactobacillus CL12 was selected as the fermentation strain in the thorn pome slag enzyme.2.The optimum of the processing progress of environmental enzyme in the thorn pome slag: the fermentation processing progress of thorn pome slag enzyme was optimized by the response surface methodology with the lactobacillus CL12 and the commercial lactobacillus 21802.The effect of every fermentation factors including the inoculum size,the amount of brown sugar,initial p H,the solid-liquid ratio in fermentation,fermentation time and fermentation temperature on the enzyme activity of SOD and soluble acids were explored by the single factor and P-B experiment in the response surface methodology.In addition,the 4 significant factors including the inoculum size,the solid-liquid ratio in fermentation,fermentation time and fermentation temperature were screened by the P-B experiment,and the three-factors-four-levels response surface experiment was designed.The optimum processing conditions were: when the inoculum size of the lactobacillus plantarum21802 was 3.87%,the solid-liquid ratio in fermentation was 1:5.95,the fermentation time was 19.89 h,the fermentation temperature was 31.69 ?,the initial p H was 5.0and the amount of brown sugar was 4%,the SOD enzyme activity in the thorn pome slag enzyme was 402.127 U/ml and the amount of titratable acid was 2.58 g/kg;when the inoculum size of the lactobacillus plantarum CL12 was 4.75%,the solid-liquid ratio in fermentation was 1:5.56,the fermentation time was 18 h,the fermentation temperature was 36.04 ?,the initial p H was 5.0 and the amount of brown sugar was6 %,the SOD enzyme activity in the thorn pome slag enzyme was 404.882 U/ml and the amount of titratable acid was 4.16 g/kg;3.The related active ingredients and organic acids before and after fermentation of the thorn pome slag by the lactobacillus plantarum 21802 and lactobacillus plantarum CL12 were measured.And the results showed that there were no significant differences of cellulase activity,catalase activity and proteinase activity between the related active ingredients,the total phenol content before fermentation was lower than it after fermentation,the tannin content after fermentation was lower than it before fermentation.The results of organic acids and Vitamin C showed that the thorn pome slag enzyme contained the oxalic acid,lactic acid,citric acid,tartaric acid and acetic acid.Furthermore,these five organic acids contents were higher in the thorn pome slag enzyme which fermented by the lactobacillus plantarum CL12 than by the lactobacillus plantarum 21802.The Vitamin C content was no significant differences between these two lactobacillus plantarum.4.The degradation effect of the thorn pome slag enzyme on the chlorpyrifos and bifenthrin was investigated analysis by gas chromatograph.The thorn pome slag enzyme which fermented by the lactobacillus plantarum CL12 was mixed with these two kinds of pesticides(chlorpyrifos and bifenthrin)by ratio 1:1.After 24 h,the degradation ratios to bifenthrin and chlorpyrifos were 21.047% and 42.05%,respectively.After 96 h,the degradation ratios to bifenthrin and chlorpyrifos were64.65% and 44.08%,respectively.The thorn pome slag enzyme which fermented by the lactobacillus plantarum 21802 was mixed with these two kinds of pesticides(chlorpyrifos and bifenthrin)by ratio 1:1.After 24 h,the degradation ratios to bifenthrin and chlorpyrifos were 16.04 % and 35.1%,respectively.After 96 h,the degradation ratios to bifenthrin and chlorpyrifos were 47.726% and 35.1%,respectively.The thorn pome slag enzyme which fermented by the lactobacillus plantarum CL12 had a higher degradation rate to the chlorpyrifos and bifenthrin than the thorn pome slag enzyme which fermented by the lactobacillus plantarum 21802.And there was a significant degradation effect of the thorn pome slag enzyme on the bifenthrin after 96 h.The results were benefit to developing the degradation methods for pesticide residues.