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Paenibacillussp and breeding method thereof as well as beta-glucanase produced from paenibacillussp and method for producing beta-glucanase

A Paenibacillus and glucanase technology, applied in the field of microorganisms, can solve the problems of low production level of β-glucan, insufficient source of strains, poor heat resistance, etc., and achieve easy observation and identification, easy operation, and high enzyme reaction Effect of temperature and temperature resistance

Active Publication Date: 2014-01-15
NANJING SOUTHERN ELEMENT BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on β-glucanase in my country started about 20 years later than that in foreign countries, and the production level of β-glucan still has some shortcomings, such as low production level, poor heat resistance, and insufficient source of strains.
Usually, β-glucanase-producing bacteria are screened from traditional substrates such as lichenin, pachyan, or yeast and mold cell walls, which limits the screening of new β-glucanase and its producing bacteria to a certain extent

Method used

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  • Paenibacillussp and breeding method thereof as well as beta-glucanase produced from paenibacillussp and method for producing beta-glucanase
  • Paenibacillussp and breeding method thereof as well as beta-glucanase produced from paenibacillussp and method for producing beta-glucanase
  • Paenibacillussp and breeding method thereof as well as beta-glucanase produced from paenibacillussp and method for producing beta-glucanase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Isolation and screening of β-glucanase producing bacteria S09 of the present invention

[0034] The rhizosphere soil samples of oats from the oat field in the suburbs of Nanjing were taken, added to the liquid screening medium for enrichment and cultured for 3 days, then the bacterial solution with a high degree of viscosity reduction was selected and diluted, and spread on the solid medium with Sola gum as the only carbon source. Cultured in a constant temperature incubator at 28°C. After 3 days, observe the growth of the colony, cover the medium with 2% CTAB solution, and observe the formation of the hydrolysis circle of Sola gum after 20 minutes. The Sola gum around the colony secreting β-glucanase was degraded and could not be combined with CTAB to become transparent, and the unhydrolyzed Sola gum combined with CTAB to form a white insoluble precipitate. The colony that formed the hydrolysis circle was reinoculated into the liquid medium, and cultured wi...

Embodiment 2

[0036] Embodiment 2: Paenibacillus S09 of the present invention ( Paenibacillus sp.S09) identification

[0037] a. Morphological characteristics: Observed after 2-4 days of culture on the inorganic salt sola gum solid medium at 28°C, the colony is light yellow and round, indicating smooth and moist, with neat edges.

[0038] b. Physiological and biochemical characteristics: It can effectively reduce nitrate, cannot degrade cellulose, grows very slowly in LB medium, but can grow rapidly after adding β-glucan. figure 1 b is the scanning electron microscope image of S09, which is short rod-shaped, without flagella, about 0.5 μm × 2.5 μm in size, and Gram staining is negative. The bacterial strain is aerobic, and the optimum temperature for growth and enzyme production is 28-30°C, and can also grow at 37°C, and the optimum pH is 6.0-7.0.

Embodiment 3

[0039] Embodiment 3: the preparation of Paenibacillus S09 fermentation crude enzyme liquid

[0040] Prepared seed medium: KH 2 PO 4 1‰, NaNO 3 2.5‰, CaCl 2 0.1‰, MgSO 4 0.15‰, FeSO 4 ·7H 2 O 0.01‰, solar rubber 3‰, H 2 O1000mL, pH6.5. After the culture was sterilized under high pressure at 121°C for 20 minutes, it was cooled to room temperature, and a single colony of S09 on the inoculated solid medium was picked and cultured with shaking at 28°C for 3 days to form a seed solution.

[0041] Prepare fermentation medium: 1% peptone, 0.5% yeast extract, 1% NaCl, 0.75% sola gum, pH6.5, sterilize the culture at 121°C for 20 minutes, cool to room temperature, and add 2% seed culture solution , 28°C, 220rpm, shaking culture for 30h, centrifuging at 7000rpm for 15min to remove the bacteria, collecting the supernatant to obtain a crude enzyme solution.

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Abstract

The invention discloses a high-yield beta-glucanase strain capable of effectively degrading salecan. The classification name of the high-yield beta-glucanase strain is paenibacillussp.S09, the collection registration number is CCTCC NO: M2012196, the collection date is May 30, 2012, the collection unit is China Center for Type Culture Collection (CCTCC) with the address of Collection Center of Wuhan University, Wuchang Luojiashan, Wuhan, Hubei province, and the registration number of 16SrRNA gene sequence of the beta-glucanase strain on Genbank is JQ945736. The invention also discloses a method for producing beta-glucanase by using the paenibacillussp.S09 and a crude enzyme preparation obtained according to the method. The enzyme preparation has high enzyme activity of laminarin enzyme and lichenase, and has wide application prospects in the fields of animal husbandry, brewing industry, biological control and the like.

Description

technical field [0001] The invention belongs to the technical field of microbes, and more specifically relates to a high-yield beta-glucanase strain, a crude enzyme fermented and cultured by the bacteria and a preparation method thereof. Background technique [0002] β-glucan is a naturally synthesized non-starch polysaccharide. As an important component of the cell wall, it widely exists in higher plants and fungi, and can also be secreted by bacteria to the outside of the cell to become an exopolysaccharide. β-glucans from different sources have different glycosidic bond binding methods and ratios. The main chain is mainly connected by b-1,3 glycosidic bonds, and usually contains b-1,2, b-1,4, b-1,6 glycosidically linked branched chains. The β-glucan extracted from brown algae and yeast is mainly β-1,3-glucan with β-1,6-branches, and the β-glucan in cereal crops is mainly β-1, 3-1,4-glucan. β-glucan present in wheat crops has become the main anti-nutritional factor in a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12Q1/04C12N9/24C12R1/01
Inventor 张建法程瑞徐淋香
Owner NANJING SOUTHERN ELEMENT BIOTECHNOLOGY CO LTD
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