Sinorhizobium meliloti strain and composition and application of sinorhizobium meliloti strain

A technology of Sinorhizobium and Sinorhizobium, applied in the direction of bacteria, microorganisms, microorganisms, etc., can solve the problems of inconsistent sample loading, low detection content, unfavorable detection, etc.

Active Publication Date: 2015-02-11
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

However, microbial detection takes a long time and has low sensitivity, which is not conducive to detection during fermentation; colorimetry is only suitable for the analysis of samples with relatively simple components such as drugs, but it has interference effects on samples with fermentation or complex components; Calculation of changes in vitamin B 12 Although the detection method is accurate and fast, the sample itself contains cobalt, which will cause a large error; due to the low detection limit of high-efficiency capillary electrophoresis, it will cause trace amounts of vitamin B in food, etc. 12 The detection has limitations; TLC has limitations in quantitative detection due to the inconsistent loading amount; high performance liquid chromatography can detect vitamin B in samples with low content and complex components 12 content, with the advantages of simplicity, speed and accuracy

Method used

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  • Sinorhizobium meliloti strain and composition and application of sinorhizobium meliloti strain
  • Sinorhizobium meliloti strain and composition and application of sinorhizobium meliloti strain
  • Sinorhizobium meliloti strain and composition and application of sinorhizobium meliloti strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Rhizobium Synthesis of Vitamin B 12 of fermentation:

[0057] Liquid seed medium: betaine (Tianjin Shilan Technology Co., Ltd.) 2 g / L, corn steep liquor (Tianjin Guanqian Chemical Distribution Department) 20 g / L, K 2 HPO 4 (Tianjin Guangfu Chemical Reagent) 1.65 g / L, sucrose (Sinopharm Holding Tianjin Co., Ltd.) 60 g / L, MgSO 4 (Tianjin Guangfu Chemical Reagent) 1.6 g / L, the rest is water, pH=6.8-7.2, sterilized at 110 ℃ for 15 min, and the liquid volume in a 250 ml Erlenmeyer flask is 30 ml.

[0058] Fermentation medium: sucrose (Sinopharm Holding Tianjin Co., Ltd.) 60 g / L, betaine (Tianjin Shilan Technology Co., Ltd.) 2 g / L, corn steep liquor (Tianjin Guanqian Chemical Distribution Department) 20 g / L, K 2 HPO 4 (Tianjin Guangfu Chemical Reagent) 1.65 g / L, MgSO 4 (Tianjin Guangfu Chemical Reagent) 1.6 g / L, ZnSO 4 (Tianjin Guangfu Chemical Reagent) 0.8 g / L, MnSO 4 (Tianjin Guangfu Chemical Reagent) 0.8 g / L, CoCl 2 (Tianjin Guangfu Chemical Reagent) 0.6 g / L...

Embodiment 2

[0061] Vitamin B 12 Extraction: Add 8% sodium nitrite and 2.5 ml of acetic acid to 10 ml of bacterial cell fermentation liquid, mix well, bathe in boiling water for 30 min, vitamin B in the bacterial cell 12 Transferred from intracellular to extracellular and converted into cyanocobalamin, resulting in vitamin B 12 of the mixture.

Embodiment 3

[0063] Determination of Vitamin B by High Performance Liquid Chromatography 12 content, the specific method is as follows:

[0064] Vitamin B 12 The pretreatment of mixed solution: the vitamin B that will be processed by embodiment 2 12 Let the mixture stand or centrifuge, take the supernatant, filter it through a filter membrane with a pore size of 0.22 μm, and the permeate is vitamin B 12 Sample to be tested.

[0065] Preparation of standard solution: Accurately weigh vitamin B 12 Standard substance (Sigma, purity ≥ 98%) 10 mg was prepared in a 25 ml volumetric flask with pure water to prepare a standard stock solution with a concentration of 400 mg / L. Then dilute the stock solution with pure water into standard solutions with concentrations of 40 mg / L, 80 mg / L, 120 mg / L, 160 mg / L, and 200 mg / L.

[0066] Qualitative and quantitative detection: under the same chromatographic conditions, vitamin B 12 The standard substance and the sample to be tested were determined by ...

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Abstract

The invention discloses a sinorhizobium meliloti strain capable of being applied to fermentation to generate vitamin B12, wherein the preservation number of the sinorhizobium meliloti strain is CGMCC No.9638. The invention also discloses a production and preparation method of vitamin 12 by the sinorhizobium meliloti strain in a fermentation manner. The method comprises a fermentation culture condition of the strain and an extraction method of the vitamin 12 in fermentation liquor; the method comprises the following steps: inoculating the strain on a solid culture medium into a triangular flask as a seed solution; inoculating the seed solution into a fermentation culture medium at 30 DEG C with 10% of inoculation amount; fermenting and cultivating at 200rpm for 200 hours, and then collecting the fermentation liquor; and adding a transforming agent to transform the fermentation liquor into cyano cobalt amine. The vitamin 12 is widely applied to the industries such as feeds, additives, medicines and cosmetics as a special factor in an adjusting organism growth process; and the requirement of the vitamin 12 rises year by year. The strain fermentation process is easy to control, so that the strain fermentation process is not affected by the factors such as seasons; the requirements on raw materials are simple; and fermentation production is developed into a main production mode.

Description

technical field [0001] The invention belongs to the technical fields of biological resources, microbial fermentation and macromolecular natural product biosynthesis. Involves a strain producing vitamin B 12 The strain, Sinorhizobium meliloti ( Sinorhizobium medicae ), and use the strain to ferment and prepare vitamin B 12 The method, more specifically a Sinorhizobium meliloti strain and its composition and application. Background technique [0002] Vitamin B 12 Also called cobalamin (Cobalamin), which belongs to corrin compounds, is currently the only vitamin compound containing metal ions, from vitamin B 12 It can be seen from the structure that its basic structure includes a corrin ring and a ligand. The central corrin ring is composed of 4 pyrroles connected to 1 cobalt atom, and the cobalt is chelated at the 4 pyrrole centers. Different types of ligands above the corrin ring form different types of cobalamins, generally four types can be formed: (1) Cyanocobalamin ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/42C12R1/01
CPCC12P19/42C12N1/205C12R2001/01
Inventor 张大伟李莎夏苗苗房欢周文娟郑平
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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