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Pectinase and preparation method thereof

A technology of pectinase and crude enzyme, which is applied in the field of pectinase and its preparation, can solve the problems of single pectinase preparation raw material, low enzyme activity, poor stability, etc., and achieve the purpose of providing juice yield and clarity, and high nutritional value , the effect of short process

Inactive Publication Date: 2018-02-16
GUANGXI AGRI VOCATIONAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a kind of pectinase and its preparation method in order to solve the problems of low utilization rate of main by-products in the existing banana deep processing process, single raw material for pectinase preparation, high cost, low enzyme activity and poor stability

Method used

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  • Pectinase and preparation method thereof
  • Pectinase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Aspergillus niger (CICC40493) uses banana peel residue solid fermentation to cultivate pectinase

[0032] step:

[0033] ① Banana peel residue pretreatment: Banana peel, sorting, drying (80~90°C, drying for 10~12 hours), smashing, passing through a 200~300 mesh sieve, for use, see the physical and chemical index control range of banana peel residue dry powder Table 1.

[0034] Total sugar (%)

40~45

Pectin (%)

14~16

protein(%)

2~3

Acid value(%)

2~3

Moisture (%)

4~4.5

Crude fat (%)

3.8~4.5

Ash content (%)

7~9

Other ingredients (%)

15~27.2

[0035] ②Strain preservation and seed expansion cultivation

[0036] CM0015 Chapeauer's medium was used for strain preservation and expanded culture. Culture conditions: Culture at 28°C for 100-120 hours at a constant temperature, and store the slant strains and the expansion culture strains in the triangular flask in a refrigerator at 0-5...

Embodiment 2

[0043] Embodiment 2: Extraction and separation and purification of pectinase

[0044] ①Concentration of crude enzyme extract

[0045] Pour the crude enzyme extract into an ultrafiltration cup for ultrafiltration concentration. The volume of the concentrated solution is about one-tenth of the original solution, and the molecular weight cut-off of the ultrafiltration membrane is about 10,000.

[0046] ② Ammonium sulfate precipitation

[0047] Put the concentrated pectin crude enzyme extract in an ice-water mixed bath, slowly add ammonium sulfate powder to the enzyme solution to a certain degree of saturation. During operation, it is necessary to stir while adding, and the stirring should be as slow and even as possible to avoid the generation of foam. Put it in the refrigerator and let it stand overnight, then freeze and centrifuge at 4000r at 4°C for 10 minutes to obtain the enzyme precipitate.

[0048] The initial concentration of ammonium sulfate fractional precipitation i...

Embodiment 3

[0060] Example 3: Enzymatic properties

[0061] ①Optimum pH

[0062] Buffers with different pH were prepared, and the enzyme activity of the crude enzyme extract was measured at 25°C to determine the optimum enzyme reaction pH.

[0063] The optimal reaction pH of this enzyme is 4. When pH<4, the enzyme activity is very low, and when the pH is in the range of 4.5-6.5, the enzyme activity decreases, but the change range is small. The suitable pH range of the enzyme is 4.5-6.5, and the test proves that the enzyme is suitable for reacting in a weak acid environment.

[0064] ②pH stability

[0065] The enzymes were placed in buffers with different pH, incubated at 37°C for 1 hour, and the loss of enzyme activity was measured. The enzyme activity at the most stable point was taken as 100%, and the enzyme activity at other pHs was expressed as the remaining enzyme activity.

[0066] Enzymes are most stable at pH 4.5. In the range of pH 4.5-6.0, the remaining enzyme activity was ...

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Abstract

The invention discloses pectinase and a preparation method thereof and belongs to the technical field of bio-fermentation engineering. The preparation method comprises the following steps: preparing banana peel pomace dry powder, activating strains, preparing seed liquid, carrying out solid fermentation culture, and preparing crude enzyme extracting liquid, wherein the step of activating the strains comprises that aspergillus niger is put into a czapek medium and culture is carried out for 100-120 hours at the temperature of 20-30 DEG C; and the solid fermentation culture comprises the steps of mixing the banana peel dry powder with wheat bran in proportion, adding deionized water, uniformly mixing, then adding ammonium sulfate, dipotassium phosphate and calcium chloride, stirring, charging, carrying out moist heat sterilization for 20-40 minutes at the temperature of 110-125 DEG C, inoculating spore suspension, and culturing for 60-72 hours. The preparation method disclosed by the invention solves the problem of utilization of a main byproduct in a banana deep processing process, preparation technology is simple, cost is low, pectinase activity is high, and juice yield and clarityof common fruit juice can be effectively improved.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation engineering, and in particular relates to a pectinase and a preparation method thereof. Background technique [0002] Banana belongs to Musa (Musaceae) and is one of the "four major fruits" in the world. Banana is one of the important tropical fruits in southern my country. Banana peel accounts for about 30-50% of its fruit weight. Studies have shown that banana peels contain a large amount of pectin, starch, sugars, cellulose, hemicellulose and lignin, as well as proteins, water-soluble sugars, unsaturated fatty acids, vitamins, inorganic salts, Ca, Mg, P and K and other nutrients. [0003] According to statistics, in the past ten years, China's banana production has increased from 6.5181 million tons in 2005 to 12.4663 million tons in 2015. According to the calculation that banana peels account for 35-40% of the total banana fruit, 4.3632-4.9865 million tons of bananas can be pr...

Claims

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Application Information

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IPC IPC(8): C12N9/18C12N9/24C12N9/88C12R1/685
CPCC12N9/18C12N9/2402C12N9/88C12Y301/01011C12Y302/01015C12Y402/02002
Inventor 韦璐唐婷赵琤陈智理李富宁
Owner GUANGXI AGRI VOCATIONAL COLLEGE
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