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Yarrowia lipolytica strain and method thereof for synthesizing erythritol

A technology of Yarrowia lipolytica and yeast strains, which is applied in the biological field, can solve the problems of increasing erythritol purification cost, erythritol energy consumption, and deepening the color of fermentation medium, and achieve high synthesis of erythritol capacity, energy saving, and improvement in conversion rate

Active Publication Date: 2013-10-30
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] In addition, the fermentation of erythritol is a relatively energy-intensive process
The fermentation medium usually needs to be sterilized under high temperature and high pressure conditions (121°C for 20-40 minutes), and the Maillard reaction between glucose and nitrogen source will occur under this condition, which will darken the color of the fermentation medium and increase the concentration of erythritol. Purification cost

Method used

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  • Yarrowia lipolytica strain and method thereof for synthesizing erythritol
  • Yarrowia lipolytica strain and method thereof for synthesizing erythritol
  • Yarrowia lipolytica strain and method thereof for synthesizing erythritol

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1, the acquisition of Yarrowia lipolytica (Yarrowia lipolytica) BLC13

[0056] 1.1 Acquisition of synthetic erythritol yeast strains

[0057] Isolation of wild yeast strains that synthesize erythritol: The inventor collected 210 honeycomb samples from the mobile apiary in Wannian County, Jiangxi Province from April 30 to May 4, 2012, and brought them back to the experiment in sterilized 2ml centrifuge tubes chambers are separated. The honeycomb was rinsed with sterile water, and the obtained rinse solution was spread on the separation medium, and cultured at 30°C until colonies grew out. The composition of the separation medium is (g / L): fructose 450, yeast extract powder 20, peptone 10, agar 20, pH 5.5, and sterilized. After 7 days of cultivation, colonies with a diameter greater than 2.0mm were transferred to the fermentation medium to conduct the test of synthesizing erythritol. The composition of the fermentation medium is (g / L): glucose 280, yeast extr...

Embodiment 2

[0086] Embodiment 2, ferment and synthesize erythritol test in shake flask

[0087] 2.1 Experiment of Yarrowia lipolytica (Yarrowia lipolytica) BLC13 synthesizing erythritol from glucose in liquid medium I

[0088] Pick a Yarrowia lipolytica BLC13 yeast single colony (3-day bacterial age) and insert it into a 20ml test tube containing 3ml of liquid fermentation medium I and cultivate it at 30°C for 24 hours at 200 rpm. This 3ml nutrient solution is then all inserted into the 250ml shake flask with groove at the bottom containing 25ml liquid fermentation medium 1 to ferment and synthesize erythritol test (the groove at the bottom helps to increase dissolved oxygen). The composition of described liquid fermentation medium I is: glucose 200g / L, yeast extract powder 2g / L, diammonium hydrogen phosphate 1g / L, magnesium sulfate 0.2g / L, manganese sulfate monohydrate 0.01g / L, sulfuric acid pentahydrate Copper 0.005g / L, zinc chloride 0.002g / L, pH5.5. Ferment at 25°C and 200 rpm, take ...

Embodiment 3

[0100] Embodiment 3, feed-batch fermentation or continuous fermentation synthetic erythritol in fermentor

[0101] 3.1 Yarrowia lipolytica (Yarrowia lipolytica) BLC13 synthesizes erythritol from glucose test - fed-batch fermentation in 150 liter fermenter (a)

[0102] (1) Cultivation of slant seeds: Yarrowia lipolytica BLC13 was inoculated into an eggplant-shaped culture flask containing 70 ml of solid medium, and cultured in a biochemical incubator at 30°C until the cells covered the surface of the medium. The components of the solid medium are (g / L): 300 glucose, 10 yeast extract powder, 0.5 magnesium sulfate, 5 diammonium hydrogen phosphate, 10 corn steep liquor powder, 20 agar, pH 4.0, and sterilized at 121°C for 20 minutes.

[0103] (2) Wash the Yarrowia lipolytica BLC13 cells from the eggplant-shaped culture flask, transfer to a 1000-ml sterilized Erlenmeyer flask, and then transfer to a 150-liter fermentor containing 90 liters of fermentation medium. The fermentation ...

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Abstract

The invention discloses a yarrowia lipolytica strain and a method thereof for synthesizing erythritol. The yarrowia lipolytica strain is yarrowia lipolytica BLC13 CGMCC NO. 7326. The method for synthesizing the erythritol by the yarrowia lipolytica strain comprises the following steps of: taking a carbon source with an initial fermentation concentration of 100-400g / L, a nitrogen source with an initial fermentation concentration of 2-35g / L and inorganic salt as raw materials, treating for 30 minutes at 80-90 DEG C, cooling down and subsequently inoculating the yarrowia lipolytica strain, carrying out continuous fermentation or in-batch fed-batch fermentation under an aerobic condition, and after the fermentation is accomplished, purifying the erythritol from a fermentation liquid. The yarrowia lipolytica strain disclosed by the invention is used for synthesizing the erythritol from glucose, the conversion rate is high, and the Chinese erythritol standard is met.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Yarrowia lipolytica strain and a method for synthesizing erythritol. Background technique [0002] Erythritol (also known as butylene glycol) is a tetravalent polyol with a molecular weight of 122.1 Daltons. It exists in many fruits, vegetables and brewed foods (such as pears, glucose and other fruits and wines), but the content rare. Because erythritol has the advantages of zero calories, no dose limit, and pure biological fermentation preparation, erythritol is more widely used in food, and has been widely used in candies, baked goods, sugar-free drinks and other foods. [0003] For example: the Chinese invention patent whose application number is CN200710014055.8 discloses a method for making sandwich chocolate containing erythritol; the Chinese invention patent whose application number is CN201110420379. Sugar drink and its preparation method; the Chinese inventio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12P7/18C12R1/645
Inventor 程海荣吕霁烊王犇李德春邓子新
Owner SHANGHAI JIAO TONG UNIV
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