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A method for preparing (2s,3s)-2,3-butanediol and (3s)-acetoin from glucose

A technology of butanediol and glucose, applied in the field of biochemical industry, can solve the problems of restricted application, high cost, unsuitable for actual production, etc.

Active Publication Date: 2011-12-28
上海肆芃科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the cost of the substrate 2,3-butanediol and acetoin used in the above method is relatively high, which is not suitable for actual production, which restricts the application of the method
However, according to the retrieval results, there is no report on the method of simultaneously producing (2S, 3S)-2,3-butanediol and (3S)-acetoin in a two-step reaction using cheap glucose as a substrate.

Method used

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  • A method for preparing (2s,3s)-2,3-butanediol and (3s)-acetoin from glucose
  • A method for preparing (2s,3s)-2,3-butanediol and (3s)-acetoin from glucose
  • A method for preparing (2s,3s)-2,3-butanediol and (3s)-acetoin from glucose

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Experimental program
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Effect test

Embodiment 1

[0056] Embodiment 1: Preparation of Klebsiella pneumoniae CICC 10011 resting cell biocatalyst

[0057] (1) Slant culture: Streak Klebsiella pneumoniae CICC 10011 (purchased from China Industrial Microorganism Culture Collection Management Center) onto a solid medium slant containing 1.5% agar in a mass-volume ratio, and culture at 37°C for 14 hours .

[0058] (2) Seed cultivation: under aseptic conditions, use a sterile inoculation loop to pick a ring of bacteria sludge on the slope of step (1), inoculate into 50 ml of liquid medium, and incubate at 37° C. for 12 hours.

[0059] (3) Culture in a fermenter: Under sterile conditions, inoculate 5 liters of liquid culture medium with the inoculum size of 5% of the culture solution obtained in step (2), and cultivate at 37° C. for 16 hours.

[0060] (4) Collect the cells: Centrifuge the culture obtained in step (3) at 8,000 rpm for 10 minutes, wash the cells twice with pH 7.0 phosphate buffer, and then resuspend the cells in pH 7....

Embodiment 2

[0063] Embodiment 2: the mixture of (2S, 3S)-2,3-butanediol and meso-2,3-butanediol prepared by using the biocatalyst obtained in Example 1

[0064] Transformation experiment: the resulting biocatalyst, Klebsiella pneumoniae CICC 10011, had a cell concentration of 28 g dry cell weight / liter, an initial glucose concentration of 100 g / liter, and reacted at 37°C and pH 6.0. The reaction medium was for distilled water. The reaction was terminated after 8 hours to obtain a conversion solution containing a mixture of (2S,3S)-2,3-butanediol and meso-2,3-butanediol. The resulting conversion liquid was centrifuged at 8,000 rpm for 15 minutes to remove the added biocatalyst, and the concentration of (2S, 3S)-2,3-butanediol in the supernatant was measured by gas chromatography to be 3.2 grams per liter, meso- The 2,3-butanediol concentration was 40.7 g / l.

[0065] Collect the supernatant and carry out vacuum distillation at 50°C and 0.098 MPa, redissolve the fraction with a small amoun...

Embodiment 3

[0066] Embodiment 3: the mixture of (2S, 3S)-2,3-butanediol and meso-2,3-butanediol prepared by using the biocatalyst obtained in Example 1

[0067] Transformation experiment: the obtained biocatalyst, Klebsiella pneumoniae CICC 10011, was reacted at a cell concentration of 36 g dry cell weight / liter and an initial glucose concentration of 110 g / liter at 37° C. and pH 7.0. The reaction was terminated after 6 hours to obtain a conversion solution containing a mixture of (2S,3S)-2,3-butanediol and meso-2,3-butanediol. The resulting conversion liquid was centrifuged at 8,000 rpm for 15 minutes to remove the added biocatalyst, and the concentration of (2S, 3S)-2,3-butanediol in the supernatant was measured by gas chromatography to be 3.0 grams per liter, meso- The 2,3-butanediol concentration was 38.8 g / l.

[0068] Collect the supernatant and carry out vacuum distillation at 50°C and 0.060 MPa, and redissolve the fraction with a small amount of water to obtain a high-concentratio...

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Abstract

The invention discloses a method for prepared (2S,3S)-2,3-butanediol and (3S)-acetoin from glucose. The method comprises the following steps that glucose is converted into a mixture of (2S,3S)-2,3-butanediol and meso-2,3-butanediol by a Klebsiella pneumoma CICC 10011 resting cell biocatalyst; the mixture of (2S,3S)-2,3-butanediol and meso-2,3-butanediol is split by a bacillus subtilis ATCC 23857 resting cell biocatalyst; and (2S,3S)-2,3-butanediol and (3S)-acetoin are prepared simultaneously. The concentration of (2S,3S)-2,3-butanediol prepared by the method reaches 2.5g / L (wherein the optical purity is 96.9%). The concentration of (3S)-acetoin prepared by the method reaches 29.3g / L (wherein e.e. is 96.2%). Therefore, the method has great industrial application prospects.

Description

technical field [0001] The invention provides a method for preparing (2S, 3S)-2,3-butanediol and (3S)-acetoin from glucose, which belongs to the technical field of biochemical industry. Background technique [0002] 2,3-butanediol contains 3 isomers (2R,3R)-2,3-butanediol, (2S,3S)-2,3-butanediol and meso-2,3-butanediol . Acetoin contains two enantiomers (3R)-acetoin and (3S)-acetoin. The various isomers of 2,3-butanediol and acetoin are important raw materials in asymmetric synthesis and are pharmaceutical intermediates with great application potential. [0003] The current methods for producing chiral 2,3-butanediol and acetoin include chemical and biological methods. Among them, the reaction conditions of the chemical method are complex, difficult to control, high in cost, and the optical purity of the product is very low. The biological method has the advantages of various reaction types, strong reaction specificity, mild temperature and pressure conditions, environme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12P7/26C12R1/22C12R1/125
Inventor 许平马翠卿刘振
Owner 上海肆芃科技有限公司
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