Patent ID: 12227467

DETAILED DESCRIPTION OF THE DISCLOSURE

In this context, a fermentation broth (1) comprising bio-1,3-butanediol and water, said fermentation broth preferably being derived from fermentation of sugars obtained from biomass, is sent to the microfiltration step, obtaining a permeate (2) comprising bio-1,3-butanediol, water and residual impurities (for example, sugars, other organic impurities, salts) which is sent to the nanofiltration step, as well as a retentate (not shown inFIG.1) comprising the cell biomass and any cell debris which may be dried and disposed of in landfill or incinerated, or else may be sent directly to a water treatment plant. At the end of said nanofiltration step, a permeate (3) is obtained comprising bio-1,3-butanediol, water and residual impurities (for example, salts) which is sent to the step of treatment with ion-exchange resins, as well as a retentate (not shown inFIG.1) comprising compounds having high steric hindrance (for example, proteins, divalent salts) which may be sent directly to a water treatment plant. Said permeate (3) is thus subjected to the step of treatment with ion-exchange resins, namely to a treatment in a column containing a weak anionic resin followed by a treatment in a column containing a strong cationic resin, or vice versa (not shown inFIG.1), obtaining an aqueous solution (4) comprising bio-1,3-butanediol, water, light organic compounds (for example, ethanol), heavy organic compounds [for example, 4-hydroxy-2-hutanone (4-OH-2B)], unconverted sugars and any unremoved salts. Said aqueous solution (4) is sent to the first evaporation step, obtaining a concentrated solution (5) comprising bio-1,3-butanediol, water, heavy organic compounds [for example, 4-hydroxy-2-butanone (4-OH-2B)], unconverted sugars and any unremoved salts which is sent to the second evaporation step, as well as an aqueous phase (not shown inFIG.1) comprising primarily water and traces of ethanol and of bio-1,3-butanediol which may be recycled to one of the following steps: (a1a) microfiltration (4c), (a2) nanofiltration (4b), (b) treatment with ion-exchange resins. (4a) (shown in dashed lines inFIG.1). From said second evaporation step, a concentrated solution (6) is obtained comprising bio-1,3-butanediol, water and traces of heavy organic compounds [for example, 4-hydroxy-2-butanone (4-OH-2B)], of unconverted sugars and of any unremoved salts which is sent to the third evaporation step, as well as an aqueous phase (not shown inFIG.1) comprising primarily water, organic compounds having a boiling point lower than that of bio-1,3-butanediol and traces of bio-1,3-butanediol which may be sent directly to a water treatment plant. From said third evaporation step, two phases are obtained: an aqueous phase (7) comprising purified bio-1,3-butanediol as well as an organic phase (not shown inFIG.1) comprising any organic compounds having a boiling point higher than that of bio-1,3-butanediol and traces of unpurified bio-1,3-butanediol, of unconverted sugars and of any unremoved salts.

InFIG.2, the mixture (i) containing bio-1,3-butanediol obtained in accordance with the process object of the present invention is fed to a first reactor containing at least one dehydration catalyst, obtaining a stream (ii) comprising alkenols, water, and optionally impurities and/or unreacted bio-1,3-butanediol exiting from said first reactor; said stream (ii) is fed to a first purification section, obtaining a stream (iii) comprising alkenols, water, and, optionally, impurities, a stream (iv) comprising water and, optionally, impurities and/or unreacted bio-1,3-butanediol (not shown inFIG.2), and a stream (v) comprising impurities (not shown inFIG.2); said stream (iii) is fed to a second reactor containing at least one dehydration catalyst, obtaining a stream (vi) comprising bio-1,3-butadiene, water and, optionally, impurities and/or unreacted alkenols exiting from said second reactor, which is fed to a second purification section, obtaining a stream (vii) comprising pure bio-1,3-butadiene, a stream (viii) comprising water and, optionally, unreacted alkenols (not shown inFIG.2) and, optionally, a stream (ix) comprising impurities (not shown inFIG.2).

For better understanding of the present invention and for putting it into practice, illustrative, non-limiting examples thereof are reported hereinafter.

Example 1

Purification of Bio-1,3-Butanediol

For this purpose, a model fermentation broth was used containing bio-1,3-butanediol (referred to hereinafter as bio-1,3-BDO for simplicity) having the average composition reported in Table 1.

TABLE 1Composition of model fermentation broth containing bio-1,3-BDOCompounds*AmountBio-1,3-BDO(g/l)70-95Cell biomass(g/l)20By-products (ethanol, acetic acid, 4-(g/l)20-35hydroxy-2-butanone, proteins)Residual salts(g/l)<10Residual glucose(g/l)<1*balance made up by water.

The content of sugars and organic acids, as well as bio-1,3-butanediol, in diluted solutions (i.e. in aqueous solutions having a concentration of 1,3-BDO<150 g/l) was determined by high-performance liquid chromatography (HPLC), using a Waters 2690 Alliance “module system” chromatograph equipped with a binary pump, degasser, auto-sampler and compartment for the thermostated column. The column used is Phenomenex Rezex ROA-Organic Acid H+, having dimensions 300×7.8 mm, at 45° C., in isocratic with aqueous solution 0.005 N sulphuric acid (H2SO4) at 0.6 ml/min flow rate. Two detectors were used: a Waters 2487 UV/Vis dual absorbance detector (DAD) and a Waters 2410 refractive index detector (RID).

The fermentation broth reported in Table 1, was divided into three separate batches, i.e. “batch 03”, “batch 04” and “batch 05”, and was purified operating as follows.

Removal of Cell Biomass by Microfiltration

To remove the cell biomass, the fermentation broth, i.e. “batch 04” and “batch 05”, was sent to a microfiltration module comprising a Membralox® ceramic membrane in a cross flow configuration having a mean pore diameter of 0.05 μm. Said microfiltration module was subdivided into 19 channels having a diameter of 6 mm and a total filtering area of 0.36 m2. During the test, it was attempted to maintain a flow speed of 4-5 m/s within the membrane, so as to limit the fouling thereof and reach a concentration factor [volume concentration ratio (VCR)], defined as the ratio between the volume of fermentation broth fed in and the volume of permeate obtained, higher than 5, preferably ranging from 6 to 10: further data relating to the operating conditions under which the microfiltration was carried out are reported in Table 2. At the end of the microfiltration, a permeate was obtained comprising bio-1,3-BDO, water and residual impurities (for example, sugars, other organic impurities, salts) which was sent to a nanofiltration module operating as described below, as well as a retentate comprising cell biomass and any cell debris which was dried and disposed of in landfill: the amount of bio-1,3-BDO recovered is also reported in Table 2.

Removal of Cell Biomass by Centrifugation

To remove the cell biomass, the fermentation broth, i.e. “batch 03”, was subjected to centrifugation using a disc centrifuge with automatic panel ejection, model CA 21-P (Andritz), operating under the following conditions:feed rate: 100 litres/h;panel ejection interval: 5 minutes;ejection method: water injection;rotational speed: 8000 rpm;temperature: room temperature (25° C.).

Further data relating to the operating conditions under which the centrifugation was carried out are reported in Table 2. At the end of the centrifugation, a first lighter aqueous phase (supernatant) was obtained comprising bio-1,3-BDO, water and residual impurities (for example, sugars, other organic impurities, salts) which was sent to a nanofiltration module operating as described below, as well as a second heavier aqueous phase (precipitate) comprising the cell biomass and any cell “debris” which was dried and disposed of in landfill: the amount of bio-1,3-BDO recovered is also reported in Table 2.

TABLE 2Operating conditions for the microfiltration (“batch 04” and“batch 05”) and centrifugation (“batch 03”)“batch 03”“batch 04”“batch 05”Centri-Micro-Micro-fugationfiltrationfiltrationFermentation broth(kg)123138.290fed inAverage permeate(kg/m2/h)100(1)72.3107.2flow rateAverage permeate(° C.)≈40(2)52.857.2temperatureFinal VCR—6.29.57.6Transmembrane(bar)—33pressureBio-1,3-BDO end(%)989898balance(3)Bio-1,3-BDO(kg)6.312.45.5recovered(1)flow rate of the feed pump;(2)non-thermostated system;(3)amount of bio-1,3-BDO exiting from the centrifuge or from the microfiltration module divided by the amount of bio-1,3-BDO entering the centrifuge or the microfiltration module × 100.

Nanofiltration

To remove water and residual impurities (for example, sugars, other organic impurities, salts), the nanofiltration step was carried out.

The permeate exiting the microfiltration module, derived from “batch 04” and “batch 05”, was fed to a nanofiltration module containing a spiral-wound membrane, while the first aqueous phase (supernatant) exiting the centrifuge, derived from “batch 03”, was fed to a vibratory nanofiltration system, referred to as a VSEP (vibratory shear-enhanced processing unit—New Logic Research Inc.). Both systems were provided with GE DK membranes (Koch Membrane Systems) having a molecular weight cut-off “(MWCO) ranging from 150 daltons to 300 daltons.

Further data relating to the operating conditions under which the nanofiltration was carried out are reported in Table 3. At the end of the nanofiltration, a permeate was obtained comprising bio-1,3-BDO, water and residual impurities (for example salts, acids) which was sent to the treatment with ion-exchange resins, as well as a retentate comprising compounds having high steric hindrance (for example, proteins, divalent salts) which was dried and disposed of in landfill: the amount of bio-1,3-BDO recovered is also reported in Table 3.

TABLE 3Operating conditions for the nanofiltration (“batch 03”, “batch04” and “batch 05”)“batch 03”“batch 04”“batch 05”VSEPSpiralledSpiralledPermeate fed in(kg)—200166First aqueous phase(kg)121——fed inInitial conductivity(mS/cm)15811Final VCR6.86.29.6Average permeate(kg/h/m2)1643.330.4flow rateAverage temperature(° C.)494142Final conductivity(mS/cm)1344Transmembrane(bar)30.72625.5pressureBio-1,3-BDO end(%)96101101balance(1)Bio-1,3-BDO(kg)5.611.45.4recovered(1)amount of bio-1,3-BDO exiting from the nanofiltration module divided by the amount of bio-1,3-BDO entering the nanofiltration module × 100.

Treatment with Ion-Exchange Resins

To remove residual impurities (for example, salts, acids), the step of treatment with ion-exchange resins was carried out.

For this purpose, the permeate exiting from the nanofiltration module, derived from “batch 03”, was divided into two aliquots, referred to as “batch 03-1” and “batch 03-2” before being sent to the step of treatment with ion-exchange resins.

The permeate exiting the nanofiltration module, derived from “batch 03” (i.e. “batch 03-1” and “batch 03-2”), from “batch 04” and from “batch-05”, was fed to a system from treatment with ion-exchange resins.

The system was composed of two transparent polyvinylchloride columns (PVC-U—GF Piping System) having the following dimensions: diameter=151 mm, height=1200 mm. Said two columns were connected in series, filled with ion-exchange resins: the first column was filled with a strong cationic resin (Dowex™ Monosphere™ 88—Dow Chemical), while the second column was filled with a weak anionic resin (Dowex™ Monosphere™ 77—Dow Chemical).

The purpose of the operation was to obtain a conductivity of the product exiting from the second column<15 μS/cm, and if the exiting product had a higher conductivity said product was fed to said system again after regeneration of the resin.

Said step of treatment with ion-exchange resins was carried out at room temperature (25° C.), while the flow rate was 3.3 BV/h using a metering pump (solenoid-diaphragm metering pump Delta® 4—ProMinent).

Further data relating to the operating conditions under which the step of treatment with ion-exchange resins was carried out are reported in Table 4. At the end of said treatment with ion-exchange resins, an aqueous solution was obtained comprising bio-1,3-BDO, water, light organic compounds (for example, ethanol), heavy organic compounds [for example, 4-hydroxy-2-butanone (4-OH-2B)], unconverted sugars and any unremoved salts which was fed to a rotary evaporator (first evaporation step): the amount of bio-1,3-BDO recovered is also reported in Table 4.

TABLE 4Operating conditions for treatment with ion-exchangeresins (“batch 03-1”, “batch 03-2”, “batch04” and “batch 05”)“batch“batch“batch“batch03-1”03-2”04”05”Permeate fed in(kg)6160240162“Bed Volume”(L resin/6121212column)Permeate conductivity(mS/cm)131344at the enteringProduct conductivity(μS/cm)1.81.60.611.1at the exitingFlow rate(BV/h)3.33.33.33.3Bio-1,3-BDO end(%)100(2)9894balance(1)Bio-1,3-BDO(kg)2.92.611.24.9recovered(1)amount of bio-1,3-BDO exiting from the system for treatment with ion-exchange resins divided by the amount of bio-1,3-BDO entering the system for treatment with ion-exchange resins × 100;(2)total of the amounts obtained from the treatment of “batch 03-1” and “batch 03-2” with ion-exchange resins.

First Evaporation

To eliminate the water and the light organic compounds (for example, ethanol), the aqueous solutions obtained from the step of treatment with ion-exchange resins were subjected to the first evaporation step.

The solutions obtained from the step of treatment with ion-exchange resins were thus fed to a Buchi Rotavapor® rotary evaporator, having a 20-litre loading ball flask, which was heated by immersion in a thermostated water bath. The solution was fed semi-continuously to the loading ball flask and the vapour phase was removed from the evaporator, obtaining a concentrated solution comprising bio-1,3-BDO, water, heavy organic compounds [for example, 4-hydroxy-2-butanone (4-OH-2B)], unconverted sugars and any unremoved salts which was sent to the second evaporation step, as well as an aqueous phase comprising primarily water and traces of ethanol and of bio-1,3-BDO which could be recycled to one of the following steps: (a1a) microfiltration, (a1b) centrifugation, (a2) nanofiltration, (b) treatment with ion-exchange resins.

The conditions for said first evaporation step were as follows:thermostated bath temperature: 60° C.;pressure: 70 mbar-10 mbar.

The evaporation was considered complete at the end of the condensation of the vapours.

The concentrated solutions comprising bio-1,3-BDO were further purified as described in the examples below reported. In particular:the concentrated solution originating from “batch 04” was divided into two aliquots, i.e. “batch 04-1” and “batch 04-2”;the concentrated solutions derived from “batch 03-2” and “batch 04-2” were further purified as described in Example 2 (invention) and in Example 5 (invention) respectively;the concentrated solution derived from “batch 03-1” was further purified as described in Example 3 (comparative);the concentrated solutions derived from “batch 05” and “batch 04-1” were further purified as described in Example 4 (comparative) and in Example 6 (comparative) respectively.

Example 2

Purification of Bio-1,3-BDO (Second and Third Evaporation) (Invention)

The second evaporation and third evaporation steps were carried out as reported below, under the operating conditions reported in Table 5.

For this purpose, the concentrated solution originating from the first evaporation step, i.e. “batch 03-2” (about 2.6 kg), was fed to a rotary evaporator (Laborota® 4003 control—Heidolph) with a 2-litre loading ball flask.

The heating was provided by immersing the loading ball flask in an oil bath equipped with a thermostat. The condenser was formed by a glass coil in the interior of which water and ethylene glycol (about 10%) circulated in a closed circuit. The cooling of the refrigerant fluid was provided by an air/liquid cryostat.

The test started under the operating conditions of the first evaporation step (i.e. 60° C. in the thermostated bath and pressure 70-30 mbar) until absence of evaporation from the sample was verified. Any aqueous phase recovered was discarded. Subsequently, the operating conditions adopted for the second evaporation step and for the third evaporation step are those reported in Table 5 (the evaporation was considered complete at the end of the condensation of the vapours): second evaporation (Step II) and third evaporation (Step III). The amounts of Phase (II) and Phase (III) recovered, as defined below, are also reported in Table 5.

TABLE 5Operating conditions and phases recovered from “batch 03-2”IInd evaporation and IIIrd evaporationTemperature(max)TemperatureEvaporationWeightPressureof oil bathof condenserratePhases(g)(mbar)(° C.)(° C.)(g/h)Phase12812-10110136II(Step II)(Step II)(Step II)(Step II)Phase228912-1012515264III(Step III)(Step III)(Step III)(Step III)

From Step II (second evaporation step) an aqueous phase was obtained comprising water, organic compounds having a boiling point lower than that of bio-1,3-BDO and traces of unpurified bio-1,3-BDO which was disposed of in landfill, as well as a concentrated solution (Phase II) comprising bio-1,3-BDO, water and traces of heavy organic compounds [for example, 4-hydroxy-2-butanone (4-OH-2B)], of unconverted sugars and of any unremoved salts which formed the feed to the third evaporation step (Step III).

From Step III (third evaporation step) an aqueous phase (Phase III) was obtained comprising purified bio-1,3-BDO, as well as an organic phase (Phase IV) comprising any organic compounds having a boiling point higher than that of bio-1,3-BDO and traces of unpurified bio-1,3-BDO, of unconverted sugars and of any unremoved salts.

Table 6 reports the gas chromatography (GC) analysis, carried out as described above in Example 1, of the recovered phases and of the feed (“batch 03-2”).

TABLE 6Composition of “batch 03-2” and of the recovered phases.“batchPhasePhasePhase03-2”IIIIIIVCompo-H2O8.8%1.5%43.0%n.d.sition(3)Bio-1,3-BDO88.0%96.6%34.9%408g/l(%)Bio-4-OH-2B(1)1.0%0.9%5.9%n.d.Decomposition0%0%0%n.d.products of 1,3-BDO(2)Glucose0.1 g/l0 g/l0 g/l15.5g/l(1)bio-4-hydroxy-2-butanone;(2)total of 3-buten-2-one, 2-buten-1-ol (cis and trans isomers), 3-buten-1-ol, 3-buten-2-ol;(3)made up to 100 by other organic impurities not reported in Table 6.

Phase IV had a caramel appearances and was analysed only by high-performance liquid chromatography (HPLC), operating as described above in Example 1.

Meanwhile, for the analysis of Phase II and Phase III, an Agilent HP6890 gas chromatograph (GC) was used, equipped with a Split/Splitless injector on a Quadrex 007 FFAP column of 25 m length, 0.32 mm diameter, 1 μm film, the carrier used was helium at a speed of 50 cm/s, the detector was a flame detector. The determination was carried out using an internal standard with calibration curves for the known individual components. Karl Fischer titration (831 KF Coulometer Metrohm) was further used for the analysis of the water for Phase (III) (for Phase II, the amount of water was calculated by difference).

Phase III was subsequently fed to a dehydration system for the production of butenols, operating as described in Example 7 below reported.

Example 3

Purification of Bio-1,3-BDO (Comparative)

The concentrated solution originating from “batch 03-1”, about 2.7 kg, was fed to a distillation column and processed in batches. The distillation was carried out using a 5-litre boiler and an adiabatic column (diameter=2 cm; height=60 cm) with Sulzer filling. During the distillation, the residual water and the organic compounds having a boiling point between that of water and that of bio-1,3-BDO were removed, whilst the high-boiling organic compounds, the residual salts and the unconverted sugar remained in the boiler: the fractions obtained from the distillation are reported in Table 7.

TABLE 7Fractions distilled from batch 03-1Distillation of batch 03-1TemperatureTemperatureTemperature(max) of(max) ofofDistillationWeightPressureRefluxvapoursboilercondenserrateFraction(g)(mbar)ratio(° C.)(° C.)(° C.)(ml/h)19367536134−51521650569127−5332850588127−55414505116128−54511505121128−52613505124128−5372159502125131−5668183502125140−573

The fractions rich in bio-1,3-BDO are Fraction 7 and Fraction 8. However, all the fractions recovered ended up polluted by a molecule not present in the starting solution; said molecule was identified, using various analytic methods (FT-IR, GC-MS, GC, NMR), as the ketal which forms by reaction between bio-1,3-BDO and 3-buten-2-one (MVK), namely 2,4-dimethyl-2-vinyl-1,3-dioxane (DMV13Diox). In particular, the chromatogram obtained by gas chromatography analysis (GC), carried out as described above in Example 1, made it possible to observe the presence of a new peak in all the fractions, not present in the starting feed. The intensity of the peak in the various fractions ended up different, so different analytical techniques were used for the different fractions.

NMR analysis in solution, FT-IR and GC-MS identified said molecule as 2,4-dimethyl-2-vinyl-1,3-dioxane (DMV13Diox). Said analyses were carried out as follows.

Spectroscopic NMR Analysis of a Sample of Fraction 1

The NMR spectra were recorded using a BrukerAvance 400 NMR tool in deuterated acetone solution. The signal of the CH3of the acetone, positioned at 2.05 ppm in the1H-NMR spectrum and at 29.5 ppm in the13C-NMR spectrum, was taken as a reference signal.

Various products were identified in the1H-NMR spectrum (at different intensities: of these, the most abundant is characterised by the presence of a clearly recognisable vinyl group CH2=CH— at 5.80, 5.32 and 5.29 ppm, corresponding in intensity to a set of signals attributable to CH2—O— and CH—O, characterised, curiously, by very high coupling constants, this generally only being found in cyclic aliphatic systems). Said results made it possible to identify a ketal species, in particular, knowing the primary compounds of the fraction, of the DMV13Diox. All of the couplings were further appreciated only with the aid of a two-dimensional COSY1H-1H spectrum.

Mass Spectrum of a Sample of Fraction 7

Fraction 7 was analysed by gas chromatography-mass spectrometry (GC-MS) on a single-quadrupole spectrometer (Trace DSQ, Thermo), using the headspace method. The sample was measured in a headspace vial and was heated to 80° C. for about one hour. Subsequently, 1 ml of the gaseous phase supernatant on the liquid (area and vapours derived from the sample under examination) and was injected into the gas chromatograph by a method which makes it possible to separate the air (in excess) from the organic components. The analysis conditions used were as follows: temperature scheme of the gas chromatograph from 50° C. to 300° C. at 4° C./minute, injection in split mode, temperature of injector 280° C., transfer line 250° C. Mass spectra recorded from 35 daltons to 500 daltons in electronic ionisation (EI) mode.

Mass Spectrum (EI) for the Species at Retention Time 3.9 Minutes

Although the molecular ion is not visible, it may be assumed that there is a loss of methyl (CH3) from the molecular ion to generate ion 127 (rel. abundance 22.5%) and a loss of 27 (allyl) for ion 115 (rel. abundance 80.9%). A molecular weight of 142 is thus assumed, which would confirm the hypothesis put forward on the basis of the NMR analysis reported above. Starting from the structure hypothesised from the NMR analysis, it would also be possible to explain the other fragment ions obtained in the EI-MS spectrum (indeed, the EI technique generally includes high fragmentation of the organic molecules, and thus makes it possible to reconstruct or confirm the structure). In particular:the main ion 55 (rel. abundance 100%) could correspond to the species CH2═CHCHCH3+*,ion 71 (rel. abundance 35.3%) species CH2═CHCHOCH3+*.

Mass Spectrum (EI) for the Species at Retention Time 4.44 Minutes

Main ion 55 (rel. abundance 100%) species CH2═CHCHCH3+*, ion 71 (rel. abundance 19.3%) species CH2═CHCHOCH3+*.

Ion 127 (rel. abundance 32.5%) through loss of methyl from the molecular ion; loss of 27 (allyl) from the molecular ion for ion 115 (rel. abundance 11.9%).

FT-IR Spectrum of a Sample of Fraction 1

The FT-IR spectra of the samples, deposited on a KBr window, were acquired using a Nicolet Nexus FT-IR spectrometer with 64 scans and a resolution of 2 cm−1.

The bands attributable to the vinyl groups of the product DMV13Diox were found at 3088 cm−1(CH2asymmetric stretching) at 985 cm−1and 916 cm−1(CH and CH2wagging), while the bands in the range 1200 cm−1-1100 cm−1and at 962 cm−1are attributable to the asymmetrical and symmetrical stretching respectively of the C—O bond. Likewise from the product DMV13Diox, the vibrations of the C—O—C bond at 1050 cm−1and of the C═C double bond at 1640 cm−1could be identified.

Table 8 reports the gas chromatography (GC) analysis, carried out as described in Example 1, of the feed (“batch 03-1”), of Fraction 7 and Fraction 8 derived from the distillation of “batch 03-1” and of the boiler residue obtained at the end of the distillation.

TABLE 8GC analysis of the fractions rich in bio-1,3-BDO“batchFrac-Frac-Boiler03-1”tion 7tion 8residueComposition(3)H2O3.1%0.1%0.1%n.d.(%)Bio-1,3-BDO88.3%98.0%97.5%544g/lBio-4-OH-2B(1)1.6%0%0%n.d.DMV13Diox(2)0%0.4%0.2%n.d.Glucosen.d.0 g/l0 g/l5.3g/l(1)bio-4-hydroxy-2-butanone;(2)2,4-dimethyl-2-vinyl-1,3-dioxane (expressed as percentage area based on the area of the peak for the bio-1,3-BDO);(3)made up to 100 by other organic impurities not reported in Table 8.

The boiler residue had a caramel appearance and was analysed only by HPLC, operating as described in Example 1 above. Karl Fischer titration (831 KF Coulometer Metrohm) was further used for the analysis of the water for Fraction 7 and for Fraction 8.

Example 4

Purification of Bio-1,3-BDO (Comparative)

The solution originating from “batch 05”, about 4.2 kg, was fed to a distillation column and processed in batches. The distillation was carried out using a 5-litre boiler and an adiabatic column (diameter=2 cm; height=60 cm) with Sulzer filling. During the distillation, the residual water and the organic compounds having a boiling point between that of water and that of bio-1,3-BDO were removed, while the high-boiling organic compounds, the residual salts and the unconverted sugar remained in the boiler; Table 9 reports the fractions obtained from the distillation.

TABLE 9Fractions distilled from “batch 05”Distillation of “batch 05”VapourBoilertemperaturetemperatureCondenserDistillationWeightPressureReflux(max)(max)temperaturerateFraction(g)(mbar)ratio(° C.)(° C.)(° C.)(ml/h)1956353229516215810570105127314210157494194122101577951951111015739619632771238510710327135123851371034

The fractions rich in 1,3-BDO are Fraction 6 and Fraction 7.

By reducing the pressure in the column, and thus the temperature of the system, formation of 3-buten-2-one (MVK) and of the corresponding ketal is avoided. However, in the last fraction distilled (Fraction 7), the presence of decomposition products of bio-1,3-BDO, which lead to a decrease in the yield of the distillation, is noted: Table 10 reports the gas chromatography analysis, carried out as described in Example 1 above, of the fractions rich in bio-1,3-BDO.

TABLE 10GC analysis of the fractions rich in 1,3-BDO “batch 05”.Boiler“batch 05”Fraction 5Fraction 6Fraction 7residueComposition(4)H2O4.3%0.6%0.3%0.9%n.d.(%)Bio-1,3-BDO91.3%96.5%98.2%97.9%266g/lBio-4-OH-2B(1)0.6%0%0%0%n.d.DMV13Diox(2)0%0%0%0%n.d.Decomposition0%0%0.01%0.58%n.d.products of 1,3-BDO(3)Glucose0.2 g/l0 g/l0 g/l0 g/l7.6g/l(1)bio-4-hydroxy-2-butanone;(2)2,4-dimethyl-2-vinyl-1,3-dioxane (expressed as percentage area based on the area of the peak for bio-1,3-BDO);(3)total of buten-2-one, 2-buten-1-ol (cis and trans isomers), 3-buten-1-ol, 3-buten-2-ol;(4)made up to 100 by other organic impurities not reported in Table 10.

The boiler residue had a caramel appearance and was analysed only by HPLC, operating as described in Example 1 above. Karl Fischer titration (831 KF Coulometer Metrohm) was further used for the analysis of the water for Fraction 5, for Fraction 6 and for Fraction 7.

Fraction 6 of “batch 5” was subsequently fed to a dehydration system for the production of butenols, operating as described in Example 7 below.

Example 5

Purification of Bio-1,3-BDO (Second and Third Evaporation) (Invention)

The second and third evaporation steps were carried out as reported below, under the operating conditions reported in Table 11.

For this purpose, the concentrated solution originating from the first evaporation step, i.e. “batch 04-2” (about 6.8 kg), was fed to a rotary evaporator (Laborota® 4003 control—Heidolph) with a 2-litre loading ball flask.

The heating was provided by immersing the loading ball flask in an oil bath equipped with a thermostat. The condenser was formed by a glass coil in the interior of which water and ethylene glycol (about 10%) circulated in a closed circuit. The cooling of the refrigerant fluid was provided by an air/liquid cryostat.

The test started under the operating conditions of the first evaporation step (i.e. 60° C. in the thermostated bath and pressure 70-30 mbar) until absence of evaporation from the sample was verified. Any aqueous phase recovered was discarded. Subsequently, the operating conditions adopted for the second evaporation step and for the third evaporation step are those reported in Table 11 (the evaporation was considered complete at the end of the condensation of the vapours): second evaporation (Step II) and third evaporation (Step III). The amounts of Phase (II) and Phase (III) recovered, as defined below, are also reported in Table 11.

TABLE 11Operating conditions and phases recovered from “batch 04-2”IInd evaporation and IIIrd evaporationTemperature(max)CondenserEvaporationWeightPressureof oil bathtemperatureratePhases(g)(mbar)(° C.)(° C.)(g/h)Phase46412-10110128II(Step II)(Step II)(Step II)(Step II)Phase682712-1012515338III(Step III)(Step III)(Step III)(Step III)

From Step II (second evaporation step) an aqueous phase was obtained comprising water, organic compounds having a boiling point lower than that of bio-1,3-BDO and traces of unpurified bio-1,3-BDO which was disposed of in landfill, as well as a concentrated solution (Phase II) comprising bio-1,3-BDO, water and traces of heavy organic compounds (for example, 4-hydroxy-2-butanone), of unconverted sugars and of any unremoved salts which formed the feed to the third evaporation step (Step III).

From Step III (third evaporation step) an aqueous phase (Phase III) was obtained comprising purified bio-1,3-BDO, as well as an organic phase (Phase IV) comprising any organic compounds having a boiling point higher than that of bio-1,3-BDO and traces of unpurified bio-1,3-BDO, of unconverted sugars and of any unremoved salts.

Table 12 reports the gas chromatography (GC) analysis, carried out as described above in Example 1, of the recovered phases and of the feed (“batch 04-2”).

TABLE 12Composition of “batch 04-2” and of the recovered phases.“batchPhasePhasePhase04-2”IIIIIIVCompo-H2O4.2%0.7%52.8%n.d.sition(3)Bio-1,3-BDO94.1%97.8%34.9%374g/l(%)Bio-4-OH-2B(1)1.1%0.8%5.9%n.d.Decomposition0%0%0%n.d.products of 1,3-BDO(2)Glucose0.4 g/l0 g/l0 g/l26.9g/l(1)bio-4-hydroxy-2-butanone;(2)total of 3-buten-2-one, 2-buten-1-ol (cis and trans isomers), 3-buten-1-ol, 3-buten-2-ol;(3)made up to 100 by other organic impurities not reported in Table 12.

Phase IV had a caramel appearances and was analysed only by high-performance liquid chromatography (HPLC), operating as described above in Example 1.

Karl Fischer titration (831 KF Coulometer Metrohm) was further used for the analysis of the water for Phase (III) (for Phase II, the amount of water was calculated by difference).

Phase III was subsequently fed to a dehydration system for the production of butenols, operating as described in Example 7 below reported.

Example 6

Purification of Bio-1,3-BDO (Comparative)

The solution originating from “batch 04-1”, about 4.05 kg, was fed to a distillation column and processed in batches. The distillation was carried out using a 5-litre boiler and an adiabatic column (diameter=2 cm; height=60 cm) with Sulzer filling. During the distillation, the residual water and the organic compounds having a boiling point between that of water and that of bio-1,3-BDO were removed, while the high-boiling organic compounds, the residual salts and the unconverted sugar remained in the boiler: Table 13 reports the fractions obtained from the distillation.

TABLE 13Fractions distilled from “batch 04-1”Distillation of “batch 04-1”VapourBoilertemperaturetemperatureCondenserDistillationWeightPressureReflux(max)(max)temperaturerateFraction(g)(mbar)ratio(° C.)(° C.)(° C.)(ml/h)11243032497−116270123082102563260123841035554344612386110553526123841181045

The fractions rich in 1,3-BDO are Fraction 4 and Fraction 5.

By reducing the pressure in the column, and thus the temperature of the system, formation of 3-buten-2-one (MVK) and of the corresponding ketal is avoided. However, in the last fraction distilled (Fraction 5), the presence of decomposition products of bio-1,3-BDO, which lead to a decrease in the yield of the distillation, is noted: Table 14 reports the gas chromatography (GC) analysis, carried out as described in Example 1 above reported, of the fractions rich in bio-1,3-BDO and of the feed (“batch 04-1”).

TABLE 14GC analysis of fractions rich in 1,3-BDO “batch 04-1”.boiler“batch 04-1”Fraction 3Fraction 4Fraction 5residueComposition(4)H2O2.6%0.4%0.1%n.d.n.d.(%)Bio-1,3-BDO95.5%97.5%99%99%536g/lBio-4-OH-2B(1)0.9%0.5%0%0%n.d.DMV13Diox(2)0%0%0%0%n.d.Decomposition0%0.04%0%0.03%n.d.products of 1,3-BDO(3)Glucose0.1 g/l0 g/l0 g/l0 g/l1.2g/l(1)bio-4-hydroxy-2-butanone;(2)2,4-dimethyl-2-vinyl-1,3-dioxane (expressed as percentage area based on the area of the peak for bio-1,3-BDO);(3)total of 3-buten-2-one, 2-buten-1-ol (cis and trans isomers), 3-buten-1-ol, 3-buten-2-ol;(4)made up to 100 by other organic impurities not reported in Table 14.

The boiler residue had a caramel appearance and was analysed only by HPLC, operating as described in Example 1 above. Karl Fischer titration (831 KF Coulometer Metrohm) was further used for the analysis of the water for Fractions 3, 4 and 5.

Fraction 4 of “batch 04-1” was subsequently fed to a dehydration system for the production of butenols, operating as described in Example 7 below reported.

Table 15 reports the yields of Example 2 (invention) (Phase III of “batch 03-2”), Example 4 (comparative) (Fraction 6 of “batch 05”), Example 5 (invention) (Phase III of “batch 04-2”) and Example 6 (comparative) (Fraction 4 of “batch 04-1”). The yields of Example 3 (comparative) are not reported, since the fractions rich in 1,3-BDO are polluted by 2,4-dimethyl-2-vinyl-1,3-dioxane (DMV13Diox) and are not sent to a dehydration system for the production of butenols.

TABLE 15Yield and recovery rate of the fractions of Example 1 and Example4 (invention), Example 3 and Example 5 (comparative)GlobalRecoveryBio-1,3-massof fractionTotalBDOGlobalbalance ofrich inrecoveryrecoverymassbio-1,3-bio-1,3-rateratebalanceBDOBDO(g/h)(g/h)Example 2100%99%97%139264(invention)(“batch03-2”)Example 499%99%84%2432(comparative)(“batch05”)Example 5101%99%96%192338(invention)(“batch04-2”)Example 6100%97%88%4453(comparative)(“batch04-1”)

The total recovery rate was calculated as the total of the recovered phases or fractions (g) divided by the overall hours of testing (h), counted from the first condensed drop removed. The bio-1,3-BDO recovery rate was calculated as the amount of purified phase fraction recovered (g) divided by the hours to purify this individual phase or fraction (h), counted from the first condensed drop removed of said phase or fraction.

From the data reported in Table 15, it may be seen that Example 2 and Example 5 (invention) end up having a higher yield (i.e. recovery of phase or fraction rich in bio-1,3-BDO) than Example 4 and Example 6 (comparative), and the bio-1,3-BDO recovery rate from the phase or fraction used ends up much higher.

Example 7

Production of Bio-Butenols from Bio-1,3-BDO

The purified 1,3-BDO obtained in Example 2 (invention) (Phase III of “batch 03-2”) and the purified bio-1,3-BDO obtained in Example 4 (comparative) (Fraction 6 of “batch 05”) were subjected to catalytic dehydration, operating as follows.

For this purpose, the catalysts A and B were first synthesised, operating as follows.

Synthesis of Catalyst A

In a glass beaker, provided with magnetic bar stirrer, was prepared a solution of 870 g of cerium nitrate hexahydrate in 4200 g water, by vigorous stirring, at room temperature (25° C.). The solution obtained was transferred into a glass reactor, equipped with stirring rod in the form of a bar, and was kept under stirring for 15 minutes. To the solution obtained, kept under stirring, were added, by peristaltic pump, 790 g of an aqueous solution at 15% of ammonium hydroxide (NH4OH), prepared previously by diluting the commercial aqueous solution at 28%-30%, over 3 hours, while monitoring the pH by Metrohm glass pH electrode (6.0248.030), connected to the Metrohm 691 pH-meter. At the end of adding the solution, the pH of the suspension was 9.0, the whole was left under stirring in the same conditions for 64 hours, at the end of which the pH ended up at 4.3. Subsequently, to the suspension obtained, kept under stirring, was added, by peristaltic pump, a further 90 g of an aqueous solution at 15% of ammonium hydroxide (NH4OH), prepared previously as described above, over 25 minutes, obtaining a suspension of pH 9.0. The suspension was left, under stirring, for 24 hours, at the end of which the pH was measured again and ended up at 8.8, and a precipitate was obtained. The precipitate obtained was filtered, washed with about 10 litres of water, and subsequently oven-dried at 120° C., for 2 hours. After drying, the solid obtained was calcined for 6 hours, at 600° C.

Synthesis of Catalyst B

To a beaker, equipped with stirring rod with Teflon crescent blade, were added 200 g of an aqueous solution at about 30% of ammonium hydroxide (NH4OH), and an electrode [Metrohm glass pH electrode (6.0248.030), connected to the Metrohm 780 pH-meter] was introduced to measure the pH. In another beaker, equipped with magnetic bar stirrer, was prepared a solution of 200 g of cerium nitrate hexahydrate in 200 g of water: the cerium nitrate was then solubilised by vigorous stirring at room temperature (25° C.). The solution obtained was introduced into a dropper and fed drop by drop, over 6 minutes, into the aforementioned ammonium hydroxide solution contained in the beaker, under constant vigorous stirring. The pH of the suspension obtained was 10.1. The whole was left, under vigorous stirring, for 3 hours, during which 200 ml water was added and the pH, which was 9.6, was measured. The whole was left, under vigorous stirring, for another 1.5 hours, at the end of which another 200 ml water was added and the pH, which was 9.5, was measured again. Said suspension was left, under vigorous stirring, for 64 hours, at the end of which the pH, which was 4.5, was measured again. Subsequently, another 23 g of an aqueous solution at about 30% of ammonium hydroxide (NH4OH) were added, obtaining a pH of 9.0: the whole was left, under stirring, for 6 hours, obtaining a pH of 8.5. Subsequently, 16 g of an aqueous solution at about 30% of ammonium hydroxide (NH4OH) were added, obtaining a pH of 9.0. The whole was left, under vigorous stirring, for 17 hours, at the end of which the pH was 7.9, and a precipitate was obtained. The precipitate obtained was filtered, washed with 2 litres of water, and subsequently oven-dried at 120° C., for 2 hours. After drying, the solid obtained was calcined for 5 hours, at 600° C.

Dehydration Reaction

The dehydration reaction of the bio-1,3-BDO was carried out in an AISI 316L steel fixed-bed continuous tubular reactor, 400 mm long and with an internal diameter of 9.65 mm. The reactor is thermostated to the reaction temperature using an electric heater. For optimum regulation of the temperature within the reactor, along the axis thereof there is a well having external diameter 3 mm which houses the thermocouple for the temperature regulation.

The catalysts used in the tests are sieved and selected in the fraction ranging from 0.5 mm to 1 mm. The catalyst load, equal to 3 g, was inserted into the reactor between two layers of inert material (corundum), the catalytic bed was held in place by a sintered steel septum placed on the base of the reactor.

The feed-in was carried out from the top of the reactor, above the region filled with inert material, which region also acts as an evaporator and allows the reagents to pass into the gaseous state and reach the reaction temperature before coming into contact with the catalyst. The reactor thus has a down-flow setup.

The liquid reagents were fed in using a metering pump of the type used in high-performance liquid chromatography (HPLC). The gases were fed in by thermal mass flow-meter (TMF). Downstream of the reactor, the products obtained were cooled in a heat exchanger, and the condensed liquid was collected in glass vials using a series of time-controlled valves. The collected samples of liquids were analysed by gas chromatography analysis using an Agilent HP6890 gas chromatograph (GC) equipped with a Split/Splitless injector on a Quadrex 007 FFAP column of 25 m length, 0.32 diameter, 1 μm film, the carrier used was helium at a speed of 50 cm/s, the detector was a flame detector. The determination was carried out using an internal standard with calibration curves for the known individual components.

Meanwhile, the incondensable gases were analysed in an online gas chromatograph (GC) and finally sent to a volumetric drum-type gas meter, to measure the volume. The online analysis of the gases was carried out by an Agilent HP7890 gas chromatograph (GC) with HP-AI/S column of 50 m length, 0.53 mm diameter, 15 microns film, the carrier used was helium at a speed of 30 cm/s, the detector was a flame detector. The determination was carried out using an external standard with calibration curves for the known individual components.

The catalytic performances reported in Table 16 and in Table 17 are expressed by calculating the conversion of bio-1,3-BDO (C1,3-BDO) and the butenols selectivities in accordance with the formulae below reported:

C1,3-BDO=(moles1,3-BOD)in-(moles1,3-BOD)out(moles1,3-BOD)in×100Sbutenols=molesbutenols(moles1,3-BOD)in-(moles1,3-BOD)out

wherein:(moles1,3-BDO)in=moles of 1,3-butanediol in input;(moles1,3-BDO)out=moles of 1,3-butanediol in output;molesbutenols=total moles of alkenols [referring to 3-buten-2-ol (methylvinylcarbinol) and 2-buten-1-ol (crotyl alcohol)].

If the total of the selectivities of all the products exceeds 100%, the result of the test is expressed as selectivity for butenols normalised to 100, or:

Sbutenols⁢⁢normalised=SbutenolsΣ⁢⁢SProducts×100

Before the catalytic test is started, the catalyst is treated in situ at 300° C., in a nitrogen flow (N2). To said reactor are subsequently fed 36 g/h of an 83.3% solution of 1,3-butanediol in water at atmospheric pressure (1 bar absolute).

About 3.19 kg of Fraction 6 of “batch 05”, obtained as described in Example 4 (comparative), were diluted with 640 g water. The solution obtained was fed to the dehydration reactor using catalyst A at a temperature of 370° C. without the catalyst exhibiting problems of loss of activity or loss of yield.

Subsequently, in a second test which used catalyst B, 2.135 kg of Phase III of “batch 03-2”, obtained as described in Example 2 (invention) added with 428 g water were fed. The reaction temperature was set to 355° C. In this case too, the catalyst did not exhibit problems of loss of activity or loss of yield.

Table 16 reports the catalytic results obtained in terms of conversion (C %) and selectivity (S %), calculated as described above.

TABLE 16Results of dehydration tests on purified bio-1,3-BDO obtainedin Example 2 (invention) (Phase III of “batch 03-2”) andExample 4 (comparative) (Fraction 6 of “batch 05”)Temper-Conver-Selec-aturesiontivityYieldCatalyst(° C.)(%)(%)(%)Example 4A370° C.959691(comparative)(Fraction 6 of“batch 05”)Example 2B355° C.989189(invention)(Phase III of“batch 03-2”)

Example 8

The purified bio-1,3-BDO obtained in Example 5 (invention) (Phase III of “batch 04-2”) and the purified bio-1,3-BDO obtained in Example 6 (comparative) (Fraction 4 of “batch 04-1”) were subjected to catalytic dehydration, operating as follows.

For this purpose, the catalyst C was first synthesised, operating as follows.

Synthesis of Catalyst C

Preparation of a Catalyst Based on Extruded Cerium Oxide The extruded catalyst was obtained by operating as described in Example 9 of international patent application WO 2015/173780 in the name of the Applicant.

For this purpose, in a glass beaker, equipped with magnetic bar stirrer, was prepared a solution of 870 g cerium nitrate hexahydrate (99% Aldrich, product code 238538; CAS number 10294-41-4) in 4200 g of water, by vigorous stirring, at room temperature (25° C.). The solution obtained was transferred into a glass reactor, equipped with stirring rod in the form of a bar and was kept, under stirring, for 15 minutes. To the solution obtained, kept under stirring, were added, by peristaltic pump, 790 g of an aqueous solution at 15% of ammonium hydroxide (NH4OH), prepared previously by diluting the commercial aqueous solution at 28%-30% (Aldrich 28%-30% NH3Basis ACS reagent; product code 221228; CAS number 1336-21-6), over 3 hours, while monitoring the pH by Metrohm glass pH electrode 6.0248.030, connected to the Metrohm 691 pH-meter. At the end of adding the solution, the pH of the suspension was 9.0: the whole was left, under stirring, for 64 hours, at the end of which the pH ended up at 4.3. Subsequently, to the suspension obtained, kept under stirring, were added, by peristaltic pump, a further 90 g of an aqueous solution at 15% aqueous of ammonium hydroxide (NH4OH), prepared previously as described above, over 25 minutes, obtaining a suspension of pH 9.0. The suspension was left, under stirring, for 24 hours, at the end of which the pH was measured again and ended up at 8.8, and a precipitate was obtained. The precipitate obtained was filtered, washed with about 10 litres of water, and subsequently oven-dried at 120° C., for 2 hours.

After said preparation had been repeated for a number of batches suitable for obtaining sufficient amounts of material, the solids obtained were combined and ground in a mortar: 1905 g of powder thus obtained were subsequently placed in an Erweka planetary mixer with AMD model motor.

The powder was dry mixed for 1 hour and, subsequently, there were added, by dropping, in sequence, 250 g of an aqueous solution at 25% of ammonium hydroxide (NH4OH), prepared previously by diluting the commercial aqueous solution at 28%-30% (Aldrich 28%-30% NH3Basis ACS reagent; product code 221228; CAS number 1336-21-6), over 50 minutes, and 250 ml demineralised water, likewise over 50 minutes, obtaining a paste which was extruded using a Hutt extruder on which were mounted rollers having 2 mm holes. The pellets obtained from the extrusion were left to air-dry for two days.

Subsequently, a sample of the pellets of weight 134 g was oven-dried at 120° C., for 2 hours, and subsequently calcined for 6 hours, at 600° C., obtaining a catalyst based on cerium oxide. The catalyst is then prepared for the catalytic test by comminuting it and sieving it, selecting the fraction ranging from 0.5 mm to 1 mm.

Catalyst C, obtained as described above, was loaded into the reactor and subjected to the preliminary operations as described in Example 7. The test was started with an 83.3% 1,3-BDO solution of non-biological origin (Sigma-Aldrich B84784) in demineralised water, at a temperature of 370° C. and with a flow rate of 36 g/h.

After about 125 hours of operation, there was fed into the dehydration reactor 4.072 kg of an 83.3% solution of bio-1,3-BDO in water, obtained by suitably diluting Fraction 4 of “batch 04-1” obtained as described in Example 6 (comparative). Table 17 reports the catalytic results obtained in terms of conversion (C %) and selectivity (S %), calculated as described above. During the test, the performance remained constant, and the catalyst did not exhibit problems of loss of activity or loss of yield.

The test was continued using the 1,3-BDO starting solution of non-biological origin. In this step, the reaction temperature was set to 375° C. to recover a slight conversion loss. Subsequently, during the same test, there were fed 4.226 kg of the solution obtained by diluting with demineralised water Phase III of “batch 04-2” obtained as described in Example 5 (invention), so as to obtain an 83.3% solution. In this case too, Table 17 reports the catalytic results obtained in terms of conversion (C %) and selectivity (S %), calculated as described above. As in the previous case, the performance remained constant and the catalyst did not exhibit problems of loss of activity or loss of yield.

TABLE 17Results of dehydration tests on purified bio-1,3-BDO obtainedin Example 6 (comparative) (Fraction 4 - “batch 04-1”) andExample 5 (invention) (Phase III of “batch 04-2”)Temper-Conver-Selec-aturesiontivityYieldCatalyst(° C.)(%)(%)(%)Example 6C370° C.949690(comparative)(Fraction 4 of“batch 04-1”)Example 5C375° C.959489(invention)(Phase III of“batch 04-2”)

Meanwhile, Table 18 reports the overall yield of butenols, understood as being produced between the yield of the last purification step and of the dehydration.

TABLE 18Overall yield of butenolsAverage yield of butenolsExample 2Example 5Example 4Example 6(inven-(inven-(compar-(compar-tion)tion)ative)ative)(Phase III(Phase III(Fraction 6(Fraction 4of “batchof “batchof “batchof “batch03-2)04-2”)05”)04-1”)[% mol][% mol]Distillation——8488(comparative)Evaporation9796——(three steps)(invention)Dehydration89899190to butenolsAverage86857679overall yield

From the data reported in Table 18, it may be seen that the process according to the present invention for producing bio-butenols from a fermentation broth is an improvement on the prior art in terms of average overall yield.