Study On Enzymatic Activity Of Probiotics And Development Of Compound Enzyme Preparation

Probiotic enzyme is a kind of nutrient-rich and multi-effect healthy edible enzyme developed in recent years.It is deeply loved by consumers and also attracts the attention of researchers and the business community.With animal,plant or edible fungus as raw materials,it is prepared by probiotic fermentation containing specific biological active ingredients for human use with or without the addition of auxiliary materials.Among them,there are various kinds of biological active ingredients,including enzyme preparations,organic acids,amino acids,nucleotides,and functional sugars.Among them,enzyme preparations mainly include hydrolases,oxidoreductases,transferases,lyases,Isomerases and ligases,etc.At present,the study of superoxide dismutase?SOD?closely related to antioxidant activity in enzyme preparations is in the preliminary stage.This study mainly focuses on the research of probiotics fermenting fruits and vegetables to provide basis for the development of probiotic enzyme products with antioxidant activity.For this reason,the paper is based on Lactobacillus Plantarum,Lactobacillusparacasei,Bifidobacillusparacasei,Lactobacillus acidophilus and Lactobacillus rhamnosus as probiotics fermentation bacterium,using a variety of fruits and vegetables as raw materials for fermentation,conducting a pilot study of probiotics fermenting fruits and vegetables,and the results are obtained as follows:A preliminary study of SOD enzyme properties.By studying the effect of different concentrations of metal ions(Zn²⁺,Cu²⁺,Mn²⁺,and Fe²⁺)on SOD enzyme activity at different substrate concentrations?2.5mmol/L,3.0 mmol/L,3.5 mmol/L,4.0 mmol/L,4.5 mmol/L,5.0mmol/L,5.5 mmol/L,6.0 mmol/L,6.5 mmol/L,7.0 mmol/L,7.5mmol/L,8.0 mmol/L?,different temperatures?20?,25?,37?,42??,different p H values?2.0,2.5,3.0,3.5,4.0,4.5,5.0,5.5,6.0,7.0,and8.0?.we reach such conclusion that the SOD enzyme Km value was 3.57mmol/L;the SOD enzyme was at 37?.The activity was higher when the p H was 4.0-5.5.Zn²⁺,Cu²⁺,and Mn²⁺had little effect on SOD enzyme activity,and Fe²⁺had a significant inhibitory effect on SOD enzyme.Study on the Activity of SOD Enzyme Produced by Probiotics.Five strains including Lactobacillus bulgaricus,Lactobacillus paracasei,Lactobacillus plantarum,Lactobacillus rhamnosus,and Bifidobacterium lactis were inoculated into MRS liquid medium.The highest SOD activity was observed at 37?for 120hours.It was 60.71 U/m L,57.14U/m L,57.14 U/m L,39.29 U/m L and 17.86 U/m L.The activity of SOD enzyme was up to 71.43 U/m L during the incubation of five probiotics at 37?for 48 h.Research on probiotic fermentation of fruit.The impact of probiotics on fermented fruit was studied at different temperatures?20?,25?,37?,42??,different inoculations?5%,10%,15%,20%,30%?and different loadings?50%,60%,70,80%?.Through the observation of p H,acidity,number of viable cells,SOD enzyme activity and the morphology of the bacteria at 0 d,3 d,5 d,7 d,9 d,12 d,14 d,etc,the results showed that the SOD enzyme activity was 93.94 U/m L at90%fermentation time with 20%inoculation,70%loading and 25?temperature.At this time,the viable count was 1.41×10⁹/m L.Under this condition,the expansion test of probiotics fermented fruits was conducted.The raw materials were subjected to four treatments including beating and adding bacteria,beating without adding bacteria,cutting and adding bacteria,and cutting without adding bacteria.The fermentation period was 110 days.The results showed that:The SOD enzyme activity reached a maximum of 107.14 U/m L on the 3rd day of beating and adding bacteria,and the SOD enzyme activity reached a maximum of 42.35 U/m L on the 28th day of beating withou t adding bacteria.The maximum SOD enzyme activity was 105.36 U/m L in daytime,and the SOD activity reached a maximum of 59.66 U/m L on the 14th day of the fermentation without cutting bacteria.With the prolongation of fermentation time,the activity of SO D enzyme slowly decreased,and the enzyme was fermented to 110.At day time,the SOD enzyme activity was 6.92 U/m L in the samples that were excised only by cuttings,and no other SOD enzyme activity was detected.Research on probiotic fermentation of veget ables.The effects of different inoculums?5%,10%,15%,20%,and 30%?on the probiotic fermented vegetables were studied at different temperatures?20?,25?,37?,42??respectively,through observations of p H,acidity,number of viable cells,SOD enzyme activity,and cell morphology at the time of fermentation of 0 d,3 d,5 d,7 d,9 d,12 d and 14 d,the results were as follows:the SOD enzyme activity was higher at 95.75U/m L on the tenth day of fermentation with 20%inoculation volume,70%charge amount,and 25?temperature,and the number of viable cells was 1.32×10⁹/m L.Under this condition,the expansion test of probiotics-fermented vegetables was conducted.The raw materials were treated with four methods of beating and adding bacteria,beating without adding bacteria,cutting and adding bacteria,and cutting without adding bacteria.The fermentation period was 110 days.The results showed that:The SOD activity of beating and adding bacteria reached a maximum of 58.93 U/m L on the 5th day;the SOD activity of beating without adding bacteria reached a maximum value of 43.09U/m L on the 9th day;The activity of SOD enzyme reached its maximum value on the 7th day of fermentation,which was 101.16 U/m L and102.91 U/m L respectively.When it was fermented to 110 days,SOD enzyme activity was only 18.46 U/m L in the sample added to the cuttings only.No SOD enzyme detected.Preliminary study on compound enzyme preparation s.In the above selected conditions,the raw pulp of the probiotic fermented fruit was prepared,its SOD enzyme activity was 97.50 U/m L,the fermented pulp was beaten and sifted,the original pulp was added,5%lactose was added,and 1%was added.Glycine,p H 5,and 5%trehalose were added to 5 differently treated samples and freeze-dried at-50°C.and a vacuum degree of 10 Pa to prepare a SOD enzyme powder,and the freeze-dried powder was restored to puree.The SOD activity was measured to be 30.61 U/m L,50.10 U/m L,76.53 U/m L,88.78 U/m L,and97.33 U/m L respectively;the initial product of the enzyme preparation was stored at 4?and 25?respectively.The results showed that:the activity of SOD enzyme was slowly changed at the temperature of 4?.After 5 days,the activity of SOD enzyme was still high.With more trehalose added,the activity of enzyme activity was better.After 5 days,the activity of SOD enzyme was 73.96 U/m L.