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Production method of pullulan without pigment

A pullulan sugar and production method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of slow industrialization process and decreased fermentation efficiency

Inactive Publication Date: 2011-01-12
INST OF BIOPHARM OF SHANDONG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After 20 years of domestic research, great progress has been made, but there are still many problems. Among them, the conversion rate of melanin and substrate sugar produced in the fermentation process of Aureobasidium pullulans is the key problem restricting industrial production, and the current fermentation production of Pulu The time of blue sugar is generally about 120h, which leads to a decrease in fermentation efficiency. At present, it is difficult for many scientific research institutions at home and abroad to solve the above problems at the same time, so the industrialization process is quite slow.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Embodiment one: 30L fermenter fermentation and downstream treatment

[0010] Seed medium: sucrose 5%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5. Sterilize at 115°C for 20min.

[0011] The medium used for fermentation production is: sucrose 10%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O 0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5.

[0012] Seed liquid culture: inoculate the mutant strain of Aureobasidium pullulans obtained in 500mL Erlenmeyer flask with 1 / 5 seed medium, culture temperature 28±1℃, shaker speed 250r / min, culture 24h, seed concentration 2.0× 10 8 Individuals / mL, as the first-level seeds, put the first-level seeds into the 500mL Erlenmeyer flask with 1 / 5 of the seed medium according to the inoculation amount of the first-level seeds, the culture temperature is 28±1°C, and the shaker speed is 250r / min , cultivated for 24h as the secondary seed solu...

Embodiment 2

[0015] Embodiment two: 1m 3 Fermentation tank fermentation and downstream processing

[0016] Seed medium: sucrose 5%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5. Sterilize at 115°C for 20min.

[0017] The medium used for fermentation production is: glucose 10%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O 0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5.

[0018] Seed liquid culture: inoculate the mutant strain of Aureobasidium pullulans obtained in 500mL Erlenmeyer flask with 1 / 5 of the seed medium, culture temperature 28±1℃, shaker speed 250r / min, culture 24min, seed concentration 2.0× 10 8 Individuals / mL, as the first-level seeds, put the first-level seeds into the 500mL Erlenmeyer flask with 1 / 5 of the seed medium according to the inoculation amount of the first-level seeds, the culture temperature is 28±1°C, and the shaker speed is 250r / min , cultivated for 24h as the...

Embodiment 3

[0021] Embodiment three: 10m 3 Fermentation tank fermentation and downstream processing

[0022] Seed medium: sucrose 5%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5. Sterilize at 115°C for 20min.

[0023] The medium used for fermentation production is: sucrose 10%; yeast extract 0.2%; NaCl 0.1%; MgSO 4 ·7H 2 O 0.02%; K 2 HPO 4 ·3H 2 O 0.63%; (NH4) 2 SO 4 0.06%, initial pH 6.5.

[0024] Seed liquid culture: inoculate the mutant strain of Aureobasidium pullulans obtained in 500mL Erlenmeyer flask with 1 / 5 seed medium, culture temperature 28±1℃, shaker speed 250r / min, culture 24h, seed concentration 2.0× 10 8 Individuals / mL, as first-class seeds, put the first-class seeds into an 8L fermenter with 5L seed medium at a 2% inoculation amount, culture temperature 28±1°C, tank pressure 0.03Mpa, stirring speed 180r / min, ventilation rate 1L / min, cultivated for 24h as the secondary seed solution. Put ...

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Abstract

The invention relates to a fermentation production method of pullulan without pigment. Aureobasidium pullulans, which are used as the starting strain, are induced by microwaves to obtain the mutant strain with high-yield pullulan and little pigment secretion. The fermentation production method of the pullulan comprises the following steps: (1) preparing seeds and fermentation medium; (2) inoculating the activated strain to the seed culture medium according to the inoculum size of 2-10%, and culturing at 28+ / -1 DEG C for 24-36 hours, wherein the concentration of seeds is 0.8-2.5*108 / mL; (3) inoculating the seed liquid to the fermentation medium according to the inoculum size of 2-10%, pressurizing to 0.01-0.05 Mpa in a tank at 28+ / -1 DEG C, adjusting the stirring rate and ventilation amount at different fermentation stages and fermenting for 60-72 hours; and (4) after-treatment of fermentation liquid: using decoloration of the fermentation liquid, plate-and-frame filter, ceramic membrane ultrafiltration ultrafiltering for the sugar solution and the other methods to carrying out ultrafiltration and concentratuion, and performing spray drying to obtain the pullulan product.

Description

technical field [0001] The invention relates to a method for producing pullulan by microbial fermentation. Background technique [0002] Pullulan is a water-soluble microbial polysaccharide secreted by Aureobasidium pullulans. In 1939, R. Baner first discovered a viscous substance in the fermentation broth of Aureobasidium pullulans. That is pullulan sugar. In 1959, Bender et al. first characterized that the pullulan produced by the fermentation of Aureobasidium pullulans was composed of neutral glucose, which was composed of glucose combined with α-1,4-glycosidic bonds into maltotriose, and the two ends were separated by α-1 , 6-glycosidic bonds are repeatedly connected to form a polymer polysaccharide. It is a natural polysaccharide that is easily soluble in water, safe, non-toxic, edible, and low in calorific value. Therefore, it is often used as a packaging material for color retention, fragrance retention, freshness preservation, and anti-oxidation. Pullulan can be w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/10C12N15/01C12N13/00C12R1/645
Inventor 朱希强相茂功郭学平苏移山刘飞颜震陈勉张晓元候重文李海军李霞凌沛学
Owner INST OF BIOPHARM OF SHANDONG PROVINCE
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