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Lactobacillus plantarum with high yield of amylase and applications of lactobacillus plantarum

A technology of Lactobacillus plantarum and amylase, applied in the directions of Lactobacillus, application, bacteria used in food preparation, etc., can solve problems such as low efficiency

Active Publication Date: 2018-06-01
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For enzymatic hydrolysis, B-type starch is more resistant to enzymatic digestion than A-type, and the surface of potato starch is smooth, without micropores and channels, so its enzymatic hydrolysis mode is "from outside to inside", and the efficiency is low

Method used

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  • Lactobacillus plantarum with high yield of amylase and applications of lactobacillus plantarum
  • Lactobacillus plantarum with high yield of amylase and applications of lactobacillus plantarum
  • Lactobacillus plantarum with high yield of amylase and applications of lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Isolation, screening and identification of plantarum lactobacillus with high amylase production

[0039] 1. Preparation 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 20 minutes.

[0041] (2) MRS solid medium: add agar powder 18.0 g / 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 crystallization 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 / ...

Embodiment 2

[0061] Embodiment 2: the enzymatic characteristic of plantarum lactobacillus (CGMCC No.14177)

[0062] 1. Determination of enzyme activity: DNS method (dinitrosalicylic acid method)

[0063] Preparation of the standard curve: configure maltose standard solutions of 0.1, 0.2, 0.3, 0.4 and 0.5 mg / mL, and measure the absorbance at a wavelength of 540 nm. With the maltose concentration as the abscissa and the absorbance as the ordinate, draw the maltose standard curve.

[0064] Determination of enzyme activity curve:

[0065] 1% starch PBS buffer solution: Prepare 0.05 mol / L disodium hydrogen phosphate and 0.05 mol / L sodium dihydrogen phosphate respectively, mix and adjust the PBS buffer solution to pH 7 at 25°C. Take 1g of starch, add a small amount of PBS buffer solution, heat and stir in a water bath to make it fully dissolved, then transfer it to a 100mL volumetric flask, dilute to 100mL with PBS buffer solution, and set aside.

[0066] Take the cultured bacterial solution,...

Embodiment 3

[0077] Example 3: The probiotic effect of Lactobacillus plantarum (CGMCC No.14177)

[0078] 1. Research on acid resistance of bacterial 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 ho...

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Abstract

The invention discloses lactobacillus plantarum with high yield of amylase. The strain is lactobacillus plantarum, is preserved in the China General Microbiological Culture Collection Center (CGMCC),and is assigned with the accession number of CGMCC No.14177, and the preservation date is May 19, 2017. The strain is separated from fermented old dough, is obtained through plate spreading, preliminary screening and secondary screening, and is identified through morphology, physiology and biochemistry and molecular biology. Culturing is carried out in an MRS culture medium (with the pH being 6) with 0.5% starch replacing 0.5% glucose for 21h at 31 DEG C, and the amylase activity of the fermented liquor is 15.89+ / -0.51 U / mL. The invention further discloses applications of the strain in solid fermentation of potato powder and applications of the strain in decomposition of starch. The strain can be used for fermentation of starchy raw materials including whole potato flour, and has the obvious degradation effect for starch.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a plant lactobacillus with high amylase production and application thereof. Background technique [0002] As a kind of biocatalyst, amylase has high specificity, acts on starch and undergoes a certain hydrolysis reaction, so it is widely used in starch grain processing, beverage production, pharmaceutical and textile industries. Studies have shown that α-amylase can reduce the viscosity of starch, make bread soft and enhance its extensibility, and β-amylase can hydrolyze starch to generate maltose, thereby delaying the aging caused by the interaction between starch and gluten, prolonging bread shelf life. Heteroamylase can decompose amylopectin into amylose. In industrial production, due to mild enzyme reaction, strong specificity and high catalytic efficiency, it is widely used. At the same time, the synergistic effect of various amylases has also attracted people's attention. [0...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L33/135C08B30/04C12R1/25
CPCA23L33/135C08B30/048A23V2002/00C12R2001/25C12N1/205A23V2400/169A23V2200/30
Inventor 张建华姚晓敏徐一涵周涛陈玉婧张立敏
Owner SHANGHAI JIAO TONG UNIV
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