Isolation,identification And Physiology Property Analyses Of Microorganisms From Medicated Leaven And Chinese Gall Leaven During Their Production

Medicated Leaven,which is produced by microbial fermentation with flour, wheat bran, red bean, bitter almond, Artemisia annua, Xanthium sibirieum, Polygonum hydropiper, has a good effect on digestion and diarrhea. Chinese Gall Leaven, which is produced by microbial fermentation with gall, tea, wine yeast, has a good effect on dissolving phlegm and arresting cough. These two Chinese Traditional Medicines have been widely used in treating the diseases. However, during their production, there exist a variety of microorganisms of which the species and quantities cannot be ascertained. Meanwhile, some microorganisms can produce toxins, which cause adverse effect to patients’ health. In order to elucidate the beneficial microorganisms in Medicated Leaven and Chinese Gall Leaven made by traditional fermented methods, they were selected as the research objects in this paper. By using classical microbial isolation and purification methods, the strains in samples during production were obtained; by analyzing physiology biochemistry properties and combining their 16 SrRNA or 18 SrRNA gene sequence, the strains were prelimitary identified. Also, properties of isolated strains were investigated. The main contents and results of this paper are as follows.Six species of bacteria, one lactic acid bacteria, three species of yeasts and four species of filamentous fungi were isolated and identified in the first Medicated Leaven samples; five species of bacteria, one lactic acid bacteria, two species of yeasts and four species of filamentous fungi were isolated and identified in the second Medicated Leaven samples; five species of bacteria, one lactic acid bacteria, four species of filamentous fungi were isolated and identified in the third Medicated Leaven samples. By comparing the identified results, it was found that some strains may be the same ones and should be eliminated. There are eleven species of bacteria, two species of lactic acid bacteria, four species of yeasts and nine species of filamentous fungi in total. The bacteria were Pantoea agglomerans, Staphylococcus succinus, Enterobacter asburiae, Escherichia coli, Bacillus subtilis, Bacillus cereu, Acinetobacter calcoaceticus, Bacillus methylotrophicus, Enterobacter ludwigii, Cronobacter sakazakii and Enterococcus faecium1; the lactic acid bacteria were Pediococcus pentosaceus and Enterococcus faecium2; the yeasts were Hyphopichia burtonii, Debaryomyces hansenii, Pichia kudriavzevii and Saccharomycopsis fibuligera; the filamentous fungi were Mucor circinelloides, Mucor racemosus, Penicillium camemberti, Aspergillus candidus, Rhizopus oryzae, Lichtheimia corymbifera, Aspergillus fumigatus, Aspergillus oryzae and Penicillium phialosporum. However, Escherichia coli, Cronobacter sakazakii and Aspergillus fumigatus are known pathogenic microorganisms, they should be eliminated in later research.After investgating the microorganisms’ amount changes during their production, it was found that a majority of strains’ quantities presented upgrade firstly than descending latter tendency, and some strains’ quantities presented consistently declining tendency, and some strains appeared at the end of fermentation.By analyzing the enzyme properties, it was found that some strains showed digestive enzyme activities, such as Bacillus subtilis, Bacillus cereu, Bacillus methylotrophicus, Saccharomycopsis fibuligera, Penicillium camemberti, Aspergillus candidusucs and Penicillium phialosporum with protease and amylase activities, Lichtheimia corymbifera with protease activity, Hyphopichia burtonii and Aspergillus oryzae with amylase activities.For Chinese Gall Leaven samples, four species of bacteria and four species of filamentous fungi were isolated and identified. The bacteria were Bacillus oceanisediminis, Bacillus licheniformis, Bacillus firmus and Bacillus subtilis; the filamentous fungi were Aspergillus tubingensis, Aspergillus niger and two kinds of Penicillium chrysogenum. Moreover, Bacillus licheniformis, Bacillus subtilis and Aspergillus tubingensis showed protease and amylase activities; Penicillium chrysogenum1 showed protease activity; Penicillium chrysogenum2 showed amylase activity.