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Process for producing scyllo-inositol

A technology of scyllo-inositol and inositol, applied in the field of preparation of scyllo-inositol

Active Publication Date: 2006-11-22
HOKKO CHEM IND CO LTD (JP)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, in order to obtain scyllo-inositol from the complex, a large amount of organic solvent is required, so there is still room for improvement in terms of economical efficiency

Method used

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  • Process for producing scyllo-inositol
  • Process for producing scyllo-inositol
  • Process for producing scyllo-inositol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0342]

[0343] 3 liters of liquid medium containing 10.0% inositol, 1.0% yeast extract, and 1.0% sucrose were adjusted to pH 7.0 using 1N NaOH, and 100 ml were injected into 30 Erlenmeyer flasks with baffles each with a volume of 500 ml. Sterilize by autoclave. Each Erlenmeyer flask was inoculated with a slant culture 1 loop of Acetobacter AB10281FERM BP-10119, and cultured on a shaker at 27° C. for 5 days. After the cultivation, 250 ml of water was added to each Erlenmeyer flask, and the mixture was stirred for 1 hour on a shaker to dissolve the crystalline scyllo-inositol present in the culture solution. The culture solution was collected, centrifuged (8,000 rpm for 20 min), and the supernatant was obtained as the culture solution supernatant (10.2 L).

[0344] The culture supernatant was analyzed by high-speed liquid chromatography under the following conditions. As a result, 12.6 mg / ml scyllo-inositol (129 g, conversion rate 43%) was produced in the culture supernatan...

Embodiment 2

[0360]

[0361] A 40-liter liquid medium containing 10.0% inositol, 1.0% yeast extract, and 1.0% sucrose was introduced into a 50-liter fermenter, adjusted to pH 7.0 with 1N NaOH, and sterilized in an autoclave. Next, 400 ml of Acetobacter AB10281FERM BP-10119 cultured in a culture medium (erlenmeyer flask) with the same composition was cultured at 27° C. for 5 days at an aeration rate of 1 vvm and a rotational speed of 200 rpm. After culturing, 60 L of warm water at about 50° C. was added to about 40 L of culture solution taken out, and stirred for 1 hour to dissolve crystalline scyllo-inositol present in the culture solution. This culture solution was continuously centrifuged (8,000 rpm) to obtain a solution from which bacterial cells were removed as a culture sterilizing solution (about 100 L).

[0362] This culture sterilized solution was analyzed by high-speed liquid chromatography under the following conditions. As a result, 16.8 mg / ml scyllo-inositol (1.68 kg, conver...

Embodiment 3

[0367]

[0368] The 16S rRNA base sequences of four strains including three naturally isolated strains AB10285, AB10286, and AB10287, which have the ability to convert inositol into scyllo-inositol, and AB10281 bred from AB10253 were analyzed according to conventional methods. That is, after the genomic DNA was extracted from the cultured thalline, the corresponding DNA fragment was amplified by PCR with primers designed to amplify about 1.6kbp of 16SrRNA, and then the sequence related to about 1.3kbp was analyzed (Hokkaido System Science Corporation ). Based on the sequence results and compared with the database, related species were identified.

[0369] Table 2 shows the control results, homology, and the conversion rate to scyllo-inositol when cultured in the same manner as in Example 1.

[0370] system name

[0371] From this result, it was revealed that the four strains having the ability to convert inositol into scyllo-inositol can be roughly divided into ...

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Abstract

It is intended to provide a novel NAD<+>-independent myo-inositol 2-dehydrogenase which converts myo-inositol into scyllo-inosose in the absence of NAD<+>; a novel enzyme scyllo-inositol dehydrogenase which stereospecifically reduces scyllo-inosose into scyllo-inositol in the presence of NADH or NADPH; and a novel microorganism which belongs to the genus Acetobacter or Burkholderia and can convert myo-inositol into scyllo-inositol. By using these enzymes or the microorganism, scyllo-inositol is produced. Further, scyllo-inositol is purified by adding boric acid and a metal salt to a liquid mixture containing scyllo-inositol and a neutral saccharide other than scyllo-inositol to form a scyllo-inositol / boric acid complex, separating the complex from the liquid mixture, dissolving the thus separated complex in an acid to give an acidic solution or an acidic suspension and then purifying scyllo-inositol from the acidic solution or the acidic suspension.

Description

technical field [0001] The present invention relates to a process for the production of scyllo-inositol from inositol by microbial conversion. [0002] The invention also relates to novel NAD + Independent inositol 2-dehydrogenase and its preparation method. The present invention also relates to NAD + A screening method for microorganisms used to prepare blue crab inositol using the activity of independent inositol 2-dehydrogenase as an index. The invention further relates to the use of NAD + The preparation method of scyllo-inositol and scyllo-inositol of independent type inositol 2-dehydrogenase or the highly active strain of the enzyme. [0003] The invention also relates to a novel enzyme, i.e., scyllo-inositol dehydrogenase, and a method for preparing scyllo-inositol using the enzyme. In more detail, it relates to: a novel enzyme scyllo-inositol dehydrogenase that catalyzes the oxidation-reduction reaction between scyllo-inositol and scyllo-inositol, and stereospecif...

Claims

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Application Information

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IPC IPC(8): C12P7/18C07C35/16C12N1/20C12N1/21C12N9/04C12N15/53C12P7/26C12R1/02
Inventor 山口将宪北雄一森哲也神边健司友田明宏高桥笃市川稚子
Owner HOKKO CHEM IND CO LTD (JP)
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