A strain of Paenibacillus and its application in the production of alkaline pectinase

A technology of Bacillus and pectinase, applied to a strain of Paenibacillus and its application in the production of alkaline pectinase, can solve the problems of low production of alkaline pectinase, and achieve beneficial preservation stability , the effect of reducing the accumulation of protease

Active Publication Date: 2015-07-29
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The production of alkaline pectinase of existing strains is often at a low level, even after fermentation optimization, the enzyme activity is only 1-10U / mL

Method used

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  • A strain of Paenibacillus and its application in the production of alkaline pectinase
  • A strain of Paenibacillus and its application in the production of alkaline pectinase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1, Screening, Identification and Preservation of Paenibacillus SJN-PL0602

[0024] 1. Screening of strains

[0025] 1. Collect rotten fruits and soil from Beijing orchard, Tianjin fruit beach, Tianjin saline-alkali land, etc., as samples for strain isolation.

[0026] 2. Dilute each sample with sterile water, and carry out enrichment culture in enrichment medium.

[0027] Enrichment medium (g / L): pectin 5-10, ammonium sulfate 2-5, sodium chloride 1, magnesium sulfate 1-3, dipotassium hydrogen phosphate 1, potassium dihydrogen phosphate 6, with 5mol / L NaOH Adjust the pH to 7.0-7.5. Enrichment culture conditions: 250mL shake flask (filling volume: 25mL), shaker speed 200r / min, culture at 37°C for 1d.

[0028] 3. Primary screening

[0029] The enrichment culture was carried out for 10 6 -10 9 After doubling dilution, take 0.1mL and spread it on the screening medium plate, culture it upside down at 37°C for 1-2 days, obtain a single colony, develop it with cet...

Embodiment 2

[0041] Embodiment 2, application Paenibacillus SJN-PL0602 produces alkaline pectinase

[0042] 1. Preparation of culture medium

[0043] Seed culture medium (pH6.8): Take 10g tryptone, 5g yeast extract and 5g sodium chloride, dissolve in water and dilute to 1L; sterilize at 121°C for 30min.

[0044] Fermentation medium (pH7.0): Take 5g peptone, 10g yeast extract, 3g glycerol, 3g pectin, 0.02g CaCl 2 , 2.31g of potassium dihydrogen phosphate and 12.54g of dipotassium hydrogen phosphate, dissolved in water and adjusted to 1L; sterilized at 121°C for 30min.

[0045] Two, the application of Paenibacillus SJN-PL0602 to produce alkaline pectinase

[0046]1. Inoculate Paenibacillus SJN-PL0602 in the seed medium (using 250mL shake flask, the liquid volume of the seed medium is 25mL), shake culture at 30°C (150r / min) to OD 600nm =1, that is, the seed solution.

[0047] 2. Transfer the seed solution in step 1 to 30mL fermentation medium (using 250mL shake flask) to obtain OD 600nm ...

Embodiment 3

[0064] Embodiment 3, application Paenibacillus SJN-PL0602 produces alkaline pectinase

[0065] 1. Preparation of culture medium

[0066] Seed culture medium (pH7.2): Take 24g tryptone, 12g yeast extract and 10g sodium chloride, dissolve in water and dilute to 1L; sterilize at 121°C for 30min.

[0067] Fermentation medium (pH7.5): Take 12g peptone, 24g yeast extract, 5g glycerol, 6g pectin, 0.1g CaCl 2 , 3g of potassium dihydrogen phosphate and 15g of dipotassium hydrogen phosphate, dissolved in water and adjusted to 1L; sterilize at 121°C for 30min.

[0068] Two, the application of Paenibacillus SJN-PL0602 to produce alkaline pectinase

[0069] 1. Inoculate Paenibacillus SJN-PL0602 in the seed medium (using 250mL shake flask, the liquid volume of the seed medium is 25mL), 37°C, shaking culture (200r / min) to OD 600nm =3, namely the seed solution.

[0070] 2. Transfer 1mL of the seed liquid from step 1 to 20mL fermentation medium (250mL shake flask), which is the initial fer...

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Abstract

The invention discloses paenibacillus and application thereof in production of an alkaline pectinase. The paenibacillus (Paenibacillus sp.) provided by the invention is named SJN-PL0602; and the preservation number is CGMCC No.5696. The invention also provides a method for producing alkaline pectinase. The method comprises the following steps of (1) inoculating the paenibacillus SJN-PL0602 to fermentation culture medium to obtain an initial fermentation system of which OD600nm is 0.05-0.15; and (2) carrying out following fermentation on the initial fermentation system: namely, firstly carrying out shake cultivation for 8-12 hours at 30-37 DEG C, and then carrying out shake cultivation for 36-40 hours at 22-26 DEG C to obtain the alkaline pectinase. By adopting the method disclosed by the invention, the enzyme activity of fermentation supernatant can achieve a high level; the paenibacillus has latent industrial value; and a foundation is established for subsequent industrial research.

Description

technical field [0001] The invention relates to a strain of Paenibacillus and its application in the production of alkaline pectinase. Background technique [0002] Alkaline pectinase (E.C.4.2.2.2) is a kind of pectin (a linear chain formed by D-galacturonic acid connected by α-1,4 glycosidic bonds) that can efficiently decompose pectin in plant tissues under alkaline conditions. The general term for enzymes of polymers). Alkaline pectinase is a kind of textile enzyme, which is mainly used in the bioscouring of cotton fabrics to remove pectin and other impurities in cotton fibers. Compared with traditional alkali treatment, enzyme treatment has the advantages of mild conditions, environmental friendliness, low energy consumption, and easy realization of clean production. significance. [0003] Because alkaline pectinase plays an important role in biorefining, many scholars at home and abroad are engaged in research work in this area. The production of alkaline pectinase ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/26C12P19/14C12R1/01
Inventor 宋江宁王辉林李小曼马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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