Screening Of Probiotics From Apple Pomace And Fermentation Production Of Microecologics

Apple pomace is the main by-product of apple processing industry, China’s annual processing of apple pomace produced about100million tons. Rational use of apple pomace is an important way to promote the efficient development of the apple processing industry and reduction of environmental pollution. Probiotics played a significant role in improving animal microbiological balance, preventing disease, promoting growth and purifying aquaculture environment. To develop new probiotics, was the effective way to achieve animal feeding safety, healthy, no residual contamination of non-anti-farming. In this paper, apple pomace as raw material, studied the microbial components of fermented apple pomace to get lactic acid bacteria and yeast. Fermentation technology of single bacteria and mixed bacteria for probiotics was optimized respectively, preliminarily determined the quality indicators of probiotics. The main results and conclusions were as follows:1. Microflora analysis of apple pomace and screening of fermentation strains10strains were isolated from the natural fermented apple pomace and were identified by the methods of morphology observation, physiological and biochemical characteristics tests and molecular biological techniques (ITS region,26S rRNA and16S rRNA gene sequence analysis). The results showed that10strains were:Aspergillus niger, Galactomyces geotrichum, Kluyveromyces marxianus, Aspergillus fumigatus, Lactococus lactis, Penicillium paneum, Candida tartarivorans, Lactobacillus plantarum and Issatchenkia orientalis. The principal microorganisms were made up of yeast and lactobacillus.5strains were studied by single factor experiment, based on the total number of bacterias after fermentation as an indicator. Ultimately determine the Lactobacillus casei and Kluyveromyces marxianus as the best fermentation strains.2. Optimizing the solid-state fermentation condition of single strainThe influence of different nitrogen sources on the solid-state fermentation of Lactobacillus casei was studied through the single factor experiment. Determine the fermentation medium was:Apple pomace50g/500mL, urea3%, potassium hydrogen phosphate0.2%, water material ratio of1.2:1. Optimization the fermentation conditions of Lactobacillus casei was:inoculun7.4%, fermentation temperature33℃and fermentation time79h. Under these conditions, the total experimental colonies reached6.235×109CFU/g.The influence of different nitrogen sources on the solid-state fermentation of Kluyveromyces marxianus was studied through the single factor experiment. Determine the fermentation medium was:Apple pomace40g/500mL, urea2%, potassium hydrogen phosphate0.3%, water material ratio of1.0:1. Optimization the fermentation conditions of Kluyveromyces marxianus was:inoculun6.6%, fermentation temperature28℃and fermentation time73h. Under these conditions, the total experimental colonies reached2.862x109CFU/g.3. Optimizing the solid-state fermentation condition of mixed strainsThe influence of different nitrogen sources on the mixed bacteria fermentation of apple pomace was studied through the single factor experiment. Determine the fermentation medium was:Apple pomace40g/500mL, urea3%, potassium hydrogen phosphate0.3%, water material ratio of1.2:1.The optimal fermentation conditions of mixed bacteria by the single factor and response surface methodology was:inoculun9%, vaccination than1:2(Lactobacillus:Yeast), fermentation temperature30℃and fermentation time92h. Under these conditions, the total experimental colonies reached8.347×109CFU/g.4. Probiotics preparation and analysis of quality stabilityFermented apple pomace using the optimized fermentation conditions to prepare probiotics, and to determine indexes of compound enzyme activity and polysaccharide. The study showed that enzyme activity of Lactobacillus casei was cellulase18.05u/g, xylanase35.47u/g, pectinase56.19u/g; Polysaccharide content was5.90%. Enzyme activity of Kluyveromyces marxianus was cellulase30.74u/g, xylanase58.30u/g, pectinase72.35u/g; Polysaccharide content was7.30%. Mixed strains fermentation of cellulase enzyme activity was49.21u/g, xylanase95.13u/g, pectinase126.30u/g; Polysaccharide content was10.80%. After stability test, bacteria amount remaining in108CFU/g.