A kind of high amylase producing lactobacillus plantarum and application thereof
A technology of Lactobacillus plantarum and amylase, applied in the direction of Lactobacillus, application, bacteria used in food preparation, etc., can solve problems such as low efficiency
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Embodiment 1
[0038] Example 1: Isolation, screening and identification of Lactobacillus plantarum with high amylase production
[0039] 1. Preparation of culture medium:
[0040] (1) MRS liquid medium: peptone 10.0g / L, beef extract 8.0g / L, yeast extract 4.0g / L, glucose 20g / L, Tween 80 1.0ml / L, anhydrous dipotassium hydrogen phosphate 2.0 g / L, sodium acetate (3 crystal water) 5.0g / L, triammonium citrate 2.0g / L, magnesium sulfate (7 crystal water) 0.2g / L, manganese sulfate (4 crystal water) 0.05g / L. Adjust the pH to 6.0-6.4 and sterilize at 121°C for 20min.
[0041] (2) MRS solid medium: add agar powder 18.0g / L on the basis of the components of MRS liquid medium.
[0042] (3) MRS-1% starch liquid medium: peptone 10.0g / L, yeast extract 4.0g / L, Tween 80 1.0ml / L, anhydrous dipotassium hydrogen phosphate 2.0g / L, sodium acetate (3 crystals) water) 5.0g / L, diammonium citrate 2.0g / L, magnesium sulfate (7 crystal water) 0.58g / L, manganese sulfate (4 crystal water) 0.25g / L, soluble starch 10.0g / L....
Embodiment 2
[0061] Example 2: Enzymatic properties of Lactobacillus plantarum (CGMCC No. 14177)
[0062] 1. Determination of enzyme activity: DNS method (dinitrosalicylic acid method)
[0063] Preparation of standard curve: configure 0.1, 0.2, 0.3, 0.4 and 0.5 mg / mL maltose standard solutions, and measure the absorbance at a wavelength of 540 nm. The maltose standard curve was drawn with the concentration of maltose as the abscissa and the absorbance value as the ordinate.
[0064] Determination of enzyme activity curve:
[0065] 1% starch PBS buffer solution: respectively prepare 0.05mol / L disodium hydrogen phosphate and 0.05mol / L sodium dihydrogen phosphate, and mix the PBS buffer solution adjusted to pH 7 at 25°C. Take 1 g of starch, add a small amount of PBS buffer solution, heat and stir in a water bath to fully dissolve it, transfer it into a 100 mL volumetric flask, and dilute to 100 mL with PBS buffer solution for later use.
[0066] Take the cultured bacterial liquid, centrifu...
Embodiment 3
[0077] Example 3: Probiotic effect of Lactobacillus plantarum (CGMCC No. 14177)
[0078] 1. Study on acid resistance of strains
[0079]Adjust with hydrochloric acid, prepare MRS liquid medium with pH of 2, 3 and 7 respectively, take 150 μL of the above-mentioned medium and add it to a 96 microwell plate, and add 2 μL of bacterial solution cultivated for 18 hours at the same time, culture anaerobically at 37°C for 48 hours, and take samples every hour Determination of OD of culture medium 600 value, see Image 6 . The results showed that the strain could grow slowly under the condition of pH 3; but under the condition of pH 2, the strain could not grow.
[0080] 2. Research on Bile Salt Resistance of Bacterial Strains
[0081] MRS liquid medium with pH 7 and bile salt concentration (w / v) of 0%, 0.3% and 0.6% were prepared. Take 150 μL of the above-mentioned medium and add it to a 96-microwell plate, and add 2 μL of the bacterial solution cultured for 18 hours at the same ...
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