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Preparation method of new natural abscisic acid

A natural abscisic acid and seed technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of glucose repression, high production cost, and low fermentation yield, so as to reduce production cost and increase acid production rate effect

Active Publication Date: 2011-07-20
江西新瑞丰生化股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies such as glucose repression, low fermentation yield and high production cost in the process of producing natural abscisic acid by microbial fermentation known in the prior art, the inventors conducted in-depth research and found a more Simple and easy to implement, the yield of abscisic acid is higher, and the improved process with lower production cost has thus completed the present invention

Method used

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  • Preparation method of new natural abscisic acid
  • Preparation method of new natural abscisic acid
  • Preparation method of new natural abscisic acid

Examples

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Effect test

Embodiment 1

[0174] Use 25 1000ml Erlenmeyer flasks, each with 300ml medium A, sterilize at 120°C, inoculate the activated abscisic acid high-yielding strain Botrytis cinerea TBC-10 CGMCC No.1889 spore suspension after cooling, and place at 25°C , shake flasks for 24-46 hours. Inoculate the cultured primary seed liquid into a 100 liter fermenter (secondary seed tank) with 50 liters of medium B inside by 13%-15% inoculum, and cultivate it with aeration and stirring at a temperature of 26°C-28°C for 24 -40 hours.

[0175] Use a 1-ton fermenter for the third-stage tank fermentation, and the medium C in the tank is about 500L. After sterilizing with conventional high-pressure hot steam sterilization, inoculate the seed liquid of the second-stage tank according to the inoculation amount of 10%-15%, and ferment with aeration and stirring for 20 - After 40 hours, feed medium C is intermittently fed, the intermittent time is to feed once every 10-15 hours, and the amount of each feed is 0.1%-0.5%...

Embodiment 2

[0182] Use 50 1000ml Erlenmeyer flasks, each with 300ml medium A, sterilize at 120°C, inoculate the activated abscisic acid high-yielding strain Botrytis cinerea TBC-10 CGMCC No.1889 spore suspension after cooling, and place at 25°C , shake flasks for 24-33 hours. Inoculate the cultured primary seed solution into a 200-liter fermenter (secondary seed tank) with 100 liters of medium B inside according to the inoculum size of 15%-18%, and cultivate it with aeration and stirring at a temperature of about 26°C for 24-30 Hour.

[0183] Use a 1-ton fermenter for the third-stage fermentation. The medium C in the tank is about 500L. After sterilizing with conventional high-pressure hot steam sterilization, inoculate the seed liquid of the second-stage tank with 20%-25% of the inoculation amount, and ferment with aeration and stirring for 15 - After 24 hours, feed medium C is fed intermittently, the intermittent time is 1-3 times every 20-28 hours, and the amount of each feed is 0.05%...

Embodiment 3

[0189]Use 30 1000ml Erlenmeyer flasks, each containing 300ml medium A, sterilize at 120°C, inoculate the activated abscisic acid high-yielding strain Botrytis cinerea TBI-9 CGMCC No.0500 spore suspension after cooling, place at 27°C, Shake flasks were incubated for 24-33 hours. Inoculate the cultured primary seed liquid into a 100 liter fermenter (secondary seed tank) with 50 liters of medium B inside by 15%-20% inoculation amount, and cultivate it with aeration and stirring at a temperature of about 26°C for 24-30 Hour.

[0190] Use a 1-ton fermenter for the third-stage fermentation. The medium C in the tank is about 500L. After sterilizing with conventional high-pressure hot steam sterilization, inoculate the seed liquid of the second-stage tank with 10%-15% of the inoculation amount, and ferment with aeration and stirring for 15 - After 24 hours, continuously feed medium C at a rate of 0.1-5 L / h until 10 hours before stopping fermentation (lower tank). Add inositol contin...

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Abstract

The invention relates to a method for preparing natural abscisic acid by adopting the technique of fed-batch of muscle alcohol to remove glucose repressor during fermentation of genetic modified bacterium TNC-10 with Botrytis fungi, which comprises the following steps: culturing the Botrytis TBC-10 bacterium in a first grade liquid culture medium; inoculating a cultured first grade seed liquid on a second liquid culture medium to culture; flowed-adding supplement material into a third grade liquid culture medium and muscle alcohol at the same time for the fermentation and culture; collecting the abscisic acid from the ferment culturing liquid. The invention adopts the genetic modified bacterium obtained from the genetic engineering, and cooperates with the method of fed-batch of the muscle alcohol for fermentation, artificially controls the carbon and nitrogen ratio in the fermantation system, removes the glucose repressor, improves the utilization rate of the thallus to other saccharide in the culture medium, enables the bacterial strain to generate the abscisic acid with high substrate conversion and product synthesis speed, improves the yield and production efficiency, greatly saves the energy comsumption and the raw material, reduces the production cost and simplifies the production process.

Description

1. Technical field [0001] The present invention relates to a method for preparing natural abscisic acid by adding inositol in the fermentation process to relieve glucose repression, and a new strain used in the method, wherein the strain is obtained by gene-directed mutagenesis Obtained by improving natural abscisic acid producing strains. 2. Background technology [0002] Abscisic acid (ABA) is one of the five major plant hormones found in the world. Natural abscisic acid has a strong regulatory activity on the growth and development of crops, can promote the maturity and development of fruits, cereals, and beans, can greatly improve their yield and quality, and can greatly enhance their cold resistance, drought resistance and resistance. Salt-alkali ability, so it has broad application prospects. At present, the research on abscisic acid in the basic field has reached the level of plant cells and genetic engineering. However, since the optical configuration of natural a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C07C59/90C12P7/42C12R1/645C12R1/66C12R1/80
Inventor 谭红雷宝良李志东周金燕杨杰
Owner 江西新瑞丰生化股份有限公司
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