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Method for Preparing D-Psicose Using Microorganisms of Genus Kaistia

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Method for Preparing D-Psicose Using Microorganisms of Genus Kaistia (19) TZZ¥¥¥ T (11) EP 3 543 345 A2 (12) EUROPEAN PATENT APPLICATION published in accordance with Art. 153(4) EPC (43) Date of publication: (51) Int Cl.: 25.09.2019 Bulletin 2019/39 C12P 19/02 (2006.01) C12N 1/20 (2006.01) C12R 1/01 (2006.01) (21) Application number: 17872289.8 (86) International application number: (22) Date of filing: 15.11.2017 PCT/KR2017/012971 (87) International publication number: WO 2018/093154 (24.05.2018 Gazette 2018/21) (84) Designated Contracting States: • LEE, Young Mi AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Suwon-si GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO Gyeonggi-do 16516 (KR) PL PT RO RS SE SI SK SM TR • KIM, Yang Hee Designated Extension States: Suwon-si BA ME Gyeonggi-do 16507 (KR) Designated Validation States: • KIM, Seong Bo MA MD Seongnam-si Gyeonggi-do 13567 (KR) (30) Priority: 16.11.2016 KR 20160152948 • PARK, Seung Won Yongin-si (71) Applicant: CJ Cheiljedang Corporation Gyeonggi-do 16894 (KR) Seoul 04560 (KR) • CHO, Seong Jun Seoul 04971 (KR) (72) Inventors: • KIM, Su Jin (74) Representative: Cabinet Beau de Loménie Suwon-si 158, rue de l’Université Gyeonggi-do 16500 (KR) 75340 Paris Cedex 07 (FR) (54) METHOD FOR PREPARING D-PSICOSE USING MICROORGANISMS OF GENUS KAISTIA (57) Provided are a composition for preparing D-psicose comprising a microorganism of the genus Kaistia, and a method for preparing D-psicose using the same. EP 3 543 345 A2 Printed by Jouve, 75001 PARIS (FR) EP 3 543 345 A2 Description TECHNICAL FIELD 5 [0001] The following disclosure relates to a method for preparing a D-psicose using a microorganism of the genus Kaistia. BACKGROUND 10 [0002] D-psicose (hereinafter referred to as "psicose") is a monosaccharide known as a rare sugar present in the natural world in a very small amount. It has almost zero calorie while having nearly 70% sweetness of sugar and has received a lot of attention as a new food ingredient due to its functionalities such as inhibition of blood glucose, and inhibition of lipid synthesis, etc. [0003] Due to these characteristics, psicose is considered to be used as a sweetener substitute for sugar in various 15 foods. However, there is an increasing need for a method for efficiently preparing psicose since it exists in a very small amount in the natural world. [0004] A known method for preparing psicose comprises a method for utilizing catalysis of molybdate ions (Bilik, V., Tihlarik, K., 1973, Reaction of Saccharides Catalyzed by Molybdate Ions. IX. Epimerization of Ketohexoses. Chem. Zvesti. 28:106-109), A Chemical Method for Preparing Psicose from D-Fructose by Heating Ethanol and Triethylamine 20 Together (Doner, L.W., 1979, Isomerization of D-Fructose by Base: Liquid-Chromatographic Evaluation and The Isolation of D-Psicose. Carbohydr. Res. 70:209-216), and A Biological Method for Preparing Psicose from D-Fructose Using a Microorganism that Produces D-Psicose 3-Epimerase (Korean Patent Laid-Open Publication No. 10-2011-0035805). Preparation of psicose by the chemical method has problems in that a large amount of byproducts occur, and thus, it is required to perform a complicated purification. Further, the biological method also has problems in that thermal stability 25 is not high and the preparation cost is high, and thus there is a need to provide a new microorganism being able to produce psicose from D-fructose. [0005] Under these circumstances, the present inventors have made a lot of effort to develop a new microorganism being able to produce psicose, and as a result, confirmed that when the microorganism of the genus Kaistia was used, psicose can be produced from D-fructose and completed the present invention. 30 TECHNICAL PROBLEM [0006] An embodiment of the present invention is directed to providing a composition for preparing psicose comprising a microorganism of the genus Kaistia. 35 [0007] Another embodiment of the present invention is directed to providing a method for preparing psicose comprising: contacting the microorganism of the genus Kaistia or the composition for preparing psicose comprising the microorganism of the genus Kaistia with D-fructose. [0008] Another embodiment of the present invention is directed to providing a microorganism of the genusKaistia useful for preparation of psicose. 40 TECHNICAL SOLUTION [0009] According to an exemplary embodiment of the present invention, there is provided a composition for preparing psicose comprising: a microorganism of the genus Kaistia. 45 [0010] The microorganism of the genus Kaistia of the present invention may comprise any microorganism without limitation as long as it has an ability to convert D-fructose to D-psicose. In an exemplary embodiment of the present invention, the microorganism of the genus Kaistia of the present invention may be at least one microorganism of the genus Kaistia selected from the group consisting of Kaistia granuli, Kaistia defluvii, Kaistia adipata, Kaistia geumhonensis, Kaistia dalseonensis, Kaistia hirudinis, Kaistia soli and Kaistia terrae. Specifically, the microorganism of the genus Kaistia 50 of the present invention may be at least one microorganism of the genus Kaistia selected from the group consisting of Kaistia granuli LIS1 (accession number KCCM11916P), Kaistia defluvii LIS2 (accession number KCCM12020P), Kaistia granuli KCTC12575, Kaistia defluvii KCTC23766, Kaistia adipata KCTC12095, Kaistia geumhonensis KCTC 12849, Kaistia dalseonensis KCTC 12850, Kaistia hirudinis DSM25966, Kaistia soli DSM 19436 and Kaistia terrae DSM21341. [0011] In an exemplary embodiment, the microorganism of the genus Kaistia of the present invention may be a strain 55 itself, a culture thereof, or a disruption of the microorganism. The culture or disruption of the microorganism of the genus Kaistia of the present invention may comprise a D-psicose-3-epimerase prepared from the microorganism of the genus Kaistia. In addition, the culture of the microorganism of the genus Kaistia of the present invention may or may not include the microorganism. Further, the disruption of the microorganism of the genus Kaistia of the present invention may be a 2 EP 3 543 345 A2 disruption obtained by disrupting a microorganism of the genus Kaistia or a culture thereof, or a supernatant obtained by centrifuging the disruption. [0012] In another exemplary embodiment of the present invention, the composition for preparing psicose of the present invention may further comprise: D-fructose. 5 [0013] In another exemplary embodiment of the present invention, the microorganism of the genusKaistia of the present invention may be immobilized on a carrier to be used. An example of the carrier that is able to be used in the present invention includes, but is not limited to, agar, agarose, k-carrageenan, alginate or chitosan. [0014] The composition for preparing the D-psicose of the present invention may further comprise a metal. More specifically, the metal of the present invention may be at least one metal selected from the group consisting of manganese, 10 calcium, magnesium, iron, lithium and sodium. More specifically, the metal may be a metal ion or a metal salt, and more specifically, the metal salt may be at least one metal salt selected from the group consisting of LiCl, Na2SO4, MgCl2, NaCl, FeSO4, MgSO4, MnCl2, MnSO4, and CaCl2. The metal ion or the metal salt of the present invention may have a concentration of 0.1 mM to 10 mM, 0.1 mM to 7 mM, 0.1 mM to 4 mM, 0.5 mM to 10 mM, 0.5 mM to 7 mM, 0.5 mM to 4 mM, 1 mM to 10 mM, 1 mM to 7 mM, 1 mM to 4 mM, 2 mM to 10 mM, 2 mM to 7 mM or 2 mM to 4 mM. 15 [0015] According to another exemplary embodiment of the present invention, there is provided a method for preparing psicose, comprising: contacting a microorganism of the genus Kaistia or the composition for preparing psicose described in the present invention with D-fructose. [0016] In an exemplary embodiment, the contacting of the present invention may be performed at pH of 5.0 to 9.0, at 40 to 90°C, and/or for 0.5 to 48 hours. 20 [0017] Specifically, the contacting of the present invention may be performed at pH 6.0 to 9.0, pH 7.0 to 9.0, pH 7.5 to 9.0, pH 6.0 to 8.5, pH 7.0 to 8.5, or pH 7.5 to 8.5. [0018] In addition, the contacting of the present invention may also be performed at a temperature of 40°C to 80°C, 40°C to 75°C, 40°C to 65°C, 50°C to 90°C, 50°C to 80°C, 50°C to 75°C, 50°C to 65°C, 55°C to 90°C, 55°C to 80°C, 55°C to 75°C, 55°C to 65°C, 60°C to 90°C, 60°C to 80°C, 60°C to 75°C, 60°C to 65°C, 65°C to 90°C, 65°C to 80°C or 25 65°C to 75°C. [0019] In addition, the contacting of the present invention may be performed for 0.5 hour or more, 1 hour or more, 3 hours or more, 4 hours or more, 5 hours or more, or 6 hours or more, and/or 48 hours or less, 36 hours or less, 24 hours or less, 18 hours or less, 12 hours or less, 9 hours or less. [0020] In another exemplary embodiment of the present invention, a weight ratio of the microorganism of the genus 30 Kaistia to D-fructose of the present invention may be 1 : 1 to 1 : 5.
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