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Sinorhizobium meliloti and method for applying same for fermenting to produce manganese peroxidase

A technology of manganese peroxidase and Sinorhizobium, which is applied in the field of Sinorhizobium melilotus strains, can solve the problems of no reports of Sinorhizobium melilotus, and achieve the effects of easy control of growth conditions, promotion of high yield, and low production cost

Active Publication Date: 2012-04-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still very few research reports on bacteria that can produce manganese peroxidase, and there is no report on Sinorhizobium melilotus in this regard.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Sinorhizobium melilotus ( Sinorhizobium meliloti ) CCTCC NO: M 2011318 Screening isolation and identification

[0024] 1. Sinorhizobium melilotus J09 ( Sinorhizobium meliloti J09) screening separation

[0025] Melilotus sinensis J09 strain was isolated from wood chips obtained from wood processing self-employed households in Hexing Village, Sanyu Town, Tongzhou District, Nantong City. Get 1 gram of sawdust into a 250 ml sterilized Erlenmeyer flask, add 100 ml of sterile aqueous solution containing inorganic salts with a pH value of 7.0 (the contained inorganic salts and their concentrations are: K 2 HPO 4 1 g / L; NaH 2 PO 4 1 g / L; (NH 4 ) 2 SO 4 0.5 g / L; MgSO 4 0.2 g / L; MnSO 4 0.02 g / L), cultured at 33°C and 150 rpm for 12 hours; take the above culture solution and dilute it to 10 -5 、10 -6 、10 -7 Take 100 microliters of dilutions of each concentration and spread on aniline blue plate (yeast extract 5 g / L, peptone 10 g / L, sodium chloride 5 g / L, anilin...

Embodiment 2

[0041] Example 2 Fermentation of Melilotus sinensis CCTCC NO: M 2011318 to produce manganese peroxidase

[0042] (1) Slant culture: Inoculate Sinorhizobium melilotus CCTCC NO: M 2011318 on the slant medium and culture at 33°C for 24 hours. The slant medium used was (unit: g / L): sodium chloride 5, yeast extract 5, peptone 10, agar 20, pH 7.0, sterilized at 115°C for 20 minutes.

[0043] (2) Seed culture: Use an inoculation loop to take a loop of the grown bacteria on the slant and insert it into the seed medium, and cultivate it at 33°C for 30 hours, and the shaker rotates at 150 rpm. The seed medium used is (g / L): 5 sodium chloride, 5 yeast extract, 10 peptone, pH 7.0; 30 ml seed medium per 250 ml Erlenmeyer flask, sterilized at 121°C for 30 minutes.

[0044] (3) Liquid fermentation culture: the fermentation medium used is (unit: g / L): lactose 15, peptone 5, magnesium sulfate heptahydrate 0.025, manganese sulfate 0.015, pH 7.0, sterilized at 121°C for 30 min. Put the culture...

Embodiment 3

[0045] Example 3 Fermentation of Melilotus sinensis CCTCC NO: M 2011318 to produce manganese peroxidase

[0046] The cultured seeds were inserted into a 500 ml Erlenmeyer flask, in which the fermentation medium was 50 ml, the inoculum size was 1 ml, and cultured at 36° C. for 72 hours with a shaker speed of 150 rpm. The fermentation medium used was (g / L): 30 lactose, 20 peptone, 0.025 magnesium sulfate heptahydrate, 0.015 manganese sulfate, pH 8.0, sterilized at 121°C for 30 min. Other conditions are the same as in Example 2, after the cultivation is completed, the activity of manganese peroxidase in the fermentation broth is 522 U / liter.

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Abstract

The invention relates to sinorhizobium meliloti and application thereof, which belong to the technical field of microbial fermentation. The sinorhizobium meliloti J09 is preserved in China Center for Type Culture Collection with the number of CCTCC NO: M 2011318. A method for applying the strain for producing manganese peroxidase through liquid fermentation comprises the following steps of: (1) slant culture; (2) seed culture; and (3) liquid fermentation culture: inoculating the cultured seed into a fermentation medium with the inoculation rate of 1 percent to 2 percent, and culturing at 32-36 DEG C for 96-136 hours with the rotating speed of a table concentrator of 150rpm. The sinorhizobium meliloti provided by the invention has the advantages that: the CCTCC NO: M 2011318 has the capability of producing the manganese peroxidase and higher biological nitrogen fixation activity. Since the strain is used for producing manganese peroxidase through liquid fermentation, the advantages that the production period is short, the production cost is low, the growth conditions are easy to control, and the like are achieved, and the method has potential for industrialization production. At the same time, the high-efficient biological nitrogen fixation activity shows that the strain has the capabilities of improving soil fertility and promoting high yield of crops.

Description

technical field [0001] A strain of Sinorhizobium melilotus and its fermentative method for producing manganese peroxidase, specifically a Sinorhizobium melilotus strain, a method for fermenting and producing manganese peroxidase using the strain, and its use in biological nitrogen fixation potential applications. The invention belongs to the technical field of microbial fermentation. Background technique [0002] Manganese peroxidase (MnP) was first isolated and purified from Phanerochaete chrysosporium. It can oxidize various aromatic compounds such as lignin. With the in-depth research on the cultivation, separation and purification, structure and function, and physical and chemical properties of MnP, its extremely high industrial application potential has also been continuously developed, such as in The degradation of organic pollutants, the decolorization of dyes, the biological bleaching of pulp, the treatment of agricultural waste, and the biodegradation of lignite a...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/08C05F11/08C12R1/41
Inventor 丁重阳杨晔顾正华李国辉彭林张梁石贵阳章克昌
Owner JIANGNAN UNIV
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