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Culture medium for improving chitinase produced by streptomyces sampsonii KJ40 and chitinase bacteria-containing preparation and application

A technology of Streptomyces mulberry and chitinase, applied in the application, fungicides, enzymes and other directions, can solve the problems of accelerating product synthesis speed, affecting product synthesis, increasing temperature, etc., and achieving the effect of preventing poplar purple root disease

Inactive Publication Date: 2013-05-08
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From the perspective of enzyme reaction kinetics, the temperature rises, the reaction speed increases, the metabolism accelerates, and the product is produced in advance, but the bacteria are prone to aging and affect the product synthesis.
The size of the inoculum is related to the age of the strain and the growth and reproduction speed in the medium. If the inoculum is large, the seeds will be easier to adapt after entering the fermenter, thereby shortening the time required for the bacteria to multiply to the peak and speeding up the product synthesis speed; A large inoculation amount will usually cause the bacteria to grow too fast, resulting in a lack of nutrient matrix or insufficient dissolved oxygen, which is not conducive to fermentation; Mycelial clusters may form, leading to abnormal fermentation

Method used

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  • Culture medium for improving chitinase produced by streptomyces sampsonii KJ40 and chitinase bacteria-containing preparation and application
  • Culture medium for improving chitinase produced by streptomyces sampsonii KJ40 and chitinase bacteria-containing preparation and application
  • Culture medium for improving chitinase produced by streptomyces sampsonii KJ40 and chitinase bacteria-containing preparation and application

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Effect test

Embodiment 1

[0020] On the enzyme-producing basal medium, by adding carbon source, nitrogen source, and other single-factor experiments, the composition of the medium was determined, and an orthogonal experiment with 4 factors and 3 levels was designed to obtain the best enzyme-producing strain under the influence of 4 factors. The ratio is: colloidal chitin (solubility of colloidal chitin aqueous solution is 2%, w / V) 100ml / L, soybean cake powder 0.4%, K 2 HPO 4 1.0(g / L), KH 2 PO 4 0.6 (g / L). Among them, the content of nitrogen source has the greatest impact on enzyme production, followed by K 2 HPO 4 content, compared to substrate concentration and KH 2 PO 4 content had little effect on enzyme production.

[0021] 1.1 The relationship between culture time and enzyme production

[0022] Enzyme-producing basal medium (colloidal chitin 200ml, K 2 HPO 4 0.7g, KH 2 PO 4 0.3g, MgSO 4 ·7H 2 O0.5g, FeSO 4 ·7H 2 O0.01g, ZnSO 4 0.001g, distilled water 1000ml, pH7.0) as the initial ...

Embodiment 2

[0049] (1) Streptomyces sansoni seed production

[0050] 1. Strain activation: Transfer Streptomyces sanneri KJ40 slant strain to the plate (glucose 10g, peptone 5g, NaCl 5g, beef extract 5g, agar 20g, distilled water 1000ml, pH7.0-7.2), and culture at 25°C for 4 days.

[0051] 2. Preparation of Streptomyces sanneri seed solution: Prepare the prepared culture solution (colloidal chitin 200ml, K 2 HPO 4 0.7g, KH 2 PO 4 0.3g, MgSO 4 ·7H 2 O0.5g, FeSO 4 ·7H 2 O0.01g, ZnSO 4 0.001g, distilled water 1000ml, pH7.0) into 100ml Erlenmeyer flasks, 20ml per bottle, sterilized at 121°C for 30min, after cooling, add 1m concentration of 107 / ml spore suspension (Sanella bacteria) to each bottle, Then place it on a constant temperature shaker, and culture it at 26° C. with shaking at 140 r / min for 36 hours.

[0052] (2) Chitinase production and enzyme activity detection

[0053] 1. The best formula for producing enzymes: colloidal chitin (solubility of colloidal chitin aqueous solu...

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Abstract

The invention discloses a culture medium for improving chitinase produced by streptomyces sampsonii KJ40 and a chitinase bacteria-containing preparation and application. The culture medium is composed of 100ml / L of colloid chitin with solubility of 2%(w / V), 0.4wt% of soybean cake powder, 1.0(g / L) of K2HPO4, and 0.6(g / L) of KH2PO4. Compared with an enzyme production basic culture medium, the activity of the chitinase produced according to new formulation and fermentation condition disclosed by the invention is raised by 80%. The chitinase bacteria-containing preparation with 50-100 times (volume ratio) dilution can be used for performing root-irrigation on breadth of root system of poplar nursery stock with 100ml for each poplar, and is respectively applied for once in spring and summer so as to prevent violet root rot of the poplar.

Description

technical field [0001] The invention relates to a culture medium for improving the production of chitinase by Streptomyces sansoni KJ40, a preparation containing chitinase and its application. Background technique [0002] The production capacity of microbial strains can only be fully exerted under the optimum fermentation conditions. The production level of excellent microbial strains is restricted by many factors, including medium formula, initial pH, culture temperature, inoculation conditions, oxygen supply level, etc. Fermentation medium is the material basis for microbial growth, and its composition and ratio have a considerable impact on bacterial growth, anti-biomass fermentation units, refining process, and product quality. The improvement of fermentation medium is an important way to increase the level of fermentation. It must contain energy, elements and special nutrients that meet the needs of microorganisms. The types and concentrations of various compounds are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42A01N63/02A01P3/00C12R1/465
Inventor 朱天辉李姝江雷美艳张丽娜谯天敏
Owner SICHUAN AGRI UNIV
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