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Method for preparing D-pantolactone by microbe mixed fermentation method

A pantolactone and mixed fermentation technology, which is applied in the field of microbial mixed fermentation to prepare D-pantolactone, can solve the problems of high total product yield and short hydrolysis time, and achieves short enzymatic conversion time and fermentation time. Short, efficient effect

Inactive Publication Date: 2009-03-25
JINAN DAHUA GUANGJI FARMING DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method has a short hydrolysis time, and although the hydrolysis rate is not high (about 30%), the unhydrolyzed L-pantolactone can be reused after racemization, and the total yield of the product is high

Method used

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  • Method for preparing D-pantolactone by microbe mixed fermentation method
  • Method for preparing D-pantolactone by microbe mixed fermentation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Slope culture: the medium is 160mL of potato juice (20%), 8g of sucrose, 8g of agar, 4g of peptone, 4g of yeast extract, about pH 7.0, set the volume to 800mL, sterilize at 121°C for 30 minutes, cool down and inoculate after sterilization , with Aspergillusterrus DH-069 as the small bacterium and Rhizopus oryzae QY-022 as the associated bacterium, the inoculum area of ​​the strains Aspergillus terreus and Rhizopus oryzae was 1cm 2 , Cultivated at 26°C for 3 to 6 days, used as slant activated seeds.

[0038] Enzyme production culture: the medium is 1% sucrose, 0.5% peptone, 0.5% yeast extract, 0.5% corn steep liquor, about pH 7.0, the liquid volume is 100mL / 500mL triangular flask, sterilized at 121°C for 30 minutes, after sterilization Cool and inoculate separately, and cultivate at 26°C for 3 to 5 days as a fermented enzyme liquid.

[0039] The shake flask experiment results are as follows (Table 1). It can be seen from Table 1 that the enzyme activity reached the hig...

Embodiment 2

[0044] The chemically synthesized DL-pantoacid lactone was used as the substrate, with 50mmol CaCl 2 The aqueous solution is prepared into a 30% concentration as the substrate: add the wet thallus prepared by the method of Example 1, the addition amount is 0.3U / g pantothenolactone, put it on a shaking table at 26°C, react at 160r / min, and take samples every 20min High-pressure liquid chromatography analysis and specific rotation determination, and the pH of the reaction solution was adjusted to 7.0 with concentrated ammonia water, and the enzymatic hydrolysis time was 5 hours. The mycelium filtered out after the reaction was finished was added with a new reaction substrate to continue the reaction. This was repeated and reused 8 times. The obtained enzymatic hydrolysis results are shown in Table 2. The results in Table 2 show that after repeated 8 times of enzymatic hydrolysis, the obtained enzyme conversion rate is about 40%, and the specific rotation of the product D-pantola...

Embodiment 3

[0048] According to example 1 method, ferment and produce enzyme, get wet mycelium 100g, press water: mycelia: glutaraldehyde=25: 3: 0.5, keep stirring 4~6 hours under 4 ℃, after immobilization finishes, weigh The immobilized mycelium that weighs about 100g left and right, with this immobilized mycelium, carry out enzymatic hydrolysis by the example 2 method. The results of repeated batch enzymatic hydrolysis for 10 times are shown in Table 3.

[0049] Table 3 Results of repeated batch enzyme transformation of immobilized cells

[0050] conversions 1 2 3 4 5 6 7 8 9 10 Conversion rates(%) 39.9 41.9 43.4 41.7 41.1 41.9 40.9 42.8 42.3 41.2

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Abstract

The invention relates to a method of D-pantolactone, in particular to a method taking chemically synthesized DL-pantolactone as a substrate for preparing the D-pantolactone by the mixed fermentation of microorganisms. The method adopts Aspergillus terrus DH-069-CGMCCNo.2498 and Rhizopus oryzae QY-022-CGMCCNo.2497 which are microbial flora for producing the hydrolase of the D-pantolactone as enzyme-producing strains for mixed fermentation. The method has the advantages that: the hydrolase of the D-pantolactone produced under the coordination of two strains is utilized to separate the DL-pantolactone, time for producing enzyme and fermentation is short (two days), enzymatic conversion time is also short (3 hours to 30 hours) and the enzyme can be reused for more than 8 times; one-time conversion rate of the enzyme is 38 percent to 56 percent; the total conversion of the substrate is 85 percent to 95 percent; the extraction yield of products is more than 60 percent; the major quality indexes of the D-pantolactone are as follows: specific rotation (Alpha)D<20> is minus 48 degrees to minus 51 degrees and optical purity is 99 percent e.e.

Description

technical field [0001] The invention relates to a method for D-pantoolactone, in particular to a method for preparing D-pantoolactone by using chemically synthesized DL-pantoolactone as a substrate and using microorganisms for mixed fermentation. Background technique [0002] D-pantoyl lactone (pantoyl lactone) is an important intermediate in the production of pantothenic acid, panthenol and pantethine. It reacts with β-alanine to obtain pantothenic acid. Pantothenic acid, Pantothenic acid, also known as pantothenic acid, vitamin B 5 or B 3 , is an important drug, food additive and feed additive. Pantothenic acid is a vitamin B group substance and a component of coenzyme A. It participates in metabolic processes such as fatty acid degradation and synthesis, citric acid cycle, choline acetylation, and antibody synthesis. The presence of pantothenic acid is conducive to the absorption and utilization of various nutrients . [0003] Because pantothenic acid is unstable to h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/04C12P39/00C12N1/14C12R1/66C12R1/845
Inventor 刘玉王友德庞志强刘洪玲乔刚
Owner JINAN DAHUA GUANGJI FARMING DEV
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