Study On The Physiological Activities Of Antimutagenic And Antitumor Of Korean Soybean Paste With Different Storage Period

ObjectiveSoybean paste,one kind of the traditional food of the Asia,not only taste delicious,but also nutritious,and contains a variety of functional ingredients which are good for human health,it has anti-oxidation,anti-aging,Anti-hypertension,Anti-hyperlipidemia,inhibiting tumor growth and lots of other physiological functions.Currently,researchers around the world are studying and looking forward to understand the physiologically active substances of fermented soybean paste more.As a representative of Korean food culture,the flavor of Korean soybean paste is unique.In recent years the study of traditional soybean paste has made many achievements,but the antimutagenic and antitumor activity research of Korean soybean paste with different storage period is still blank.Therefore,the results of this experiment may provide a theoretical basis for the further development of Korean soybean paste in nutrition,function,and for basic research and the promotion of cancer diet.Method1.This experiment adopts inoculated the Aspergillus sp.and Bacillus sp.to make Koji,and ferments under natural conditions to make Korean soybean paste.According to the national standard for basic component analysis and microbiological testing after mature fermentation(about 12 months)to verify its safety.2.Selecting the Korean soybean paste samples with different storage time for 0,1,3,6,9 and 12 months respectively,extracted with 70%ethanol,detected mutagenicity and anti-mutagenic effect by modified Ames test.Selecting MNNG and Trp-P-1-induced Salmonella typhimurium TA98 and TA100 strains for tested.And combined with the micronucleus test of bone marrow cells in mice to observe the anti-mutagenic effect of the Korean soybean paste extract during different storage period.3.Antitumor experiment was carried out at the same time,divided into two parts:in vitro antitumor test determined by MTT method,to detect the growth inhibition of Korean soybean paste extract on Hek-293,A549,AGS,MCF-7 and MDA-MB-231 cell lines;And in vivo antitumor effect uses S-180 tumor cell transplantation test,antitumor effect determined by intragastric gavage to mice with different storage period of Korean soybean paste extract.4.Soy isoflavones,the focus of research in recent years,the function is closely related with content,so this experiment selected commercial Korean soybean paste sold in Yanji City of Jilin Province based on different eating habits as testing samples,using high performance liquid chromatography(HPLC)method to determine the content of isoflavones in Korean soybean paste with different storage period(1-7 years).Result1.The moisture,protein,fat,minerals,dietary fiber and carbohydrate content of Korean soybean paste were 42.5%,19.5%,13.6%,19.5%,9.0%and 6.3%.Escherichia coli,Staphylococcus aureus.Vibrio parahaemolyticus and other pathogenic bacteria were not detected.2.In Ames test,the mutagenicity of Korean soybean paste extracts with different storage periods were negative;antimutagenic effect of Korean soybean paste extracts with different concentrations against TA98 and TA100 strains mutagenicity induced by mutagens(MNNG and Trp-P-1)were enhanced with the extension of storage time,the highest inhibition rate of concentration(200 μg/plate)were 90.87%,88.97%and 92.76%respectively.And there is a certain dose-time-response relationship around;there is significant inhibitory effect of Korean soybean paste extracts against mouse bone marrow micronucleus occurs induced by MNNG,in which micronucleus rate of the fermented extraction for 12 months was as low as 6.1±10.5/1000 cells,the the highest inhibition rate was up to 46.5%in the test.3.The antitumor test:extracts of Kroean soybean paste with different storage period showed weak growth inhibition effect on Hek-293 cells,the highest inhibition rate was 35%(1.00 mg/mL,stored for 12 months),but had strong inhibitory effect on A549、AGS、MCF-7 and MDA-MB-231 cells,and the inhibition rate gradually increased with the extension of storage time,the highest inhibition rate were 71.6%、76.7%、91.5%和 90.3%(1.00 mg/mL,stored for 12 months).Among them the extraction showed stronger inhibitory effect on breast cancer cells.The in vivo test showed significant inhibit effect on the growth transplantation tumor cell S-180 in mice,the inhibition rate was the highest with the storage time of 12 months,was 44.5%.4.The HPLC chromatograms of isoflavone content of commercially Korean soybean paste sold in Yanbian market stored for 1 to 7 years showed that aglycone isoflavone was much higher than glucoside isoflavone,and the trends was opposite with the extension of time.Aglycone isoflavone began to rise significantly from the fourth year of storage.ConclusionThe Korean soybean paste contains lower level of moisture and fat,but had a high content of protein,minerals and dietary fiber.And contains a variety of unsaturated fatty acids.The composition and proportion of amino acid is more reasonable.Harmful pathogenic microorganisms were not detected,in accordance with relevant national quality standards.Ames test showed the Korean soybean paste had no mutagenic,on the contrary,it showed stronger antimutagenic.Mouse bone marrow micronucleus test further validated the results.Antitumor test showed that the Korean soybean paste extraction can significantly inhibit the proliferation of A549,AGS,MCF-7 and MDA-MB-231,in particular showed the best inhibiting effect on breast cancer cell lines,while there is no obvious inhibitory effect on Hek-293.Meanwhile,the Korean soybean paste extraction has shown a strong inhibition effect on S-180 tumor growth transplanted into mice.With the extension of storage time,the aglycone isoflavone in soybean paste gradually conversed into a more physiologically active free isoflavone,as a typical representative of traditional fermented soybean paste Korean soybean paste,its isoflavone content were significantly higher than those fermented soybean products in other areas.