(19) TZZ ZZ _T (11) EP 2 900 826 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12P 7/18 (2006.01) B01D 11/04 (2006.01) 13.07.2016 Bulletin 2016/28 C07C 29/76 (2006.01) C07C 29/80 (2006.01) (21) Application number: 13785666.2 (86) International application number: PCT/PL2013/000124 (22) Date of filing: 27.09.2013 (87) International publication number: WO 2014/051448 (03.04.2014 Gazette 2014/14) (54) A METHOD FOR ISOLATION OF PROPANE-1,3-DIOL FROM POST-FERMENTATION BROTH OBTAINED BY BIOCONVERSION VERFAHREN ZUR ISOLIERUNG VON PROPAN-1,3-DIOL AUS EINER BRÜHE NACH FERMENTATION HERGESTELLT DURCH BIOCONVERSION UNE MÉTHODE D’ISOLEMENT DE PROPANE-1 ,3-DIOL OBTENU PAR BIOCONVERSION D’UN BOUILLON APRÈS FERMENTATION (84) Designated Contracting States: (56) References cited: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB US-A1- 2012 184 783 US-B2- 7 056 439 GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR • DATABASE WPI Week 200420 Thomson Scientific, London, GB; AN 2004-204156 (30) Priority: 28.09.2012 PL 40097912 XP002719064, & CN 1 460 671 A (UNIV DALIAN POLYTECHNIC) 10 December 2003 (2003-12-10) (43) Date of publication of application: cited in the application 05.08.2015 Bulletin 2015/32 • GUNEET KAUR ET AL: "Advances in biotechnologicalproduction of 1,3-propanediol", (73) Proprietor: Prochimia Surfaces Sp. Z O.O. BIOCHEMICAL ENGINEERING JOURNAL, 81-823 Sopot (PL) ELSEVIER, AMSTERDAM, NL, vol. 64, 7 March 2012 (2012-03-07), pages 106-118, XP028411971, (72) Inventors: ISSN: 1369-703X, DOI: • LINDSTAEDT, Agnieszka 10.1016/J.BEJ.2012.03.002 [retrieved on 84-217 Jelenska Huta (PL) 2012-03-16] • WITT, Dariusz • ZHI-LONG XIU ET AL: "Present state and PL-80-299 Gdansk (PL) perspective of downstream processing of • PUZEWICZ-BARSKA, Joanna biologically produced 1,3-propanediol and PL-81-578 Gdynia (PL) 2,3-butanediol", APPLIED MICROBIOLOGY AND • BARSKI, Piotr BIOTECHNOLOGY, SPRINGER, BERLIN, DE, vol. PL-81-823 Sopot (PL) 78, no. 6, 5 March 2008 (2008-03-05), pages 917-926, XP019586363, ISSN: 1432-0614 (74) Representative: Wazynska, Miroslawa JWP Rzecznicy Patentowi Remarks: Dorota Rzazewska sp.j. Thefile contains technical information submitted after Sienna Center the application was filed and not included in this ul. Zelazna 28/30 specification 00-833 Warszawa (PL) Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 900 826 B1 Printed by Jouve, 75001 PARIS (FR) 1 EP 2 900 826 B1 2 Description isolation from post-fermentation broth containing trimeth- ylene glycol in the range of (30-700g/L) consisting in an [0001] The present invention relates to a method for initial saturation with inorganic salts (10-100%) and or- isolation of propane-1,3-diol from post-fermentation ganic solvent addition for extraction. The method con- broth obtained by bioconversion. 5 cerns specific fermentation broth with trimetylene glycol Propane-1,3-diol is mainly used, in particular as mono- and comprises a technically inconvenient and expensive mer, to build polymer PTT (polytrimethylene terephtha- step of inorganic salt addition. Application of more than late) used in production of fibers, carpets, textiles, for three- carbon ketons (C3) is omitted. CN 1907929 dis- sanitary and medical applications, and as a packaging closes a method of isolation of propane-1,3-diol from fer- and/or structural material. 10 mentation broth containing trimethylene glycol by extrac- As used herein "bioconversion" is a synonym of any fer- tion combined with chemical reaction. The recovery of mentation process leading to enzymatic/biochemical propane-1,3-diol is made after hydrolysis and distillation. way of production of propane-1,3-diol. This is a costly and multistage process which reduces [0002] US 7 056 439 B2 discloses a method for extrac- efficiency thereof and has negative impact on its profit- tion of propane-1,3-diol from fermentation broth with al- 15 ability. cohols (pentanol, propanol, hexanol, oleyl alcohol), ke- Similar recovery technique is presented by Malinowski tones (4-methyl-2-pentanone), esters (isopropyl acetate, et al. (Biotechnol. Prog., 16: 76-79. 2000). This extraction tributyl phosphate), oleic acid, soybean oil, alkanes (n- is based on isolation of propane-1,3-diol by extraction hexane) and mixtures thereof. This method requires mul- with aldehydes, as reactants, converting propane-1,3- tiple extraction with different solvents and reverse extrac- 20 diol into alkyl dioxane and the product is next extracted tion to aqueous phase. Maximum purity of isolated pro- with organic solvents (toluene, o-xylene, ethylbenzene). pane-1,3-diol is 95-98.5%. [0004] Process of propane-1,3-diol extraction from US 5 008 473 describes a method for isolation of pro- post-fermentation broth with mixture of ethyl acetate and pane-1,3-diol with cyclohexane. This method is not suit- ethanol is known in the art from Separation Science and able for mixtures obtained in biotechnological processes 25 Technology, 45: 541-547, 2010. and containing glycerol. [0005] Propane-1,3-diol can also be recovered by con- From EP 1 720 814 B1 a method is known for isolation tinuous liquid-liquid extraction (Biotechnol. Tech., 13: of hydrophilic organic compounds, including propane- 127-130, 1999) however, this process is reported to be 1.3-diol, with glycol ether at a temperature and then as of limited efficiency due to a low distribution ratio of pro- a result of heating to a higher temperature to obtain an 30 pane-1,3-diol in organic solvents. aqueous extract with hydrophilic compound and ether [0006] CN1460671A discloses a step of thermal con- rafinate. The method is based on extraction with ether centration for reduction of volume of the post-fermenta- glycols and a phenomenon specific for them of decreas- tion broth (containing the product) and biomass removal ing the ether solubility in aqueous phase with the increase step by precipitation with organic solvent. Liquid-liquid of temperature. The method discloses also addition of 35 extraction and aqueous/organic phase mixing are not organic solvents in order to further decrease the glycol mentioned there. Mixing is performed only during bio- ethers solubility in aqueous phase during reverse extrac- mass washing after precipitation. Methyl-ethyl ketone is tion step to aqueous phase. Among additives some ke- added, as a precipitating agent, only to remove the bio- tones are here mentioned In this method organic solvent mass, but not as the extractant. This is evidenced by was not used alone for extraction per se. This function 40 absence of phase distribution (aqueous/organic) and ex- was fulfilled by glycol ether, that by nature, forces the traction of the supernatant was not performed. temperature increase and reverse extraction to aqueous [0007] The solution of CN1460671A describes a proc- phase. The whole solution thus constitutes the concen- ess for distillation of mixture of water (from post-fermen- tration of propane-1,3-diol in aqueous phase, as in US 7 tation broth) and an organic solvent (used for precipita- 056 439 B2. 45 tion of biomass). The solution described lacks in the crit- [0003] US 20120184783 discloses a process for puri- ical liquid-liquid extraction process. fication of particular diols, including propane-1,3-diol, [0008] In the solution of the invention the method has from diols mixture by extraction with an organic solvent no concentration or precipitation steps and organic sol- or a mixture of organic solvents. A proper selection of vent is used for liquid-liquid extraction and not for bio- solvent mixture allows to recover a selected diol group 50 mass precipitation. (comprising three, four and five carbon atoms), from the [0009] In the present invention, contrary to the solution starting mixture, where content of water is max. 50 wt.%. of CN1460671A, the extracted substance (propane-1,3- Presented method is not effective however to separate diol) in the liquid-liquid extraction becomes distributed diols from post-fermentation broth with glycerol content between a primary solution (post-fermentation broth) and of less than 5 wt.% nor selective isolation of propane- 55 a secondary solution (organic solvent - methyl-ethyl ke- 1,3-diol. The method fails to complete the use of four- tone, with which the key compound is extracted). As a carbon ketons C4 (2-butanone). result, a rafinate is obtained (a solution basically without CN101012151A describes a method of propane-1,3-diol propane-1,3-diol) and an extract, containing propane- 2 3 EP 2 900 826 B1 4 1,3-diol. Solely the extract (not all the aqueous phase - that is present in the organic solvent mixture. The method supernatant from steps 2 and 3 as in CN1460671A) is is carried out preferably continuously or stepwise and the further purified (eg. subjected to distillation) to acquire organic solvent used in the extraction method is used pure final product and solvent (extractant). repeatedly. The content of 2-butanone in organic sol- [0010] According to the invention, at the step of solvent 5 vents mixture is minimum 10%, and the volume ratio of recovery and final product purification, due to efficient post-fermentation broth, containing propane-1,3-diol, to counter-current extraction the water content is minimized the mixture of organic solvents is minimum 1:3.