Compositions and methods for purification of isoprenoids

Abstract

To provide a method for producing a high-purity isoprenoid from an isoprenoid composition.SOLUTION: The method includes using chromatography to reduce impurities in the isoprenoid composition to below parts per million levels.SELECTED DRAWING: None

Description

[Background technology]

[0001] Isoprenoids are ubiquitous in nature. They comprise a diverse family of over 40,000 individual molecules, many of which are essential for living organisms. Isoprenoids play a role in maintaining cellular fluidity, electron transport, and other metabolic functions. A vast number of natural and synthetic isoprenoids are useful as pharmaceuticals, cosmetics, fragrances, pigments and colorants, fungicides, disinfectants, dietary supplements, and fine chemical intermediates. Given the large quantities of isoprenoid products required for many commercial applications and the utility of isoprenoids as polymers, there remains a need to develop methods for producing isoprenoids.

[0002] Isoprenoids can be produced from host cells, extracted, and then purified. In some cases, isoprenoids can be extracted and purified through processes such as centrifugation, distillation, and the like. Purified isoprenoids can have a purity of 95% (w / w) or more. However, before isoprenoids can be used in certain applications, such as as monomers in polymerization reactions, it may still be necessary to reduce certain impurities, such as oxygenates, to parts per million levels. Therefore, there remains a need to develop methods for purifying isoprenoids to a purity suitable for applications such as polymerization reactions. Summary of the Invention

[0003] The present disclosure provides compositions and methods for isolating highly pure isoprenoids from isoprenoid compositions, as well as compositions of the highly pure isoprenoids. The compositions and methods described herein address a particularly challenging problem: how to obtain isoprenoids from available sources (e.g., distilled isoprenoids) with a purity high enough to be used in commercially useful applications, such as polymerization reactions. Although isoprenoids can be produced using various methods, such as by distilling isoprenoids from host cells expressing enzymes involved in isoprenoid biosynthesis or from other sources, the purity of the resulting isoprenoids may not be high enough to be effectively and consistently used in applications such as polymerization reactions. Therefore, in order to enable the isoprenoids to be used in applications such as polymerization reactions, it is necessary to purify the isoprenoids so that impurities are present at very low levels (e.g., parts per million). Given the extremely high level of purity required, achieving such highly pure isoprenoids presents significant challenges.

[0004] This disclosure is based, in part, on the surprising discovery that the unique combination of chromatography steps and resin treatments described herein results in isoprenoid compositions having extremely high levels of purity that were not achievable by other tested methods. The following sections describe compositions and methods that can be used to obtain highly pure isoprenoids from isoprenoid compositions.

[0005] In an aspect, the disclosure provides a method for purifying farnesene, the method comprising: (a) providing a farnesene composition; and (b) purifying the farnesene from the farnesene composition of (a) by chromatography.

[0006] In some embodiments, the farnesene composition comprises farnesene and one or more impurities. In some embodiments, the one or more impurities comprise one or more polar impurities. In some embodiments, the one or more polar impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0007] In some embodiments, the farnesene composition has a concentration of one or more impurities from 1 ppm to 15,000 ppm (e.g., from 1 ppm to 10,000 ppm, from 1 ppm to 5,000 ppm, from 1 ppm to 1,0000 ppm, from 1 ppm to 500 ppm, from 1 ppm to 100 ppm, from 100 ppm to 15,000 ppm, from 1,000 ppm to 15,000 ppm, from 5,000 ppm to 15,000 ppm, from 10,000 ppm to 15,000 ppm, from 5,000 ppm to 12,000 ppm, from 1,000 ppm to 10,000 ppm, or from 3,000 ppm to 14,000 ppm). In some embodiments, the farnesene composition has a concentration of one or more impurities from 1000 ppm to 12,000 ppm (e.g., from 1000 ppm to 10,000 ppm, from 1000 ppm to 8,000 ppm, from 1000 ppm to 5,000 ppm, from 1000 ppm to 2,000 ppm, from 2,000 ppm to 12,000 ppm, from 5,000 ppm to 12,000 ppm, from 8,000 ppm to 12,000 ppm, from 10,000 ppm to 12,000 ppm, from 5,000 ppm to 10,000 ppm, or from 3,000 ppm to 10,000 ppm). In some embodiments, the farnesene composition has a concentration of one or more impurities from 100 ppm to 1,000 ppm (e.g., from 100 ppm to 800 ppm, from 100 ppm to 600 ppm, from 100 ppm to 400 ppm, from 100 ppm to 200 ppm, from 200 ppm to 1,000 ppm, from 500 ppm to 1,000 ppm, from 800 ppm to 1,000 ppm, or from 300 ppm to 800 ppm).

[0008] In some embodiments, 4-tert-butylcatechol is added to the farnesene composition. In some embodiments, the 4-tert-butylcatechol is added to the farnesene composition to a concentration of 50 ppm to 150 ppm (e.g., 50 ppm to 125 ppm, 50 ppm to 100 ppm, 50 ppm to 75 ppm, 75 ppm to 150 ppm, 100 ppm to 150 ppm, 125 to 150 ppm, or 25 ppm to 75 ppm), optionally, the 4-tert-butylcatechol is added to the farnesene composition to a concentration of about 100 ppm.

[0009] In some embodiments, the chromatography comprises (a) pre-flowing a resin, (b) exposing the farnesene composition to the resin, and (c) recovering the farnesene from the resin. In some embodiments, the resin comprises aluminum oxide. In some embodiments, the aluminum oxide is basic aluminum oxide. In some embodiments, the aluminum oxide is acidic aluminum oxide. In some embodiments, the aluminum oxide is neutral aluminum oxide. In some embodiments, the resin comprises silica.

[0010] In some embodiments, the resin has a bulk density of 0.1 g / mL to 0.75 g / mL (e.g., 0.1 g / mL to 0.5 g / mL, 0.1 g / mL to 0.25 g / mL, 0.2 g / mL to 0.75 g / mL, 0.5 g / mL to 0.75 g / mL, 0.g / mL to 0.75 g / mL, or 0.3 g / mL to 0.5 g / mL). In some embodiments, the resin has a bulk density of 0.25 g / mL to 0.5 g / mL (e.g., 0.25 g / mL to 0.4 g / mL, 0.25 g / mL to 0.3 g / mL, 0.3 g / mL to 0.5 g / mL, 0.4 g / mL to 0.5 g / mL, or 0.3 g / mL to 0.4 g / mL). In some embodiments, the resin has a pore volume of 0.1 mL / g to 1.5 mL / g (e.g., 0.1 mL / g to 1.2 mL / g, 0.1 mL / g to 1 mL / g, 0.1 mL / g to 0.7 mL / g, 0.1 mL / g to 0.5 mL / g, 0.1 mL / g to 0.2 mL / g, 0.2 mL / g to 1.5 mL / g, 0.7 mL / g to 1.5 mL / g, 1 mL / g to 1.5 mL / g, 1.2 mL / g to 1.5 mL / g, or 0.5 mL / g to 1 mL / g). In some embodiments, the resin has a pore volume of 0.5 mL / g to 1 mL / g (e.g., 0.5 mL / g, 0.6 mL / g, 0.7 mL / g, 0.8 mL / g, 0.9 mL / g, or 1 mL / g).

[0011] In some embodiments, the resin has a particle distribution size of 25 μm to 800 μm (e.g., 25 μm to 700 μm, 25 μm to 600 μm, 25 μm to 500 μm, 25 μm to 400 μm, 25 μm to 300 μm, 25 μm to 200 μm, 25 μm to 100 μm, 25 μm to 50 μm, 50 μm to 100 μm, 50 μm to 800 μm, 100 μm to 800 μm, 200 μm to 800 μm, 300 μm to 800 μm, 500 μm to 800 μm, 600 μm to 800 μm, or 50 μm to 200 μm). In some embodiments, the resin has a particle distribution size of 100 μm to 800 μm (e.g., about 100 μm to about 700 μm, about 100 μm to about 600 μm, about 100 μm to about 500 μm, about 100 μm to about 400 μm, about 100 μm to about 300 μm, about 100 μm to about 200 μm, about 200 μm to about 800 μm, about 300 μm to about 800 μm, about 400 μm to about 800 μm, about 500 μm to about 800 μm, about 600 μm to about 800 μm, about 700 μm to about 800 μm, about 200 μm to about 600 μm). In some embodiments, the resin has a particle distribution size of 300 μm to 600 μm (e.g., about 300 μm to about 550 μm, about 300 μm to about 500 μm, about 300 μm to about 450 μm, about 300 μm to about 400 μm, about 300 μm to about 350 μm, about 350 μm to about 600 μm, about 400 μm to about 600 μm, about 450 μm to about 600 μm, about 500 μm to about 600 μm, about 550 μm to about 600 μm, or about 400 μm to about 500 μm).

[0012] In some embodiments, the resin has an approximate moisture content of less than 1% w / w. In some embodiments, the resin has an approximate moisture content of 0.01% w / w to 9% w / w (e.g., 0.01% w / w to 1% w / w, 0.01% to 2% w / w, 0.1% w / w to 8% w / w, 0.1% w / w to 7% w / w, 0.1% w / w to 6% w / w, 0.1% w / w to 5% w / w, 0.1% w / w to 4% w / w, 0.1% w / w to 3% w / w, 0.1% w / w to 2% w / w, 2% w / w to 6% w / w, 2% w / w to 5% w / w, 2% w / w to 4% w / w, 3% w / w to 9% w / w, 5% w / w to 9% w / w, 7% w / w to 9% w / w, or 4% w / w to 6% w / w). In some embodiments, the resin has an approximate moisture content of 4.5% w / w to 6.5% w / w (e.g., 4.5% w / w, 4.6% w / w, 4.7% w / w, 4.8% w / w, 4.9% w / w, 5% w / w, 5.1% w / w, 5.2% w / w, 5.3% w / w, 5.4% w / w, 5.5% w / w, 5.6% w / w, 5.7% w / w, 5.8% w / w, 5.9% w / w, 6% w / w, 6.1% w / w, 6.2% w / w, 6.3% w / w, 6.4% w / w, or 6.5% w / w).

[0013] In some embodiments, the pre-passing step comprises passing a solvent through the resin to completely wet the resin. In some embodiments, the pre-passing step comprises passing the solvent through the resin in an amount of 1 bed volume (BV) to 4 BV (e.g., 1 BV, 2 BV, 3 BV, or 4 BV). In some embodiments, the pre-passing step comprises passing the solvent through the resin in an amount of 2 BV. In some embodiments, the solvent is passed through the resin in the pre-passing step with a minimum residence time in the column of at least 10 minutes. In some embodiments, the solvent is passed through the resin in the preliminary liquid passing step at a maximum downward flow superficial velocity of 0.5 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, or 1 cm / min to 3 cm / min). In some embodiments, the solvent is passed through the resin in the preliminary liquid passing step at a maximum downward flow superficial velocity of 1 cm / min to 4 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 2 cm / min to 4 cm / min, 3 cm / min to 4 cm / min, or 2 cm / min to 3 cm / min). In some embodiments, the solvent is passed through the resin at a maximum upflow superficial velocity of 0.05 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, 1 cm / min to 3 cm / min, 0.05 cm / min to 3 cm / min, 0.05 cm / min to 1 cm / min, or 0.05 cm / min to 0.1 cm / min). In some embodiments, the solvent is passed through the resin at a maximum upflow superficial velocity of 0.1 cm / min to 3 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 0.1 cm / min to 2 cm / min, 0.1 cm / min to 1 cm / min, or 2 cm / min to 3 cm / min). In some embodiments, the pre-passing step comprises thoroughly wetting the resin with the farnesene composition.

[0014] In some embodiments, the farnesene composition is exposed to the resin after the pre-passing step. In some embodiments, the farnesene composition is exposed to a resin that has not been subjected to a pre-passing step, which involves thoroughly wetting the resin by passing a solvent through the resin. In some embodiments, the farnesene composition is exposed to the resin for a minimum residence time on the resin of at least 10 minutes. In some embodiments, the farnesene composition is exposed to the resin at a maximum downward superficial velocity of 1 cm / min to 15 cm / min (e.g., 1 cm / min to 10 cm / min, 1 cm / min to 5 cm / min, 1 cm / min to 3 cm / min, 3 cm / min to 15 cm / min, 5 cm / min to 15 cm / min, 10 cm / min to 15 cm / min, or 5 cm / min to 10 cm / min). In some embodiments, the farnesene composition is exposed to the resin at a maximum downward flow superficial velocity of 5 cm / min to 11 cm / min (e.g., 5 cm / min, 6 cm / min, 7 cm / min, 8 cm / min, 9 cm / min, 10 cm / min, or 11 cm / min). In some embodiments, the farnesene composition is exposed to the resin at a maximum upward flow superficial velocity of 0.1 cm / min to 10 cm / min (e.g., 0.1 cm / min to 8 cm / min, 0.1 cm / min to 6 cm / min, 0.1 cm / min to 4 cm / min, 0.1 cm / min to 1 cm / min, 1 cm / min to 10 cm / min, 3 cm / min to 10 cm / min, 5 cm / min to 10 cm / min, 8 cm / min to 10 cm / min, 4 cm / min to 8 cm / min, or 2 cm / min to 6 cm / min). In some embodiments, the farnesene composition is exposed to the resin at a maximum upflow superficial velocity of 1 cm / min to 5 cm / min (e.g., 1 cm / min, 2 cm / min, 3 cm / min, 4 cm / min, or 5 cm / min). In some embodiments, steps (a)-(c) are repeated 2-20 times (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 times). The flow rate can be adjusted to result in higher purity isoprenoid product for different flow rates. For example, the flow rate can be reduced to result in higher purity isoprenoid product compared to when a higher flow rate is used. In some embodiments, steps (a)-(c) are repeated 2-10 times (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 times).In some embodiments, steps (a)-(c) are repeated 6 times. In some embodiments, steps (a)-(c) are repeated 2-4 times (e.g., 2, 3, or 4 times).

[0015] In some embodiments, the resin is regenerated after recovery of the farnesene from the resin in step (c). In some embodiments, the resin is regenerated by (a) adding a polar solvent to the resin, (b) removing the polar solvent from the resin, (c) heating the resin, and (d) cooling the resin. In some embodiments, step (a) is carried out by adding a polar solvent to the resin in an amount of 1 BV to 10 BV (e.g., 1 BV, 2 BV, 3 BV, 4 BV, 5 BV, 6 BV, 7 BV, 8 BV, 9 BV, or 10 BV). In some embodiments, the polar solvent is added to the resin in an amount of 2 BV to 6 BV (e.g., 1 BV, 2 BV, 3 BV, 4 BV, 5 BV, or 6 BV). In some embodiments, the polar solvent is added to the resin in an amount of about 4 BV. In some embodiments, step (c) is performed by heating the resin to a temperature of 100°C to 200°C (e.g., 100°C to 180°C, 100°C to 160°C, 100°C to 140°C, 100°C to 120°C, 120°C to 200°C, 150°C to 200°C, 170°C to 200°C, 140°C to 180°C, or 120°C to 160°C). In some embodiments, the resin is heated to a temperature of about 150°C. In some embodiments, the polar solvent is methanol. In some embodiments, the polar solvent is isopropanol. In some embodiments, the cooling in step (d) is performed by cooling the resin to room temperature. In some embodiments, the chromatography is performed under N2.

[0016] In some embodiments, the farnesene is purified from the farnesene composition to a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or more, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more). In some embodiments, the farnesene is purified from the farnesene composition at a purity of about 98% (w / w) to about 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98. Purified to 8% (w / w), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w). In some embodiments, the farnesene is purified from the farnesene composition to a purity of about 99% (w / w) to about 100% (w / w) (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the farnesene is purified from the farnesene composition to a purity of about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)).

[0017] In some embodiments, the farnesene has a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or greater, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or greater). In some embodiments, the farnesene has a purity of about 98% (w / w) to about 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98.8% (w / w), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 98.8% (w / w), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9 ...9% (w / w), 99.1% (w / w), 99. (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)), and optionally the farnesene has a purity of about 99% (w / w) to about 100% (w / w) (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the farnesene has a purity of about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the composition contains one or more impurities including farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0018] In certain embodiments, the present disclosure provides a composition comprising farnesene and one or more impurities, wherein the farnesene has a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or more, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more), and the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, HHFA, santalene, or santalol.

[0019] In some embodiments, the composition comprises farnesol and has a farnesol concentration of 0 mg / mL to 1 g / L (e.g., 0.2 g / L to 1 g / L, 0.4 g / L to 1 g / L, 0.6 g / L, 0.6 g / L to 1 g / L, 0.8 g / L to 1 g / L, 0 mg / mL to 0.8 g / L, 0 mg / mL to 0.6 g / L, 0 mg / mL to 0.4 g / L, 0 mg / mL to 0.2 g / L, 0.4 g / L to 0.6 g / L, or 0.2 g / L to 0.8 g / L).

[0020] In some embodiments, the concentration of farnesol is 0 mg / L to 600 mg / L (e.g., 0 mg / mL to 500 mg / mL, 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 600 mg / mL, 200 mg / mL to 600 mg / mL, 300 mg / mL to 600 mg / mL, 400 mg / mL to 600 mg / mL, 500 mg / mL to 600 mg / mL, 200 mg / mL to 400 mg / mL, or 300 mg / mL to 500 mg / mL). In some embodiments, the concentration of farnesol is less than 500 mg / L.

[0021] In some embodiments, the composition comprises farnesene epoxide and has a farnesol concentration of 0 mg / mL to 500 mg / L (e.g., 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 500 mg / mL, 200 mg / mL to 500 mg / mL, 300 mg / mL to 500 mg / mL, 400 mg / mL to 500 mg / mL, 200 mg / mL to 400 mg / mL, or 100 mg / mL to 300 mg / mL). In some embodiments, the concentration of the farnesene epoxide is between 0 mg / L and 200 mg / L (e.g., between 0 mg / L and 150 mg / L, between 0 mg / L and 100 mg / L, between 0 mg / L and 50 mg / L, between 50 mg / L and 200 mg / L, between 100 mg / L and 200 mg / L, between 150 mg / L and 200 mg / L, between 20 mg / L and 100 mg / L, or between 50 mg / L and 150 mg / L). In some embodiments, the concentration of the farnesene epoxide is less than 150 mg / L.

[0022] In certain aspects, the present disclosure provides compositions comprising farnesene and one or more carriers, diluents, or excipients, wherein the purity of the farnesene is about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)).

[0023] In some embodiments, the farnesene is present with one or more impurities, wherein the one or more impurities are present at a concentration of about 0.5% (w / w) or less.

[0024] In some embodiments, the one or more impurities are present at a concentration of about 0.4% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.3% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.2% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.1% (w / w) or less.

[0025] In some embodiments, the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0026] In some aspects, the present disclosure provides a method for purifying an isoprenoid, the method comprising: (a) providing an isoprenoid composition; and (b) purifying the isoprenoid from the isoprenoid composition of (a) by chromatography. In some embodiments, the isoprenoid composition comprises an isoprenoid and one or more impurities. In some embodiments, the one or more impurities comprise one or more polar impurities. In some embodiments, the one or more polar impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, HHFA, santalene, or santalol. In some embodiments, the isoprenoid composition has a concentration of one or more impurities between 1 ppm and 15,000 ppm (e.g., between 1 ppm and 10,000 ppm, between 1 ppm and 5,000 ppm, between 1 ppm and 1,0000 ppm, between 1 ppm and 500 ppm, between 1 ppm and 100 ppm, between 100 ppm and 15,000 ppm, between 1,000 ppm and 15,000 ppm, between 5,000 ppm and 15,000 ppm, between 10,000 ppm and 15,000 ppm, between 5,000 ppm and 12,000 ppm, between 1,000 ppm and 10,000 ppm, or between 3,000 ppm and 14,000 ppm). In some embodiments, the isoprenoid composition has a concentration of one or more impurities between 1000 ppm and 12,000 ppm (e.g., between 1000 ppm and 10,000 ppm, between 1000 ppm and 8,000 ppm, between 1000 ppm and 5,000 ppm, between 1000 ppm and 2,000 ppm, between 2,000 ppm and 12,000 ppm, between 5,000 ppm and 12,000 ppm, between 8,000 ppm and 12,000 ppm, between 10,000 ppm and 12,000 ppm, between 5,000 ppm and 10,000 ppm, or between 3,000 ppm and 10,000 ppm). In some embodiments, the isoprenoid composition has a concentration of one or more impurities between 100 ppm and 1,000 ppm.

[0027] In some embodiments, 4-tert-butylcatechol is added to the isoprenoid composition. In some embodiments, the 4-tert-butylcatechol is added to the isoprenoid composition to a concentration of 50 ppm to 150 ppm (e.g., 50 ppm to 125 ppm, 50 ppm to 100 ppm, 50 ppm to 75 ppm, 75 ppm to 150 ppm, 100 ppm to 150 ppm, 125 to 150 ppm, or 25 ppm to 75 ppm), and optionally, the 4-tert-butylcatechol is added to the isoprenoid composition to a concentration of about 100 ppm. In some embodiments, the chromatography includes (a) pre-flowing a resin, (b) exposing the isoprenoid composition to the resin, and (c) recovering the isoprenoid from the resin. In some embodiments, the resin includes aluminum oxide. In some embodiments, the aluminum oxide is basic aluminum oxide. In some embodiments, the aluminum oxide is acidic aluminum oxide. In some embodiments, the aluminum oxide is neutral aluminum oxide. In some embodiments, the resin includes silica. In some embodiments, the resin has a bulk density of 0.1 g / mL to 0.75 g / mL (e.g., 0.1 g / mL to 0.5 g / mL, 0.1 g / mL to 0.25 g / mL, 0.2 g / mL to 0.75 g / mL, 0.5 g / mL to 0.75 g / mL, 0. g / mL to 0.75 g / mL, or 0.3 g / mL to 0.5 g / mL). In some embodiments, the resin has a bulk density of 0.25 g / mL to 0.5 g / mL (e.g., 0.25 g / mL to 0.4 g / mL, 0.25 g / mL to 0.3 g / mL, 0.3 g / mL to 0.5 g / mL, 0.4 g / mL to 0.5 g / mL, or 0.3 g / mL to 0.4 g / mL). In some embodiments, the resin has a pore volume of 0.1 mL / g to 1.5 mL / g (e.g., 0.1 mL / g to 1.2 mL / g, 0.1 mL / g to 1 mL / g, 0.1 mL / g to 0.7 mL / g, 0.1 mL / g to 0.5 mL / g, 0.1 mL / g to 0.2 mL / g, 0.2 mL / g to 1.5 mL / g, 0.7 mL / g to 1.5 mL / g, 1 mL / g to 1.5 mL / g, 1.2 mL / g to 1.5 mL / g, or 0.5 mL / g to 1 mL / g).In some embodiments, the resin has a pore volume of 0.5 mL / g to 1 mL / g (e.g., 0.5 mL / g, 0.6 mL / g, 0.7 mL / g, 0.8 mL / g, 0.9 mL / g, or 1 mL / g).

[0028] In some embodiments, the resin has a particle distribution size of 25 μm to 800 μm (e.g., 25 μm to 700 μm, 25 μm to 600 μm, 25 μm to 500 μm, 25 μm to 400 μm, 25 μm to 300 μm, 25 μm to 200 μm, 25 μm to 100 μm, 25 μm to 50 μm, 50 μm to 100 μm, 50 μm to 800 μm, 100 μm to 800 μm, 200 μm to 800 μm, 300 μm to 800 μm, 500 μm to 800 μm, 600 μm to 800 μm, or 50 μm to 200 μm). In some embodiments, the resin has a particle distribution size of 100 μm to 800 μm (e.g., about 100 μm to about 700 μm, about 100 μm to about 600 μm, about 100 μm to about 500 μm, about 100 μm to about 400 μm, about 100 μm to about 300 μm, about 100 μm to about 200 μm, about 200 μm to about 800 μm, about 300 μm to about 800 μm, about 400 μm to about 800 μm, about 500 μm to about 800 μm, about 600 μm to about 800 μm, about 700 μm to about 800 μm, about 200 μm to about 600 μm). In some embodiments, the resin has a particle distribution size of 300 μm to 600 μm (e.g., about 300 μm to about 550 μm, about 300 μm to about 500 μm, about 300 μm to about 450 μm, about 300 μm to about 400 μm, about 300 μm to about 350 μm, about 350 μm to about 600 μm, about 400 μm to about 600 μm, about 450 μm to about 600 μm, about 500 μm to about 600 μm, about 550 μm to about 600 μm, or about 400 μm to about 500 μm). In some embodiments, the resin has an approximate moisture content of less than 0.01% w / w. In some embodiments, the resin has an approximate moisture content of 0.01% (w / w) to 9% (w / w) (e.g., 0.01% w / w to 1% w / w, 0.01% to 2% w / w, 0.1% w / w to 8% w / w, 0.1% w / w to 7% w / w, 0.1% w / w to 6% w / w, 0.1% w / w to 5% w / w, 0.1% w / w to 4% w / w, 0.1% w / w to 3% w / w, 0.1% w / w to 2% w / w, 2% w / w to 6% w / w, 2% w / w to 5% w / w, 2% w / w to 4% w / w, 3% w / w to 9% w / w, 5% w / w to 9% w / w, 7% w / w to 9% w / w, or 4% w / w to 6% w / w).In some embodiments, the resin has an approximate moisture content of 4.5% (w / w) to 6.5% (w / w) (e.g., 4.5% w / w, 4.6% w / w, 4.7% w / w, 4.8% w / w, 4.9% w / w, 5% w / w, 5.1% w / w, 5.2% w / w, 5.3% w / w, 5.4% w / w, 5.5% w / w, 5.6% w / w, 5.7% w / w, 5.8% w / w, 5.9% w / w, 6% w / w, 6.1% w / w, 6.2% w / w, 6.3% w / w, 6.4% w / w, or 6.5% w / w).

[0029] In some embodiments, the pre-passing step comprises passing a solvent through the resin to completely wet the resin. In some embodiments, the pre-passing step comprises passing 1 BV to 4 BV (e.g., 1 BV, 2 BV, 3 BV, or 4 BV) of the solvent through the resin. In some embodiments, the pre-passing step comprises passing 2 BV of the solvent through the resin. In some embodiments, the solvent is passed through the resin in the pre-passing step with a minimum residence time in the column of at least 10 minutes. In some embodiments, the solvent is passed through the resin in the preliminary liquid passing step at a maximum downward flow superficial velocity of 0.5 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, or 1 cm / min to 3 cm / min). In some embodiments, the solvent is passed through the resin in the preliminary liquid passing step at a maximum downward flow superficial velocity of 1 cm / min to 4 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 2 cm / min to 4 cm / min, 3 cm / min to 4 cm / min, or 2 cm / min to 3 cm / min). In some embodiments, the solvent is passed through the resin at a maximum upflow superficial velocity of 0.05 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, 1 cm / min to 3 cm / min, 0.05 cm / min to 3 cm / min, 0.05 cm / min to 1 cm / min, or 0.05 cm / min to 0.1 cm / min). In some embodiments, the solvent is passed through the resin at a maximum upward superficial velocity of 0.1 cm / min to 3 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 0.1 cm / min to 2 cm / min, 0.1 cm / min to 1 cm / min, or 2 cm / min to 3 cm / min). In some embodiments, the pre-passing step comprises thoroughly wetting the resin with the isoprenoid composition.

[0030] In some embodiments, the isoprenoid composition is exposed to the resin after the pre-passing step. In some embodiments, the isoprenoid composition is exposed to a resin that has not been subjected to a pre-passing step, which involves completely wetting the resin by passing a solvent through the resin. In some embodiments, the isoprenoid composition is exposed to the resin for a minimum residence time on the resin of at least 10 minutes. In some embodiments, the isoprenoid composition is exposed to the resin at a maximum downward superficial velocity of 1 cm / min to 15 cm / min (e.g., 1 cm / min to 10 cm / min, 1 cm / min to 5 cm / min, 1 cm / min to 3 cm / min, 3 cm / min to 15 cm / min, 5 cm / min to 15 cm / min, 10 cm / min to 15 cm / min, or 5 cm / min to 10 cm / min). In some embodiments, the isoprenoid composition is exposed to the resin at a maximum downward flow superficial velocity of 5 cm / min to 11 cm / min (e.g., 5 cm / min, 6 cm / min, 7 cm / min, 8 cm / min, 9 cm / min, 10 cm / min, or 11 cm / min). In some embodiments, the isoprenoid composition is exposed to the resin at a maximum upward flow superficial velocity of 0.1 cm / min to 10 cm / min (e.g., 0.1 cm / min to 8 cm / min, 0.1 cm / min to 6 cm / min, 0.1 cm / min to 4 cm / min, 0.1 cm / min to 1 cm / min, 1 cm / min to 10 cm / min, 3 cm / min to 10 cm / min, 5 cm / min to 10 cm / min, 8 cm / min to 10 cm / min, 4 cm / min to 8 cm / min, or 2 cm / min to 6 cm / min). In some embodiments, the isoprenoid composition is exposed to the resin at a maximum upflow superficial velocity of 1 cm / min to 5 cm / min (e.g., 1 cm / min, 2 cm / min, 3 cm / min, 4 cm / min, or 5 cm / min). In some embodiments, steps (a)-(c) are repeated 2-20 times (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 times). In some embodiments, steps (a)-(c) are repeated 2-10 times (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 times). In some embodiments, steps (a)-(c) are repeated 6 times. In some embodiments, steps (a)-(c) are repeated 2-4 times (e.g., 2, 3, or 4 times).

[0031] In some embodiments, the resin is regenerated after recovery of the isoprenoid from the resin in step (c). In some embodiments, the resin is regenerated by (a) adding a polar solvent to the resin, (b) removing methanol from the resin, (c) heating the resin, and (d) cooling the resin. In some embodiments, step (a) is carried out by adding a polar solvent to the resin in an amount of 1 BV to 10 BV (e.g., 1 BV, 2 BV, 3 BV, 4 BV, 5 BV, 6 BV, 7 BV, 8 BV, 9 BV, or 10 BV). In some embodiments, the polar solvent is added to the resin in an amount of 2 BV to 6 BV (e.g., 1 BV, 2 BV, 3 BV, 4 BV, 5 BV, or 6 BV). In some embodiments, the polar solvent is added to the resin in an amount of about 4 BV. In some embodiments, step (c) is performed by heating the resin to a temperature of 100°C to 200°C (e.g., 100°C to 180°C, 100°C to 160°C, 100°C to 140°C, 100°C to 120°C, 120°C to 200°C, 150°C to 200°C, 170°C to 200°C, 140°C to 180°C, or 120°C to 160°C). In some embodiments, the resin is heated to a temperature of about 150°C. In some embodiments, the polar solvent is methanol. In some embodiments, the polar solvent is isopropanol. In some embodiments, the cooling in step (d) is performed by cooling the resin to room temperature. In some embodiments, the chromatography is performed under N2.

[0032] In some embodiments, the isoprenoid is C5-C 60 Isoprenoids (e.g., C5-C 50 Isoprenoids, C5-C 40 Isoprenoids, C5-C 30 Isoprenoids, C5-C 20 Isoprenoids, C5-C 10 Isoprenoids, C 10 -C 60 Isoprenoids, C 20 -C 60 Isoprenoids, C30 -C 60 Isoprenoids, C 40 -C 60 Isoprenoids, C 50 -C 60 Isoprenoids, C 10 -C 40 Isoprenoids, C 20 -C 50 Isoprenoids, C 20 -C 30 Isoprenoids, C 15 -C 30 Isoprenoids, or C 30 -C 50 In some embodiments, the isoprenoid is a C 15 -C 60 Isoprenoids (e.g., C 15 -C 50 Isoprenoids, C 15 -C 40 Isoprenoids, C 15 -C 30 Isoprenoids, C 15 -C 20 Isoprenoids, C 20 -C 60 Isoprenoids, C 30 -C 60 Isoprenoids, C 40 -C 60 Isoprenoids, C 50 -C 60 Isoprenoids, C 25 -C 40 Isoprenoids, C 30 -C 50 Isoprenoids, C 20 -C 40 Isoprenoids, C 20 -C 50 Isoprenoids, C 20 -C 30 Isoprenoids, C 15 -C 30 Isoprenoids, or C 30 -C 50In some embodiments, the isoprenoid is a hemiterpenoid, monoterpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, tetraterpenoid, or polyterpenoid. In some embodiments, the isoprenoid is a sesquiterpenoid. In some embodiments, the isoprenoid is a hemiterpene, monoterpene, sesquiterpene, diterpene, sesterterpene, triterpene, tetraterpene, or polyterpene. In some embodiments, the isoprenoid is a sesquiterpene. In some embodiments, the isoprenoid is a monoterpenoid. In some embodiments, the isoprenoid is abietadiene, amorphadiene, cadinane, carene, cuminaldehyde, eugenol, farnesene, geranial, isoprene, limonene, myrcene, ocimene, α-pinene, β-pinene, sabinene, γ-terpinene, terpinolene, thujone, neral, eucalyptol, citronellal, carvone, or valencene. In some embodiments, the isoprenoid is purified from the isoprenoid composition to a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or more, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more). In some embodiments, the isoprenoid is purified from the isoprenoid composition at a purity of about 98% (w / w) to about 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98 Purified to 0.8% (w / w), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w).In some embodiments, the isoprenoid is purified from the isoprenoid composition to a purity of about 99% (w / w) to about 100% (w / w) (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the isoprenoid is purified from the isoprenoid composition to a purity of about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)).

[0033] In some aspects, the present disclosure provides a composition comprising an isoprenoid, the composition being produced by any one of the methods described herein. In some embodiments, the isoprenoid has a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or more, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more).

[0034] In some embodiments, the isoprenoid is about 98% (w / w) to about 100% (w / w) pure (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98.8% (w / w), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99 0.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)), and optionally the isoprenoid is purified from the isoprenoid composition to a purity of about 99% (w / w) to about 100% (w / w) (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the isoprenoid has a purity of about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the composition contains one or more impurities including farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acids, sterols, HHFA, santalene, or santalol.

[0035] In certain aspects, the disclosure provides a composition comprising an isoprenoid and one or more impurities, wherein the isoprenoid has a purity of about 90% (w / w) to about 100% (w / w), and the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, an HHFA, santalene, or santalol. In some embodiments, the composition comprises farnesol and has a farnesol concentration of 0 mg / mL to 1 g / L (e.g., 0.2 g / L to 1 g / L, 0.4 g / L to 1 g / L, 0.6 g / L, 0.6 g / L to 1 g / L, 0.8 g / L to 1 g / L, 0 mg / mL to 0.8 g / L, 0 mg / mL to 0.6 g / L, 0 mg / mL to 0.4 g / L, 0 mg / mL to 0.2 g / L, 0.4 g / L to 0.6 g / L, or 0.2 g / L to 0.8 g / L). In some embodiments, the concentration of farnesol is 0 mg / L to 600 mg / L (e.g., 0 mg / mL to 500 mg / mL, 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 600 mg / mL, 200 mg / mL to 600 mg / mL, 300 mg / mL to 600 mg / mL, 400 mg / mL to 600 mg / mL, 500 mg / mL to 600 mg / mL, 200 mg / mL to 400 mg / mL, or 300 mg / mL to 500 mg / mL). In some embodiments, the concentration of farnesol is less than 500 mg / L.

[0036] In some embodiments, the composition comprises farnesene epoxide and has a farnesol concentration of 0 mg / mL to 500 mg / L (e.g., 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 500 mg / mL, 200 mg / mL to 500 mg / mL, 300 mg / mL to 500 mg / mL, 400 mg / mL to 500 mg / mL, 200 mg / mL to 400 mg / mL, or 100 mg / mL to 300 mg / mL). In some embodiments, the concentration of the farnesene epoxide is between 0 mg / L and 200 mg / L (e.g., between 0 mg / L and 150 mg / L, between 0 mg / L and 100 mg / L, between 0 mg / L and 50 mg / L, between 50 mg / L and 200 mg / L, between 100 mg / L and 200 mg / L, between 150 mg / L and 200 mg / L, between 20 mg / L and 100 mg / L, or between 50 mg / L and 150 mg / L). In some embodiments, the concentration of the farnesene epoxide is less than 150 mg / L.

[0037] In some aspects, the disclosure provides compositions comprising an isoprenoid and one or more carriers, diluents, or excipients, wherein the isoprenoid has a purity of about 99.5% (w / w) to about 100% (w / w) (e.g., 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the isoprenoid is present with one or more impurities, wherein the one or more impurities are present at a concentration of about 0.5% (w / w) or less.

[0038] In some embodiments, the one or more impurities are present at a concentration of about 0.4% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.3% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.2% (w / w) or less. In some embodiments, the one or more impurities are present at a concentration of about 0.1% (w / w) or less. In some embodiments, the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, HHFA, santalene, or santalol.

[0039] definition As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0040] The term "about" when used herein to modify a numerical value or range includes normal variations encountered in the art and includes the numerical value or the endpoints of the numerical range plus or minus 1 to 10% (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%). Thus, a value of 10 includes all numbers from 9 to 11. All numerical ranges described herein include, to the first significant digit, the endpoints of the range, and all numbers between the endpoints, unless otherwise specified.

[0041] As used herein, "acidic aluminum oxide" refers to an amphoteric oxide having a chemical formula of Al2O3 that has a pH of about ≦4.5 when exposed to an aqueous environment (e.g., when contacted with water).

[0042] As used herein, "basic aluminum oxide" refers to an amphoteric oxide having a chemical formula of Al2O3 that has a pH of about ≥ 10 when exposed to an aqueous environment (e.g., when contacted with water).

[0043] As used herein, the term "bulk density" refers to the weight per volume unit of a powder, typically expressed in g / cm 3 , kg / m 3, g / 100 mL, or g / mL. Bulk density is usually determined by measuring the volume of 100 g of powder in a 250 ml graduated cylinder after subjecting it to standardized tapping compression.

[0044] As used herein, the term "capable of producing" refers to a host cell that contains the enzymes necessary to produce a given compound according to the biochemical pathway that produces that compound. For example, a cell (e.g., a yeast cell) that is "capable of producing" an isoprenoid is one that contains the enzymes necessary to produce an isoprenoid according to the isoprenoid biosynthetic pathway.

[0045] As used herein, the term "evaporation" describes a process in which at least a portion of a liquid undergoes a change of state and assumes a gaseous state. For example, evaporation may be used to separate two liquids from one another or to remove one liquid from a mixture containing one or more additional liquids. In some embodiments, evaporation includes the process of distillation, in which a liquid not only changes phase to a gaseous state but is then condensed back into a liquid state. In some embodiments of the present disclosure, distillation is performed by heating a mixture of liquids so that lower-boiling substances begin to evaporate, changing phase from a liquid to a gaseous state while leaving the liquid(s) remaining in the mixture in a liquid phase. The lower-boiling substances can then be condensed, for example, after exposure to low temperatures, thereby (1) returning the lower-boiling substances to a liquid phase and (2) separating the lower-boiling substances from the liquid(s) remaining in the mixture.

[0046] Distillation may be, for example, "simple distillation," which refers to a process in which a mixture of liquids having substantially different boiling points (e.g., boiling points that differ from each other by more than about 25°C) is separated by heating the mixture until substantially all of the lower-boiling material is vaporized and substantially all of the higher-boiling material remains in the liquid phase. The vapor of the lower-boiling material is then condensed, returning the lower-boiling material to the liquid phase. In some embodiments, distillation may be "fractional distillation," i.e., a process used to separate, for example, highly miscible liquids and / or liquids whose boiling points differ by less than about 25°C. Fractional distillation processes involve heating a mixture containing the liquids so that the resulting vapor enters a fractionation column. The fractionation column is a column (e.g., a vertical column, a tilted column, or a horizontal column) configured to have a temperature gradient, with the bottom of the fractionation column (i.e., the point where the vapor enters the column) being the warmest and the top of the fractionation column being the coldest. As the vapor travels upward through the column, it becomes enriched in lower boiling materials as a result of the temperature gradient. After being enriched in lower boiling materials, the vapor exits the fractionation tower and is exposed to lower temperatures, causing the lower boiling materials to condense and separate the liquid mixture into its components.

[0047] As used herein, the term "pharmaceutical composition" refers to a mixture containing therapeutic or prophylactic compounds that is administered to a subject, e.g., a mammal, e.g., a human, to prevent, treat, or control a particular disease or condition that affects or can affect the mammal.

[0048] As used herein, the term "farnesene composition" refers to a mixture (e.g., a solution or suspension) comprising farnesene. In some embodiments, the farnesene composition comprises farnesene and one or more impurities.

[0049] As used herein, "neutral aluminum oxide" refers to an amphoteric oxide that has the chemical formula Al2O3 and has a pH of about 7 when exposed to an aqueous environment (e.g., when contacted with water).

[0050] As used herein, the term "pharmaceutically acceptable" refers to compounds, substances, compositions and / or dosage forms that are suitable for contact with the tissues of a subject, such as a mammal (e.g., a human), without undue toxicity, irritation, allergic response, or other adverse complications, commensurate with a reasonable benefit / risk ratio.

[0051] As used herein, the term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt of a compound described herein. For example, a pharmaceutically acceptable salt of any compound described herein includes a salt that, within the scope of sound medical judgment, is suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, or allergic reaction, and that is commensurate with a reasonable benefit / risk ratio. Examples of pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Such salts can be prepared in situ, for example, during the final isolation and purification process of a compound described herein, or can be prepared independently by reacting a free base group with a suitable organic acid.

[0052] The compounds described herein may have ionic groups that can be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts incorporating inorganic or organic acids, or these salts may be prepared from inorganic or organic bases when the compounds described herein are in acidic form. The compounds may be prepared or used as pharmaceutically acceptable salts synthesized as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable salts may be prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic and organic acids and bases. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, and 2-hydroxyethanesulfonate. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium salts, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0053] As used herein, the term "polar solvent" refers to a solvent that has charge separation and the ability to solvate other polar substances, including, but not limited to, acetone, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), isopropanol, and methanol.

[0054] As used herein, the term "pore volume" or "porosity" refers to a measure of the voids in a material. These parameters are generally measured as a volume percentage. Volume percentage can be expressed as the volume of voids in a material relative to the total volume of the material. Pore volume is typically expressed in mL / g cm 3 / g, or m 3 It is expressed in / kg.

[0055] As used herein, the term "production" generally refers to the amount of a compound produced by a host cell provided herein. In some embodiments, production is expressed as the yield of the compound by the host cell. In other embodiments, production is expressed as the productivity of the host cell in producing the compound.

[0056] As used herein, the term "residence time" refers to the total time a material (e.g., a solvent or isoprenoid composition described herein, e.g., a farnesene composition) remains on a column in a column chromatography procedure. A material (e.g., a solvent or isoprenoid composition described herein, e.g., a farnesene composition) having a faster flow rate will have a shorter residence time compared to a material (e.g., a solvent or isoprenoid composition described herein, e.g., a farnesene composition) having a slower flow rate. Similarly, a material (e.g., a solvent or isoprenoid composition described herein, e.g., a farnesene composition) having a slower flow rate will have a longer residence time compared to a material (e.g., a solvent or isoprenoid composition described herein, e.g., a farnesene composition) having a faster flow rate.

[0057] As used herein, the term "silica" refers to a silicon dioxide (SiO2)-based resin material that can be used, for example, in column chromatography procedures. The term "silica" encompasses, inter alia, amorphous forms of silica consisting of an irregular three-dimensional structure, which are porous materials with microscale to nanoscale pores.

[0058] As used herein, the terms "subject" and "patient" are synonymous and refer to an organism receiving therapeutic or prophylactic treatment for a particular disease or condition described herein. Examples of subjects and patients include mammals, such as humans.

[0059] As used herein in the context of column chromatography procedures, the term "superficial velocity" refers to the volumetric flow rate of a fluid divided by the cross-sectional area of ​​the empty column through which the fluid passes. Superficial velocity is therefore not affected by the presence or structure of a packed bed, nor does it change whether the particles within the bed are porous or non-porous.

[0060] As used herein, the term "moisture content" refers to the amount of water bound to a resin and is characterized in terms of the percentage of the resin's weight that comes from the bound water. The amount of moisture absorbed by a resin can depend on a variety of factors, including the type of resin, ambient temperature, seasonal variations, packaging and shipping issues, and manufacturing.

[0061] As used herein, the term "wetting" refers to contacting the resin with a solvent containing dissolved material for isolation. For example, in some embodiments, "wetting" occurs when a mixture containing isoprenoid compounds is contacted with the resin. [Brief explanation of the drawings]

[0062] [Figure 1] The components of the resulting high purity farnesene product are shown below. [Figure 2A]1 is a graph showing the results of purity of farnesene isolated by removal of farnesene epoxide, farnesol, and farneseic acid by alumina stripping using basic alumina from Sorbent Technology. [Figure 2B] 1 is a graph showing the results of purity of farnesene isolated by removal of farnesene epoxide, farnesol, and farneseic acid by alumina stripping using basic alumina from Sigma Aldrich. [Figure 2C] 1 is a graph showing the results of purity of farnesene isolated by removal of farnesene epoxide, farnesol, and farneseic acid by alumina stripping using Activity Super I activated alumina from Sorbent Technology. [Figure 3] 1 is a graph showing breakthrough data for santalol after alumina column treatment, where over 12000 ppm of santalol was reduced to below detection levels in the first 3 bed volumes (BV). [Figure 4] 1 is a graph showing breakthrough data for hexahydrofarneseic acid (HHFA) after alumina column treatment, with no HHFA observed in the first 2 BV. [Figure 5] 1 is a graph showing the concentration of products over time after treating farnesene with H3PO4. [Figure 6] Graphs A and B show the change over time in (i) farnesol and farnesene epoxide concentrations (A) and (ii) farnesene and farnesene dimer (B) in distilled farnesene at 25°C, 57.5°C, and 90°C and at acid concentrations of 2 wt%, 6 wt%, and 10 wt%. [Figure 7] 1 is a graph showing solution color over time of distilled farnesene after treatment with acid at concentrations of 2 wt %, 6 wt %, and 10 wt % at 25° C., 57.5° C., and 90° C. [Figure 8]1A and 1B are graphs showing the amount of dimer formation and farnesene loss (B) during the treatment of (i) (A) and (ii) crude farnesene with H3PO4. DETAILED DESCRIPTION OF THE INVENTION

[0063] The present disclosure provides a method for isolating isoprenoids so that the resulting isoprenoids are highly pure. The isoprenoids can be isolated from any source, for example, from fermentation compositions, plants, or animals. Considering the goal of producing and isolating highly pure isoprenoids, the challenges in purifying isoprenoids to a high degree have been significant. For example, isoprenoids purified using conventional distillation techniques result in isoprenoid compositions of insufficient purity for certain applications, such as polymerization reactions.

[0064] It has now been discovered that the combination of a unique series of chromatography steps can result in isoprenoid compositions with excellent purity levels (e.g., purity levels of 99.5% (w / w) or greater). For example, the methods and compositions described herein can be used to purify isoprenoids from compositions containing the isoprenoids. To recover highly pure isoprenoids from the isoprenoid composition, the composition can be subjected to one or more chromatography steps. In these steps, the isoprenoid composition is contacted with a resin at a specific flow rate. The resin is prepared and regenerated after contact with the isoprenoid solution. Repetition of these steps results in a highly pure isoprenoid product suitable for use in, for example, polymerization reactions. The present disclosure is based, at least in part, on the applicant's discovery that the chromatography steps described herein were capable of efficiently removing very high levels of impurities (e.g., >10,000 ppm) from the isoprenoid. This is in contrast to other tested methods that result in higher levels of impurities.

[0065] The following sections describe exemplary compositions and methods that can be used to perform the chromatography steps of the present disclosure.

[0066] Isoprenoids Described herein are methods for purifying isoprenoids, isolating isoprenoids from isoprenoid sources, and producing isoprenoids with high purity. The isoprenoids of the present disclosure are C5-C 60 Isoprenoids (e.g., C5 isoprenoids, C 10 Isoprenoids, C 15 Isoprenoids, C 20 Isoprenoids, C 25 Isoprenoids, C 30 Isoprenoids, C 35 Isoprenoids, C 40 Isoprenoids, C 45 Isoprenoids, C 50 Isoprenoids, C 55 Isoprenoids, or C 60 For example, in some embodiments, the isoprenoid can be a C 20 In some embodiments, the isoprenoid is a C 60 These compounds are derived from four isoprene units and are also called diterpenoids. Specific examples of diterpenoids include casbene, eleutherobin, paclitaxel, prostratin, pseudopterosin, taxadiene, and salvinorin. In yet another example, the isoprenoid is C 20+ These compounds are derived from more than four isoprene units, while triterpenoids (derived from six isoprene units) are 30 isoprenoid compounds), such as arbrusidee, bruceanthin, testosterone, progesterone, cortisone, digitoxin, and squalene, tetraterpenoids (C40 isoprenoid compounds obtained from eight isoprenoids), such as β-carotene, and polyterpenoids (C40 isoprenoid compounds obtained from more than eight isoprene units), such as β-carotene. 40+isoprenoid compounds), such as polyisoprene. In some embodiments, the isoprenoid is selected from the group consisting of abietadiene, amorphadiene, cadinane, carene, cuminaldehyde, eugenol, farnesene, geranial, isoprene, limonene, myrcene, ocimene, α-pinene, β-pinene, sabinene, γ-terpinene, terpinolene, thujone, neral, eucalyptol, citronellal, carvone, and valencene. In some embodiments, the isoprenoid is cadinene. In some embodiments, the isoprenoid is farnesene.

[0067] The isoprenoid product is C5-C 20 Isoprenoids (e.g., C5 isoprenoids, C6 isoprenoids, C7 isoprenoids, C8 isoprenoids, C9 isoprenoids, C 10 Isoprenoids, C 11 Isoprenoids, C 12 Isoprenoids, C 13 Isoprenoids, C 14 Isoprenoids, C 15 Isoprenoids, C 16 Isoprenoids, C 17 Isoprenoids, C 18 Isoprenoids, C 19 Isoprenoids, or C 20 In some embodiments, the isoprenoid produced by the cells is a C5 isoprenoid. These compounds are derived from a single isoprene unit and are also called hemiterpenoids. Specific examples of hemiterpenoids are isoprene and isoprenol.

[0068] The isoprenoid product is C 10 -C 15 Isoprenoids (e.g., C 10 Isoprenoids, C 11 Isoprenoids, C 12 Isoprenoids, C 13 Isoprenoids, C 14 Isoprenoids, or C 15In other embodiments, the isoprenoid may be a C 10 These compounds are isoprenoids. These compounds are derived from two isoprene units and are also called monoterpenoids. Specific examples of monoterpenoids include limonene, citranellol, geraniol, menthol, perillyl alcohol, linalool, thujone, myrcene, hinokitiol, carvacrol, anethole, cuminaldehyde, eucalyptol, α-pinene, β-pinene, citronellal, isopulegol, nerol, neral, geranial, and carvone. In another embodiment, the isoprenoid is C 15 These compounds are derived from three isoprene units and are also called sesquiterpenoids. Specific examples of sesquiterpenoids are periplanone B, ginkgolide B, amorphadiene, artemisinin, artemisinic acid, valencene, nootkatone, epicedrol, epiaristroken, farnesol, gossypol, sanonin, periplanone, forskolin, and patchouli alcohol, also known as patchouliol.

[0069] Isoprenoid compounds also include, but are not limited to, carotenoids (e.g., lycopene, α- and β-carotene, α- and β-cryptoxanthin, bixin, zeaxanthin, astaxanthin, and lutein), steroid compounds, cannabinoids, and other chemical groups, such as mixed terpene-alkaloids, and compounds composed of isoprenoids modified by coenzyme Q (CoQ) enzymes, e.g., coenzyme Q-10.

[0070] In some embodiments, the isoprenoid is a hemiterpenoid, monoterpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, tetraterpenoid, or polyterpenoid. In some embodiments, the isoprenoid is a monoterpenoid.

[0071] terpenes In some embodiments, the isolated product is a terpene. In some embodiments, the terpene is a C5-C 60 Terpenes (e.g., C5 isoprenoids, C 10 Isoprenoids, C 15 Isoprenoids, C 20 Isoprenoids, C 25 Isoprenoids, C 30 Isoprenoids, C 35 Isoprenoids, C 40 Isoprenoids, C 45 Isoprenoids, C 50 Isoprenoids, C 55 Isoprenoids, or C 60 In some embodiments, the terpene is a C5-C 20 Terpenes (e.g., C5 terpenes, C6 terpenes, C7 terpenes, C8 terpenes, C9 terpenes, C 10 Terpene, C 11 Terpene, C 12 Terpene, C 13 Terpene, C 14 Terpene, C 15 Terpene, C 16 Terpene, C 17 Terpene, C 18 Terpene, C 19 Terpenes, or C 20 In some embodiments, the terpene is C 10 -C 15 Terpenes (e.g., C 10 Terpene, C 11 Terpene, C 12 Terpene, C 13 Terpene, C 14 Terpenes, or C 15 In some embodiments, the terpene is a hemiterpene, monoterpene, sesquiterpene, diterpene, sesterterpene, triterpene, tetraterpene, or polyterpene. In some embodiments, the terpene is a sesquiterpene.

[0072] In some embodiments, the terpene is a monoterpene.10 It is a terpene derived from two isoprene units. For example, the monoterpene may be carene, the structure of which is: [ka] Carene is typically produced from GPP by carene synthase. Specific examples of suitable nucleotide sequences for use as heterologous sequences encoding carene synthase include, but are not limited to, (AF461460, REGION 43...1926, Picea abies) and (AF527416, REGION:78...1871, Salvia stenophylla). Another monoterpene, such as geraniol (also known as rhodnol), has the structure [ka] and can be a product produced by the present invention. Geraniol is typically produced from OPP by geraniol synthase. Specific examples of suitable nucleotide sequences encoding geraniol synthases that can be used in the heterologous sequences of the present invention include, but are not limited to, (AJ457070, Cinnamomum tenuipilum), (AY362553, Ocimum basilicum), (DQ234300, Perilla frutescens strain 1864), (DQ234299, Perilla citriodora strain 1861), (DQ234298, Perilla citriodora strain 4935), and (DQ088667, Perilla citriodora). The monoterpene linalool, whose structure is [ka] is normally produced from GPP by linalool synthase and can be produced by the present invention. Specific examples of suitable nucleotide sequences include (AF497485, Arabidopsis thaliana), (AC002294, Locus AAB71482, Arabidopsis thaliana), (AY059757, Arabidopsis thaliana), (NM_104793, Arabidopsis thaliana), (AF154124, Artemisia annua), (AF067603, Clarkia breweri), (AF067602, Clarkia concinna), (AF067601, Clarkia breweri), (U58314, Clarkia breweri), (AY840091, Lycopersicon esculentum), (DQ263741, Lavandula angustifolia), (AY083653, Mentha citrate), (AY693647, Ocimum basilicum), (XM_463918, Oryza sativa), (AP004078, Locus BAD07605, Oryza sativa), (XM_463918, Locus XP_463918, Oryza sativa), (AY917193, Perilla citriodora), (AF271259, Perilla frutescens), (AY473623, Picea abies), (DQ195274, Picea sitchensis), and (AF444798, Perilla frutescens var. crispa cultivar No. 79). These sequences may be used as heterologous sequences in the present invention. Another monoterpene, limonene, has the structure [ka] and is normally produced from GPP by limonene synthase. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences of the present invention include, but are not limited to, (+)-limonene synthase (AF514287, REGION:47...1867, Citrus limon) and (AY055214, REGION:48...1889, Agastache rugosa), and (-)-limonene synthase (DQ195275, REGION:1...1905, Picea sitchensis), (AF006193, REGION:73.1986, Abies grandis), and (MC4SLSP, REGION:29...1828, Mentha spicata).

[0073] Myrcene, a monoterpene, has the structure [ka] , which is normally produced from GPP by myrcene synthase and is another product that can be produced by the present invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences of the present invention include, but are not limited to, (187908, Abies grandis), (AY195609, Antirrhinum majus), (AY195608, Antirrhinum majus), (NM_127982, Arabidopsis thaliana TPS10), NM_113485, Arabidopsis thaliana ATTPS-CIN), (NM_13483, Arabidopsis thaliana ATIPS-CIN), (AF271259, Perilla frutescens), (AY473626, Picea abies), (AF369919, Picea abies), and (AJ304839, Quercus ilex).

[0074] Another monoterpene, ocimene, i.e., α- and β-ocimene, has the structure [ka] and is normally produced from GPP by ocimene synthase, a synthase that can be encoded by a heterologous sequence of the present invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences include (AY195607, Antirrhinum majus), (AY195609, Antirrhinum majus), (AY195608, Antirrhinum majus), (AK221024, Arabidopsis thaliana), (NM_113485, Arabidopsis thaliana ATTPS-CIN), (NM_113483, Arabidopsis thaliana ATTPS-CIN), (NM_117775, Arabidopsis thaliana ATTPS03), (NM_001036574, Arabidopsis thaliana ATTPS03), (NM_127982, Arabidopsis thaliana TPS10), (AB110642, Citrus unshiu CitMTSL4), and (AY575970, Lotus corniculatus var. japonicus).

[0075] Another monoterpene, α-pinene, has the structure [ka] and is normally produced from GPP by α-pinene synthase, a synthase that can be encoded by a heterologous sequence of the invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences to encode the synthase include, but are not limited to, (+) α-pinene synthase (AF543530, REGION:1...1887, Pinus taeda), (-) α-pinene synthase (AF543527, REGION:32...1921, Pinus taeda), and (+) / (-) α-pinene synthase (AGU87909, REGION:6111892, Abies grandis).

[0076] Another monoterpene, β-pinene, has the structure [ka] and is normally produced from GPP by β-pinene synthase, a synthase that can be encoded by a heterologous sequence of the invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences to encode the synthase include, but are not limited to, (-)β-pinene synthase (AF276072, REGION:1...1749, Artemisia annua) and (AF514288, REGION:26...1834, Citrus limon).

[0077] Another monoterpene, sabinene, has the structure [ka] and is normally produced from GPP by sabinene synthase, a synthase that can be encoded by a heterologous sequence of the present invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences include, but are not limited to, AF051901, REGION:26...1798 from Salvia officinalis.

[0078] Another monoterpene, γ-terpinene, has the structure [ka] and is normally produced from GPP by gamma-terpinene synthase, a synthase that can be encoded by a heterologous sequence of the present invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences include, but are not limited to, (AF514286 from Citrus limon, REGION: 30...1832) and (AB110640 from Citrus unshiu, REGION 1...1803).

[0079] Another monoterpene, terpinolene, has the structure [ka] and is normally produced from GPP by terpinolene synthase, a synthase that can be encoded by a heterologous sequence of the invention. Specific examples of suitable nucleotide sequences that can be used as heterologous sequences include, but are not limited to, (AY693650 from Oscimum basilicum) and (AY906866 from Pseudotsuga menziesii, REGION:10...1887).

[0080] In some embodiments, the product is abietadiene, amorphadiene, carene, α-farnesene, β-farnesene, farnesol, geraniol, geranylgeraniol, isoprene, linalool, limonene, myrcene, nerolidol, ocimene, patchouliol, β-pinene, sabinene, γ-terpinene, terpinolene, menthol, eucalyptol, citronellol, citronellal, or valencene. In some embodiments, the isolated product is a cadinane, which has the following structure: [ka] It has.

[0081] In some embodiments, the isolated product is farnesene. In some embodiments, the product has the following structure: [ka] It is β-farnesene having the formula:

[0082] In some embodiments, the product is myrcene. In some embodiments, the product is pinene. In some embodiments, the product is limonene.

[0083] Methods for isolating isoprenoids

[0010] Described herein are methods for purifying isoprenoids, isolating isoprenoids from isoprenoid sources, and producing isoprenoids having high purity. The methods described herein can include, for example, optionally isolating farnesene from an evaporated (e.g., distilled) farnesene composition to recover the farnesene product in high purity. Exemplary possible chromatography steps are described in further detail in the following sections.

[0084] The isoprenoids can optionally be isolated from the evaporated isoprenoid composition (e.g., distilled isoprenoids). For example, the isoprenoids can be isolated from the evaporated isoprenoid composition obtained by distillation of a composition containing isoprenoids, and the composition is obtained by extraction. The distillation can be carried out using any procedure known in the art. In some embodiments, the distillation can be simple distillation. In some embodiments, the distillation can be fractional distillation.

[0085] The resulting isoprenoid composition may contain one or more impurities. The isoprenoid composition may contain the isoprenoid and one or more impurities. One or more of the impurities may be polar impurities. For example, the polar impurities may include farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acids, sterols, hexahydrofarneseic acid (HHFA), santalene, or santalol. The one or more impurities may be present in an amount of 1 ppm to 15,000 ppm (e.g., 1 ppm to 10,000 ppm, 1 ppm to 5,000 ppm, 1 ppm to 1,0000 ppm, 1 ppm to 500 ppm, 1 ppm to 100 ppm, 100 ppm to 15,000 ppm, 1,000 ppm to 15,000 ppm, 5,000 ppm to 15,000 ppm, 10,000 ppm to 15,000 ppm, 5,000 ppm to 12,000 ppm, 1,000 ppm to 10,000 ppm, or 3,000 ppm to 14,000 ppm). For example, the isoprenoid composition has a concentration of one or more impurities between 1000 ppm and 12,000 ppm (e.g., between 1000 ppm and 10,000 ppm, between 1000 ppm and 8,000 ppm, between 1000 ppm and 5,000 ppm, between 1000 ppm and 2,000 ppm, between 2,000 ppm and 12,000 ppm, between 5,000 ppm and 12,000 ppm, between 8,000 ppm and 12,000 ppm, between 10,000 ppm and 12,000 ppm, between 5,000 ppm and 10,000 ppm, or between 3,000 ppm and 10,000 ppm). In some embodiments, the isoprenoid composition has a concentration of one or more impurities of 100 ppm to 1,000 ppm (e.g., 100 ppm to 800 ppm, 100 ppm to 600 ppm, 100 ppm to 400 ppm, 100 ppm to 200 ppm, 200 ppm to 1,000 ppm, 500 ppm to 1,000 ppm, 800 ppm to 1,000 ppm, or 300 ppm to 800 ppm). In some embodiments, an antioxidant may be added to the composition. For example, the antioxidant may be 4-tert-butylcatechol.In some embodiments, the 4-tert-butylcatechol is added to the farnesene composition to a concentration of 50 ppm to 150 ppm (e.g., 50 ppm to 125 ppm, 50 ppm to 100 ppm, 50 ppm to 75 ppm, 75 ppm to 150 ppm, 100 ppm to 150 ppm, 125 ppm to 150 ppm, or 25 ppm to 75 ppm). In some embodiments, the 4-tert-butylcatechol is added to the farnesene composition to a concentration of about 100 ppm.

[0086] Chromatography Disclosed herein are methods for purifying an isoprenoid, the methods including providing an isoprenoid composition and purifying the isoprenoid from the isoprenoid composition by chromatography. For example, described herein are methods for purifying farnesene, the methods including providing a farnesene composition and purifying the farnesene from the farnesene composition by chromatography.

[0087] In some embodiments, chromatography is used to isolate isoprenoids, for example, following extraction and evaporation of the isoprenoids from the isoprenoid source of the present disclosure. In some embodiments, chromatography is used to isolate the isoprenoids from the isoprenoid composition. The chromatography can include pre-loading a resin, exposing the isoprenoid composition to the resin, and recovering the isoprenoid from the resin. The chromatography step can include, for example, exposing the isoprenoid composition to the resin and recovering the isoprenoid from the resin.

[0088] The resin can be an aluminum oxide resin, such as a basic aluminum oxide resin, an acidic aluminum oxide resin, or a neutral aluminum oxide resin. In some embodiments, the resin can be a silica resin. The particle size of the resin can be, for example, about 100 μm to about 800 μm (e.g., about 100 μm to about 700 μm, about 100 μm to about 600 μm, about 100 μm to about 500 μm, about 100 μm to about 400 μm, about 100 μm to about 300 μm, about 100 μm to about 200 μm, about 200 μm to about 800 μm, about 300 μm to about 800 μm, about 400 μm to about 800 μm, about 500 μm to about 800 μm, about 600 μm to about 800 μm, about 700 μm to about 800 μm, about 200 μm to about 600 μm).

[0089] For example, the particle size of the resin can be about 300 μm to about 600 μm (e.g., about 300 μm to about 550 μm, about 300 μm to about 500 μm, about 300 μm to about 450 μm, about 300 μm to about 400 μm, about 300 μm to about 350 μm, about 350 μm to about 600 μm, about 400 μm to about 600 μm, about 450 μm to about 600 μm, about 500 μm to about 600 μm, about 550 μm to about 600 μm, or about 400 μm to about 500 μm). The resin may require an activation step. The activation step can be, for example, drying the resin before use.

[0090] The resin may have a bulk density of 0.1 g / mL to 0.75 g / mL (e.g., 0.1 g / mL to 0.5 g / mL, 0.1 g / mL to 0.25 g / mL, 0.2 g / mL to 0.75 g / mL, 0.5 g / mL to 0.75 g / mL, 0.g / mL to 0.75 g / mL, or 0.3 g / mL to 0.5 g / mL). For example, the resin may have a bulk density of 0.25 g / mL to 0.5 g / mL (e.g., 0.25 g / mL to 0.4 g / mL, 0.25 g / mL to 0.3 g / mL, 0.3 g / mL to 0.5 g / mL, 0.4 g / mL to 0.5 g / mL, or 0.3 g / mL to 0.4 g / mL). In some embodiments, the resin has a pore volume of 0.1 mL / g to 1.5 mL / g (e.g., 0.1 mL / g to 1.2 mL / g, 0.1 mL / g to 1 mL / g, 0.1 mL / g to 0.7 mL / g, 0.1 mL / g to 0.5 mL / g, 0.1 mL / g to 0.2 mL / g, 0.2 mL / g to 1.5 mL / g, 0.7 mL / g to 1.5 mL / g, 1 mL / g to 1.5 mL / g, 1.2 mL / g to 1.5 mL / g, or 0.5 mL / g to 1 mL / g). For example, the resin can have a pore volume of 0.5 mL / g to 1 mL / g (e.g., 0.5 mL / g, 0.6 mL / g, 0.7 mL / g, 0.8 mL / g, 0.9 mL / g, or 1 mL / g). The resin may have an approximate moisture content of less than 1% w / w, hi some embodiments, the resin has an approximate moisture content of 2% to 9% w / w (e.g., 2% to 6% w / w, 2% to 5% w / w, 2% to 4% w / w, 3% to 9% w / w, 5% to 9% w / w, 7% to 9% w / w, or 4% to 6% w / w). For example, the resin has an approximate moisture content of 4.5% w / w to 6.5% w / w (e.g., 4.5% w / w, 4.6% w / w, 4.7% w / w, 4.8% w / w, 4.9% w / w, 5% w / w, 5.1% w / w, 5.2% w / w, 5.3% w / w, 5.4% w / w, 5.5% w / w, 5.6% w / w, 5.7% w / w, 5.8% w / w, 5.9% w / w, 6% w / w, 6.1% w / w, 6.2% w / w, 6.3% w / w, 6.4% w / w, or 6.5% w / w).

[0091] The resin may be pre-flushed before the isoprenoid composition contacts the resin. In the pre-flushing step, the resin may be completely wetted by passing a solvent through the resin. For example, the resin may be pre-flushed by passing 1 bed volume (BV) to 4 BV (e.g., 1 BV, 2 BV, 3 BV, or 4 BV) of solvent through the resin. In some embodiments, the pre-flushing step comprises passing 2 BV of solvent through the resin. The pre-flushing step may be performed by passing the solvent through the resin so that the minimum residence time in the column is at least 10 minutes. In the preliminary liquid-passing step, the solvent may be passed through the resin at a maximum downward superficial velocity of 0.5 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, or 1 cm / min to 3 cm / min). For example, in the preliminary liquid-passing step, the solvent may be passed through the resin at a maximum downward superficial velocity of 1 cm / min to 4 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 2 cm / min to 4 cm / min, 3 cm / min to 4 cm / min, or 2 cm / min to 3 cm / min). In some embodiments, the solvent is passed through the resin at a maximum upflow superficial velocity of 0.05 cm / min to 5 cm / min (e.g., 0.5 cm / min to 5 cm / min, 1 cm / min to 5 cm / min, 2 cm / min to 5 cm / min, 3 cm / min to 5 cm / min, 4 cm / min to 5 cm / min, 0.5 cm / min to 4 cm / min, 0.5 cm / min to 2 cm / min, 1 cm / min to 3 cm / min, 0.05 cm / min to 3 cm / min, 0.05 cm / min to 1 cm / min, or 0.05 cm / min to 0.1 cm / min). For example, the solvent may be passed through the resin at a maximum upward superficial velocity of 0.1 cm / min to 3 cm / min (e.g., 1 cm / min to 3 cm / min, 1 cm / min to 2 cm / min, 0.1 cm / min to 2 cm / min, 0.1 cm / min to 1 cm / min, or 2 cm / min to 3 cm / min). In the preliminary passing step, the farnesene composition may be passed through the resin to completely wet the resin.

[0092] After the pre-passing step, the farnesene composition may be exposed to the resin. In some embodiments, the resin is not subjected to a pre-passing step before the farnesene composition is exposed to the resin. The farnesene composition may be exposed to the resin for a minimum residence time on the resin of at least 10 minutes. The farnesene composition may be exposed to the resin at a maximum downward superficial velocity of 1 cm / min to 15 cm / min (e.g., 1 cm / min to 10 cm / min, 1 cm / min to 5 cm / min, 1 cm / min to 3 cm / min, 3 cm / min to 15 cm / min, 5 cm / min to 15 cm / min, 10 cm / min to 15 cm / min, or 5 cm / min to 10 cm / min). For example, the farnesene composition can be exposed to the resin at a maximum downward flow superficial velocity of 5 cm / min to 11 cm / min (e.g., 5 cm / min, 6 cm / min, 7 cm / min, 8 cm / min, 9 cm / min, 10 cm / min, or 11 cm / min). The farnesene composition can be exposed to the resin at a maximum upward flow superficial velocity of 0.1 cm / min to 10 cm / min (e.g., 0.1 cm / min to 8 cm / min, 0.1 cm / min to 6 cm / min, 0.1 cm / min to 4 cm / min, 0.1 cm / min to 1 cm / min, 1 cm / min to 10 cm / min, 3 cm / min to 10 cm / min, 5 cm / min to 10 cm / min, 8 cm / min to 10 cm / min, 4 cm / min to 8 cm / min, or 2 cm / min to 6 cm / min). For example, the farnesene composition can be exposed to the resin at a maximum upflow superficial velocity of 1 cm / min to 5 cm / min (e.g., 1 cm / min, 2 cm / min, 3 cm / min, 4 cm / min, or 5 cm / min).

[0093] The chromatography step may include regenerating the resin. Regenerating the resin may include adding a polar solvent to the resin, removing the polar solvent from the resin, heating the resin, and cooling the resin. The regeneration step may include adding a polar solvent to the resin in an amount of 1 BV to 10 BV (e.g., 1 BV, 2 BV, 3 BV, 4 BV, 5 BV, 6 BV, 7 BV, 8 BV, 9 BV, or 10 BV). For example, the polar solvent is added to the resin in an amount of 2 BV to 6 BV (e.g., 2 BV, 3 BV, 4 BV, 5 BV, or 6 BV). In some embodiments, the polar solvent is added to the resin in an amount of about 4 BV. In the regeneration step, the resin may be heated. The resin can be heated to a temperature of 100°C to 200°C (e.g., 100°C to 180°C, 100°C to 160°C, 100°C to 140°C, 100°C to 120°C, 120°C to 200°C, 150°C to 200°C, 170°C to 200°C, 140°C to 180°C, or 120°C to 160°C). For example, the resin can be heated to a temperature of about 150°C. After heating the resin, it can be cooled to room temperature. The resin can be regenerated with any polar solvent. For example, the polar solvent can be methanol or isopropanol.

[0094] The chromatography step may be carried out under N2.

[0095] Purity of isolated isoprenoids Disclosed herein is a composition containing a highly purified isoprenoid recovered from an isoprenoid composition, wherein the concentration of the isoprenoid relative to the total amount of the isoprenoid and one or more impurities contained in the composition can be, for example, 95% (w / w) to 100% (w / w) or more, such as 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more. In some embodiments, the isoprenoid is present in an amount of 98% (w / w) to 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98. 7%(w / w), 98.8%(w / w), 98.9%(w / w), 99%(w / w), 99.1%(w / w), 99.2%(w / w), 99.3%(w / w), 99.4%(w / w), 99.5%(w / w), 99.6%(w / w), 99.7%(w / w), 99.8%(w / w), 99.9%(w / w), or 100%(w / w)). In some embodiments, the isoprenoid is present in the composition in an amount of 99% (w / w) to 100% (w / w) of the total amount of the isoprenoid and the one or more impurities (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). For example, the concentration of the isoprenoid relative to the total amount of the isoprenoid and the one or more impurities contained in the composition can be about 99.5% (w / w) to about 100% (w / w) or more, for example, about 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w).

[0096] In some embodiments, the isoprenoid concentration relative to the total amount of the isoprenoid and the one or more impurities can be up to 100% (w / w), e.g., up to 100% (w / w), 99% (w / w), 98% (w / w), 97% (w / w), 96% (w / w), 95% (w / w), 94% (w / w), 93% (w / w), 92% (w / w), 91% (w / w), or 90% (w / w). In some embodiments, the isoprenoid concentration relative to the total amount of the isoprenoid and the one or more impurities can be greater than 95% (w / w), e.g., greater than 95% (w / w), greater than 96% (w / w), greater than 97% (w / w), greater than 98% (w / w), greater than 99% (w / w), greater than 99.1% (w / w), greater than 99.2% (w / w), greater than 99.3% (w / w), greater than 99.4% (w / w), greater than 99.5% (w / w), greater than 99.6% (w / w), greater than 99.7% (w / w), greater than 99.8% (w / w), or greater than 99.9% (w / w).

[0097] In some embodiments, an isoprenoid-containing composition of the present disclosure can include an isoprenoid and one or more impurities such that the concentration of the one or more impurities relative to the total amount of the isoprenoid and the one or more impurities is about 0.1% (w / w) to about 0.5% (w / w), e.g., about 0.1% (w / w), 0.2% (w / w), 0.3% (w / w), 0.4% (w / w), or 0.5% (w / w). In some embodiments, the concentration of the one or more impurities relative to the total amount of the isoprenoid and the one or more impurities is less than 0.5% (w / w), e.g., less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), or less than 0.1% (w / w). In some embodiments, the concentration of the one or more impurities relative to the total amount of the isoprenoid and the one or more impurities is about 0.1% (w / w), about 0.2% (w / w), about 0.3% (w / w), about 0.4% (w / w), or about 0.5% (w / w).

[0098] Also disclosed herein are compositions, specifically comprising a high-purity composition of isoprenoid farnesene recovered from a farnesene composition, wherein the concentration of the farnesene relative to the total amount of the farnesene and one or more impurities in the composition can be, for example, 95% (w / w) to 100% (w / w) or more, such as 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w) or more. In some embodiments, the farnesene is present in an amount of 98% (w / w) to 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98.8% (w / w), 98.9% (w / w), 98.10% (w / w), 98.11% (w / w), 98.12% (w / w), 98.13% (w / w), 98.14% (w / w), 98.15% (w / w), 98.16% (w / w), 98.17% (w / w), 98.18% (w / w), 98.19% (w / w), 98.20% (w / w), 98.21% (w / w), 98.22% (w / w), 98.23% (w / w), 98.24% (w / w), 98.25% (w / w), 98.26% (w / w), 98.27% (w / w), 98.28% (w / w), 98.29% (w / w), 98.29% (w / w), 98.29% (w / w), 98.29% (w / w), 98.26% (w / w), 98.29 ... %(w / w), 98.8%(w / w), 98.9%(w / w), 99%(w / w), 99.1%(w / w), 99.2%(w / w), 99.3%(w / w), 99.4%(w / w), 99.5%(w / w), 99.6%(w / w), 99.7%(w / w), 99.8%(w / w), 99.9%(w / w), or 100%(w / w)). In some embodiments, the farnesene is present in the composition in an amount of 99% (w / w) to 100% (w / w) of the total amount of the farnesene and the one or more impurities (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). For example, the concentration of the farnesene relative to the total amount of the farnesene and the one or more impurities contained in the composition can be about 99.5% (w / w) to about 100% (w / w) or more, e.g., about 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w).

[0099] The isolated isoprenoids of the present disclosure can be purified from the isoprenoid composition to a purity of about 95% (w / w) to about 100% (w / w) (e.g., 95% (w / w) to 100% (w / w) or more, e.g., 95% (w / w), 95.5% (w / w), 96% (w / w), 96.5% (w / w), 97% (w / w), 97.5% (w / w), 98% (w / w), 98.5% (w / w), 99% (w / w), 99.5% (w / w), or 100% (w / w), or more). The isoprenoid can be extracted from the isoprenoid composition at a purity of about 98% (w / w) to about 100% (w / w) (e.g., 98% (w / w), 98.1% (w / w), 98.2% (w / w), 98.3% (w / w), 98.4% (w / w), 98.5% (w / w), 98.6% (w / w), 98.7% (w / w), 98.8% (w / w)). ), 98.9% (w / w), 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). For example, the isoprenoid is purified from the isoprenoid composition to a purity of about 99% (w / w) to about 100% (w / w) (e.g., 99% (w / w), 99.1% (w / w), 99.2% (w / w), 99.3% (w / w), 99.4% (w / w), 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w)). In some embodiments, the isoprenoid is purified from the isoprenoid composition to a purity of about 99.5% (w / w) to about 100% (w / w) or greater, e.g., about 99.5% (w / w), 99.6% (w / w), 99.7% (w / w), 99.8% (w / w), 99.9% (w / w), or 100% (w / w).

[0100] In some embodiments, the concentration of the farnesene relative to the combined amount of the farnesene and the one or more impurities may be up to 100% (w / w), e.g., up to 100% (w / w), 99% (w / w), 98% (w / w), 97% (w / w), 96% (w / w), 95% (w / w), 94% (w / w), 93% (w / w), 92% (w / w), 91% (w / w), or 90% (w / w). In some embodiments, the concentration of the farnesene relative to the combined amount of the farnesene and the one or more impurities may be greater than 95% (w / w), e.g., greater than 95% (w / w), greater than 96% (w / w), greater than 97% (w / w), greater than 98% (w / w), greater than 99% (w / w), greater than 99.1% (w / w), greater than 99.2% (w / w), greater than 99.3% (w / w), greater than 99.4% (w / w), greater than 99.5% (w / w), greater than 99.6% (w / w), greater than 99.7% (w / w), greater than 99.8% (w / w), or greater than 99.9% (w / w).

[0101] In some embodiments, the farnesene-containing compositions of the present disclosure may include farnesene and one or more impurities such that the concentration of the one or more impurities relative to the total amount of farnesene is from about 0.1% (w / w) to about 0.5% (w / w), e.g., about 0.1% (w / w), 0.2% (w / w), 0.3% (w / w), 0.4% (w / w), or 0.5% (w / w). In some embodiments, the concentration of the one or more impurities relative to the total amount of farnesene and the one or more impurities is less than 0.5% (w / w), e.g., less than 0.4% (w / w), less than 0.3% (w / w), less than 0.2% (w / w), or less than 0.1% (w / w). In some embodiments, the concentration of the one or more impurities relative to the total amount of the farnesene and the one or more impurities is about 0.1% (w / w), e.g., about 0.2% (w / w), about 0.3% (w / w), about 0.4% (w / w), or about 0.5% (w / w).

[0102] The impurities in the isolated composition containing the high-purity isoprenoid may include, for example, polar impurities. In some embodiments, the composition contains a high-purity isoprenoid and one or more impurities. The one or more impurities may include, for example, farnesol and farnesene epoxide.

[0103] The composition may contain impurities, including farnesol, which may have a concentration of 0 mg / mL to 1 g / L (e.g., 0.2 g / L to 1 g / L, 0.4 g / L to 1 g / L, 0.6 g / L, 0.6 g / L to 1 g / L, 0.8 g / L to 1 g / L, 0 mg / mL to 0.8 g / L, 0 mg / mL to 0.6 g / L, 0 mg / mL to 0.4 g / L, 0 mg / mL to 0.2 g / L, 0.4 g / L to 0.6 g / L, or 0.2 g / L to 0.8 g / L). For example, the farnesol concentration can be 0 mg / L to 600 mg / L (e.g., 0 mg / mL to 500 mg / mL, 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 600 mg / mL, 200 mg / mL to 600 mg / mL, 300 mg / mL to 600 mg / mL, 400 mg / mL to 600 mg / mL, 500 mg / mL to 600 mg / mL, 200 mg / mL to 400 mg / mL, or 300 mg / mL to 500 mg / mL). In some embodiments, the farnesol concentration in the composition comprising the high-purity isoprenoid is less than 500 mg / L. The composition may contain one or more impurities, including the isoprenoid and farnesene epoxide. The farnesene epoxide may be present in the composition at a concentration of 0 mg / mL to 500 mg / L (e.g., 0 mg / mL to 400 mg / mL, 0 mg / mL to 300 mg / mL, 0 mg / mL to 200 mg / mL, 0 mg / mL to 100 mg / mL, 100 mg / mL to 500 mg / mL, 200 mg / mL to 500 mg / mL, 300 mg / mL to 500 mg / mL, 400 mg / mL to 500 mg / mL, 200 mg / mL to 400 mg / mL, or 100 mg / mL to 300 mg / mL). The composition can include the isoprenoid and a farnesene epoxide impurity having a concentration of 0 mg / L to 200 mg / L (e.g., 0 mg / L to 150 mg / L, 0 mg / L to 100 mg / L, 0 mg / L to 50 mg / L, 50 mg / L to 200 mg / L, 100 mg / L to 200 mg / L, 150 mg / L to 200 mg / L, 20 mg / L to 100 mg / L, or 50 mg / L to 150 mg / L). For example, the concentration of the farnesene epoxide can be less than 150 mg / L.

[0104] Cultivation and fermentation methods Materials and methods for the maintenance and growth of microbial cultures are well known to those skilled in the art of microbiology or fermentation science (see, e.g., Bailey et al., Biochemical Engineering Fundamentals, second edition, McGraw Hill, New York, 1986). The requirements for appropriate media, pH, temperature, and aerobic, microaerobic, or anaerobic conditions must be considered depending on the specific requirements of the host cell, fermentation, and process.

[0105] In some embodiments, the isoprenoids described herein can be isolated from host cells capable of producing isoprenoids. The host cell culture can be carried out in a suitable medium in a suitable container, including, but not limited to, a cell culture plate, a flask, or a fermenter. Furthermore, the method can be carried out at any fermentation scale known in the art to support the industrial production of microbial products. Any suitable fermenter can be used, including a stirred tank fermenter, an airlift fermenter, a bubble fermenter, or any combination thereof. In certain embodiments utilizing Saccharomyces cerevisiae as the host cell, the strain can be grown in a fermenter as described in detail by Kosaric, et al., in Ullmann's Encyclopedia of Industrial Chemistry, Sixth Edition, Volume 12, pages 398-473, Wiley-VCH Verlag GmbH & Co. KDaA, Weinheim, Germany.

[0106] In some embodiments, the medium is any medium capable of supporting the growth and viability of a microorganism capable of producing a heterologous product. In some embodiments, the medium is an aqueous medium containing assimilable carbon, nitrogen, and phosphate sources. Such a medium may also contain appropriate salts, minerals, metals, and other nutrients. In some embodiments, the carbon source and each of the essential cellular nutrients are gradually or continuously added to the fermentation medium, and each required nutrient is maintained at the minimum level essentially required for efficient assimilation by the growing cells, for example, according to a predetermined cell growth curve based on the metabolic or respiratory function of the cells to convert the carbon source into biomass.

[0107] Suitable conditions and suitable media for culturing microorganisms are well known in the art. In some embodiments, the suitable media is supplemented with one or more additional agents, such as an inducer (e.g., when one or more nucleotide sequences encoding a gene product are under the control of an inducible promoter), a repressor (e.g., when one or more nucleotide sequences encoding a gene product are under the control of a repressible promoter), or a selection agent (e.g., an antibiotic for selecting for microorganisms containing genetic modifications).

[0108] host cell line In some embodiments of the present disclosure, the host cell is a yeast cell. Yeast cells useful in conjunction with the compositions and methods described herein include yeast cells deposited with a microbial depository (e.g., IFO, ATCC, etc.), such as, among others, Aciculoconidium, Ambrosiozyma, Arthroascus, Arxiozyma, Ashbya, Babjevia, Bensingtonia, Botryoascus, Botryozyma, Brettanomyces, Bullera, Bulleromyces, Candida, Citeromyces, Clavispora, Cryptococcus, Cystofilobasidium, Debaryomyces, Dekkara, Dipodascopsis, Dipodascus, Eeniella, Endomycopsella, Eremascus, Eremothecium, Erythrobasidium, Fellomyces, Filobasidium, Galactomyces, Geotrichum, Guilliermondella, Hanseniaspora, Hansenula, Hasegawaea, Homo sapiens ... ltermannia, Hormoascus, Hyphochia, Issatchenkia, Kloeckera, Kloeckeraspora, Kluyveromyces, Kondoa, Kuraishia, Kurtzmanomyces, Le ucosporidium, Lipomyces, Lodderomyces, Malassezia, Metschnikowia, Mrakia, Myxozyma, Nadsonia, Nakazawaea, Nematospora, Ogataea, Oosp oridium, Pachysolen, Phachytichospora, Phaffia, Pichia, Rhodosporidium, Rhodotorula, Saccharomyces, Saccharomycodes, Saccharomycopsis, Saitoella, Sakaguchia, Saturnospora, Schizoblastosporion, chizosaccharomyces, Schwanniomyces, Sporidiobolus, Sporobolomyces,Examples include those belonging to the genera Sporopachydermia, Stephanoascus, Sterigmatomyces, Sterigmatosporidium, Symbiotaphrina, Sympodiomyces, Sympodiomycopsis, Torulaspora, Trichosporiella, Trichosporon, Trigonopsis, Tsuchiyaea, Udeniomyces, Waltomyces, Wickerhamia, Wickerhamiella, Williopsis, Yamadazyma, Yarrowia, Zygoascus, Zygosaccharomyces, Zygowilliopsis, and Zygozyma.

[0109] In some embodiments, the strain is Saccharomyces cerevisiae, Pichia pastoris, Schizosaccharomyces pombe, Dekkera bruxellensis, Kluyveromyces lactis (formerly known as Saccharomyces lactis), Kluyveromyces marxianus, Arxula adeninivorans, or Hansenula polymorphs (now known as Pichia angusta). In some embodiments, the host microorganism is a strain of the Candida genus, such as Candida lipolytica, Candida guilliermondii, Candida krusei, Candida pseudotropicalis, or Candida utilis.

[0110] In some embodiments, the host cell is Saccharomyces cerevisiae. Saccharomyces cerevisiae strains suitable for culture to produce the isoprenoids disclosed herein include, but are not limited to, baker's yeast, CBS7959, CBS7960, CBS7961, CBS7962, CBS7963, CBS7964, IZ-1904, TA, BG-1, CR-1, SA-1, M-26, Y-904, PE-2, PE-5, VR-1, BR-1, BR-2, ME-2, VR-2, MA-3, MA-4, CAT-1, CB-1, NR-1, BT-1, CEN.PK, CEN.PK2, and AL-1. In some embodiments, the host cell is a strain of Saccharomyces cerevisiae selected from the group consisting of PE-2, CAT-1, VR-1, BG-1, CR-1, and SA-1. In certain aspects, the Saccharomyces cerevisiae strain is PE-2. In certain embodiments, the Saccharomyces cerevisiae strain is CAT-1. In some aspects, the Saccharomyces cerevisiae strain is BG-1.

[0111] In some embodiments, the host cell is Kluyveromyces marxianus. Kluyveromyces marxianus may offer several advantages for industrial production, including high temperature tolerance, acid tolerance, natural lactose uptake, and fast growth rate. Advantageously, this yeast has sufficient genetic similarity with Saccharomyces cerevisiae to allow the use of similar or identical promoters and codon-optimized genes between the two yeast species. [Example]

[0112] The following examples are presented to provide one of ordinary skill in the art with a description of how the compositions and methods described herein can be used, made, and evaluated, and are intended to be merely exemplary of the invention and are not intended to limit the scope of what the inventors regard as their invention.

[0113] Example 1. Isolation of High-Purity Farnesene from Distilled Farnesene C 15 The hydrocarbon farnesene was produced using fermentation and isolated using centrifugation and distillation. The distilled farnesene was typically 97% (w / w) pure and contained approximately 1% oxygenate impurities, which needed to be removed to ppm levels before being used as a monomer in polymerization reactions. An adsorption-based purification process was used to purify the distilled farnesene to a purity of greater than 99% w / w. The process used to purify the farnesene included an adsorption step through a basic alumina bed, followed by draining the bed with pressurized nitrogen gas, in situ regeneration using methanol solvent, and in situ activation of the adsorbent. These experiments were performed in a 15 cm long, 2.5 cm diameter glass column plumbed through a skid using a dual-head pump, air valve system, and variable nitrogen flow.

[0114] The adsorbent used was ESM-20 (28x48 mesh) basic alumina manufactured by ASM Catalysts, LLC. The adsorption characteristics are shown in Tables 1 and 2. The Langmuir parameters for ESM-20 basic alumina were calculated using equilibrium data for the pure components obtained in the laboratory at 22°C. [Table 1] [Table 2]

[0115] The specifications for the alumina treated farnesene product are shown in Table 3. [Table 3]

[0116] Feed Impurity Profile Table 4 shows the variability of the feed impurity profiles used as starting materials to isolate high purity farnesene. [Table 4]

[0117] Process Steps Adsorbent filling The column was packed with alumina and the mass of the alumina packing was recorded. The device was pressure tested to 50 psig using dry nitrogen.

[0118] Preliminary liquid flow Valves / equipment were reconfigured as necessary and pumping of distilled farnesene was initiated. The pump flow rate was set and distilled farnesene flow was initiated. 2BV of material was run through the column and collected as product. Samples from this process were checked for farnesol and epoxide concentrations and then stabilized with antioxidants.

[0119] The adsorption process was operated at ambient temperature without active heating or cooling. This flow-through step was mildly exothermic, and a maximum temperature rise of approximately 7°C was observed in this laboratory-scale column, based on the surface temperature recorded on a 2.5 x 15 cm stainless steel column. This initial exotherm subsided after the first bed volume of feed was processed, and the column was completely wetted. Consistent with laboratory experience, a maximum transient temperature rise of approximately 8°C was recorded at the 300 L scale, based on the temperature difference between the process inlet and outlet lines. No temperature measurements were made within the alumina bed, either in the laboratory or at the pilot scale.

[0120] For both the pre-fill and adsorption steps, the maximum superficial velocity and corresponding mass flow rate were determined by the minimum residence time requirement, the bed fluidization limit (applicable for upflow operation), and the particle attrition limit (applicable for downflow operation). In particular, the feed should be processed more slowly during the first bed volume (pre-fill), when the column is not yet filled with liquid. The following design limits were observed during operation of this process: [Table 5]

[0121] adsorption The pump flow rate was set and the injection of distilled farnesene was initiated. Samples after each BV of this process were checked for farnesol and epoxide concentrations. This material was recovered as product and stabilized with antioxidants.

[0122] Recovery of farnesene holdup The equipment was reconfigured as needed to switch to dry nitrogen gas. The dry nitrogen gas was set to a specified flow rate. The nitrogen was allowed to flow for a period of time to drain the liquid. The drained material was collected in a container separate from the product. Samples of the process were collected after each BV for farnesol and epoxide concentrations. This material was collected as product and stabilized with antioxidants.

[0123] Hold-up recovery (drainage) To maximize yield, farnesene retained in the column's void space was recovered by draining the column while blowing dry nitrogen through it. The recovered farnesene was sometimes blended into the product if it was close enough to specification, or sometimes recycled back to the feed tank for further reprocessing. The total bed porosity was approximately 86%. The expected percentage of recovered holdup was 43–48%, based on the interparticle void space in the bed. This was consistent with what was observed in the laboratory. Typically, a 15 cm laboratory column yielded within-specification product in 5–7 bed volumes, depending on the impurity concentration profile of the feed. Combined with 40% holdup recovery, this represents an overall process yield of 92%. The number of bed volumes of product obtained, and the corresponding yield, were expected to be even higher in a larger column with improved bed utilization compared to the laboratory-scale column.

[0124] Alumina Regeneration The alumina bed was regenerated. Valves / apparatus were reconfigured as necessary, and methanol pumping was initiated. The pump was set to the specified flow rate, and methanol flow was initiated. A specified amount of material was passed through the column. Because farnesene is nonvolatile and reacts to form significant amounts of dimers and polymers within minutes at fairly low temperatures (<100°C), traditional temperature and pressure swing adsorption cycles were not suitable for this application. Instead, a hybrid chemical swing / temperature swing adsorption approach was used to regenerate the alumina bed. Methanol was used as the solvent of choice due to its low cost, high volatility, and high affinity for alumina. The C15 impurity was displaced by flowing methanol countercurrently with the farnesene through the bed (methanol downflow) at ambient temperature. The effluent methanol was captured and saved for later purification. The bed was then drained using pressurized nitrogen to remove any readily accessible liquid holdup (typically 50-60% of the total MeOH holdup). The methanol remaining in the bed after draining came from heating the bed in situ to temperatures above 150°C. This methanol vapor effluent was condensed and combined with the liquid effluent from the previous step. Finally, the bed was cooled back to ambient temperature in preparation for the next adsorption cycle.

[0125] Bed regeneration has been demonstrated at laboratory scale through five regeneration passes. Four bed volumes of methanol, consisting of >90% regeneration solvent per pass, were used to regenerate the bed for each pass.

[0126] Solvent Recycling The waste methanol effluent from the bed regeneration step was purified and recycled. In the laboratory, methanol was purified using a single-step evaporation at 43°C and 150 torr vacuum. These conditions were selected to achieve >90% methanol recovery per pass, with a distilled methanol purity of 99.9%, while keeping the evaporator temperature as low as possible to reduce the possibility of polymer formation and fouling within the evaporator.

[0127] Alternative evaporator conditions of 350 torr and 60°C were estimated using process modeling tools. These conditions should result in distilled methanol with comparable yield and purity, allowing operation using a condenser temperature of 35°C.

[0128] In situ activation The apparatus was reconfigured as necessary to move the nitrogen flow through the condenser system. The cooling water was set at the desired flow rate and temperature to ensure the condenser achieved the desired temperature. The dry nitrogen gas was set at the desired flow rate. A steam flow was passed through a jacket wrapped around the column at the desired flow rate and temperature. The column temperature was monitored to ensure it reached the desired temperature. The condensed material was collected in a container separate from the product.

[0129] Experimental results Laboratory-scale purification of the distilled farnesene feed was completed using ESM-20 (28x48 mesh) basic alumina. The glass laboratory column was 2.5 cm ID x 15 cm. The laboratory column did not incorporate a distributor and was run in an upflow fashion to ensure uniform radial distribution of the feed throughout the alumina bed. The operating conditions used in the laboratory are summarized in Table 6 below. [Table 6]

[0130] Six adsorption passes and five regeneration passes were completed consecutively using one lot of farnesene on a single alumina packed bed. Four bed volumes of virgin methanol were used for the first bed regeneration, and recycled methanol was used for subsequent regeneration passes. 90-93% of the waste methanol was recovered in each evaporator pass, and GC analysis of the methanol distillate confirmed that its C15 content was less than 0.1%. The capacity loss observed after the first regeneration (see Table 7) closely resembles previous experimental data and is likely due to bed spallation / wear caused by the introduction of approximately 10 psi of nitrogen during the drainage process. After the first regeneration, the bed capacity remained essentially constant to within the limits of experimental error. The laboratory experiment was terminated after a net recycle rate of 75% was demonstrated. Based on the data obtained, it should be possible to continue bed regeneration for well beyond five regeneration passes. Across multiple experiments, typical high-purity farnesene contained the components shown in Table 7 and Figure 1. [Table 7]

[0131] Typical impurity profile of the product A typical impurity profile for the chromatographic farnesene product is shown in the table below and in Figure 1. [Table 8]

[0132] Example 2. Purification of farnesene using various resins Chromatography was used to purify the farnesene composition using a variety of resin types.

[0133] Alumina Resin Farnesene was purified using (1) basic alumina resin from Sorbent Technology, (2) basic alumina resin from Sigma Aldrich, or (3) Activity Super 1 activated alumina from Sorbent Technology in the same manner as described in Example 1. For alumina stripping performed using basic alumina from Sorbent Technology (FIG. 2A), basic alumina from Sigma Aldrich (FIG. 2B), and Activity Super I activated alumina from Sorbent Technology (FIG. 2C), the purity of the resulting farnesene was characterized and the concentrations of various impurities, including farnesene epoxide, farnesol, and farneseic acid, were measured. These measurements are summarized in Tables 9-12 below. [Table 9] [Table 10] [Table 11]

[0134] Alumina and Silica Resin The purity of farnesene was characterized by comparison with silica gel Merck resin after chromatography using Sigma-Aldrich aluminum oxide resin. These experiments were performed on dry-packed columns (1.5 cm x 11.2 cm, 19.8 mL) in the same manner as described in Example 1. After filtration of crude farnesene through basic alumina (9 bed volumes), farnesol (0.1%) broke through, and no acidic species (e.g., farneseic acid) were detected. Filtration of crude farnesene through silica gel resulted in breakthrough of farnesol and farneseic acid after 5 CV, with other acidic species observed after 4 CV. At a 1.5 L scale, after filtration of crude farnesene through basic alumina (7 bed volumes), farnesol broke through, and no acidic species (e.g., farneseic acid) were detected.

[0135] Example 3. Alumina Treatment of Cadinane to Remove Polar Impurities Alumina treatment of sandalwood-spiked cadinane to remove santalol A column (1 cm ID x 14 cm, 11 mL BV) was dry-packed with Sorbent Technologies basic alumina, Activation I adsorbent. Cadinane (194.0 g) was mixed with sandalwood oil (15.4 g) containing santalene and santalol. The cadinane was purified using the same method as described in Example 1 above. Analysis of the mixture by GC-MS revealed an area purity of 1.2 area% santalol. The cadinane-sandalwood mixture was loaded onto the alumina column at a flow rate of 0.7–0.9 mL / min (3–4 BV / h). The eluate was collected in 1 BV aliquots (11 mL) and analyzed by GC-MS to determine the decrease in santalol levels.

[0136] The breakthrough data of santalol after alumina column treatment was determined, and as shown in Figure 3 and Table 12, the alumina column was able to reduce santalol from over 12000 ppm to below detection levels for the first three bed volumes. [Table 12]

[0137] Alumina treatment of cadinane spiked with hexahydrofarneseic acid (HHFA) A column (1 cm ID x 14 cm, 11 mL BV) was dry packed with Sorbent Technologies basic alumina, Activation I sorbent. This cadinane was purified using the same method as described in Example 1 above. Cadinane (198.5 g) was mixed with HHFA (13 g) to create a 211 g feed. This mixture was analyzed by GC-MS and found to have an area purity of 1.1 area % HHFA. The cadinane-HHFA mixture was loaded onto the alumina column at a flow rate of 0.7–0.9 mL / min (3–4 BV / h). The eluate was collected in 1 BV aliquots and analyzed by GC-MS to determine the reduction in HHFA levels.

[0138] As shown in Figure 4 and Table 13, no HHFA was observed in the first 2BV, demonstrating the utility of the alumina treatment to remove polar impurities present at above 11,000 ppm. [Table 13]

[0139] Example 4. Acid Treatment of Farnesene This experiment was conducted to determine whether acid treatment could successfully remove impurities from both distilled and crude farnesene to obtain a high purity farnesene product similar to that obtained from the methods described above in Examples 1-3. However, acid treatment of both crude and distilled farnesene presented a number of challenges, discussed below, which ultimately made purification of farnesene by acid treatment less effective than in the preceding examples.

[0140] To determine the feasibility of the acid treatment process for producing high-purity farnesene, a DOE (design of experiments) was conducted to evaluate the effects of temperature, time, and phosphoric acid loading. Measured responses were in terms of gas chromatographic (GC) area percent farnesene, farnesol, dimer, farnesene epoxide, total acid number (TAN), and color (Saybalt). The distilled farnesene used in these experiments contained approximately 8000 ppm farnesol and 250 ppm farnesene epoxide. Farnesene was typically added to a round-bottom flask, and the liquid was mixed at 340 rpm. Acid was then added, and the suspension was allowed to come to temperature. Aliquots of the acid-treated farnesene were taken at 30-minute intervals and centrifuged to remove the acid phase. tert-Butylcatechol (TBC, 100 ppm) was immediately added to the separated farnesene phase as a stabilizer to reduce oxidation.

[0141] Farnesol and farnesene epoxide were readily removed from farnesene by mixing with 2 wt% H3PO4 at 80-90 °C for 1.5-2 h (Figure 6A). Data describing the epoxide purification results were confounded by the presence of coeluting impurities during the reaction. Experiments in which acid-treated farnesene was spiked with farnesene epoxide showed that farnesene epoxide was removed from farnesene in less than 1 h. However, farnesene loss increased with increasing temperature due to the formation of farnesene dimers. This increase in dimers was accompanied by a decrease in the area purity of farnesene (Figure 6B). By reacting farnesene with acid at 80 °C for less than 3 h, the dimer content could be kept below 1 wt%.

[0142] The acid content of the treated farnesene was observed to be consistently low at high temperatures, regardless of the amount of acid used (Figure 7). The resulting farnesene had a pale yellow color compared to the colorless distilled farnesene. None of the factors examined showed a correlation with the color of the treated material. Several by-products were observed in the acid-treated farnesene. Liquid chromatography-mass spectrometry (LCMS) revealed that two of these compounds were cyclized C 15 Possible oxygenates and other impurities were shown to be alcohols, epoxides, and products of dehydration and isomerization of farnesene.

[0143] Using data from the experiments conducted, a predictive model was developed using response surface methodology (RSM). JMP Predictive Profiler was used to determine the most appropriate conditions for producing acid-treated farnesene meeting the specified specifications. The specified specifications included (1) a minimum area percent purity of 97 wt% farnesene, (2) a maximum farnesol content of 1000 ppm, and (3) a dimer level of 1% or less. Using these specified specifications, two scale-up experiments were conducted. The first scale-up test was conducted at an approximately 8 L scale at 80°C using 4 wt% H3PO4. The suspension was mixed for 2.5 hours and then cooled to room temperature. The farnesene was decanted, and a portion was stabilized with TBC and submitted for analysis. The overall mass yield recovery of this crude acid-treated farnesene was 99.3 wt%. The material produced met or exceeded the desired specifications, demonstrating that the model used to determine the appropriate conditions for meeting the desired specifications was highly predictive (Table 14). The resulting material was found to be pale yellow in color and had a TAN of 0.095 mg KOH / g farnesene. There was a desire to develop a second scale-up run of acid-treated material with a lower TAN and less color for testing of the adsorption layer. [Table 14]

[0144] The model was updated with color data and used to predict optimal conditions for scale-up. Distilled farnesene was treated with 6.9 wt% H3PO4 at 77 °C for 3 hours. The resulting suspension was cooled to room temperature, and the farnesene layer was collected and washed with water to remove acidic species. The washed organic layer was mixed with dilute NaOH to neutralize any remaining acidic components and then washed again with water. The farnesene was then dried over anhydrous magnesium sulfate. The overall mass yield of the purified farnesene was 98.3 wt%. The TAN and water content of this farnesene were significantly lower than those of the crude material. However, a pale yellow color was still evident. The coloring components could be removed using adsorbents such as activated carbon and clay. Therefore, a process envisioned for producing high-purity material from distilled farnesene would require one or more additional purification steps to address both color and acidity.

[0145] Acid treatment of crude farnesene was considered an attractive option to reduce overall purification costs because it could remove oxygenates and heavy components such as glycerides and surfactants in a single step, thus eliminating the need for a distillation step. To validate this approach, an experimental design was conducted using crude farnesene. As described above, the custom-designed approach was performed in a similar manner to that used with distilled farnesene. The factors evaluated were time, temperature, and H3PO4 loading. Crude farnesene was mixed at room temperature, H3PO4 was added, and the suspension was allowed to come to temperature. Aliquots were taken at 0.5, 1, 2, and 4 hours. The aliquots were centrifuged, and the farnesene layer was collected and tested.

[0146] Farnesol and farnesene epoxide were effectively removed from crude farnesene by mixing with 6 wt% H3PO4 at room temperature. Using less acid resulted in a monolayer of farnesene and the formation of a viscous brown gum ("acid phase") on the walls of the reaction vessel. A minimum of 4 wt% acid was required to prevent this brown gum from forming and keep the acid phase completely suspended. While dimer formation was negligible at room temperature, significant dimer formation was observed at higher temperatures. Concurrently, a decrease in the area percent of farnesene was observed (Figure 8A). The weight percent assay of selected acid-treated crude farnesene samples was below the 97 wt% specification. A rapid decrease in the weight percent assay of farnesene was observed with increasing mixing time (Table 15). Isomerization of farnesene and other sesquiterpenes increased significantly at 57 and 90 °C (Figure 8B). Farnesene isomerization was the primary cause of the low farnesene purity observed when assayed at all temperatures. Note that the low farnesene purity (20 wt%) at 4 hours at 90°C was due to temperature excursion. [Table 15]

[0147] Several attempts were made to reduce the acidity and color of the acid-treated crude farnesene by washing with water and base. These attempts resulted in the formation of a milky white emulsified farnesene layer. The emulsion remained intact even after centrifugation at 5000 rpm. When this emulsified farnesene was mixed with 8 wt% clay, a very pale yellow, transparent farnesene phase was obtained. As a result, the farnesene loss (estimated 8 wt%) associated with clay treatment rendered the acid treatment process incompatible with the process for farnesene purification described herein. The formation of isomerization products in the oxygenates during acid treatment made the process described in this example less attractive for removing oxygenates from farnesene.

[0148] conclusion Farnesol and farnesene epoxide can be easily removed from crude farnesene at much lower temperatures compared to distilled farnesene. However, crude farnesene tends to isomerize during the process, resulting in lower farnesene purity as measured by wt% assayed farnesene. Furthermore, purifying crude farnesene resulted in the formation of emulsions, which required additional process steps. Finally, several by-products were formed during the acid treatment of both crude and distilled farnesene, making purification of farnesene by acid treatment undesirable.

[0149] Other embodiments All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0150] While the invention has been described in connection with particular embodiments thereof, it will be understood that further modifications are possible, and that this application is intended to cover generally any variations, uses, or adaptations of the invention in accordance with the principles of the invention and to include departures from the invention which have become known or customarily practiced in the art to which the invention pertains, as applicable to the essential features described above, and in accordance with the scope of the appended claims.

[0151] Other embodiments are within the scope of the following claims.

[0152] Numbered Embodiments The invention disclosed herein is also represented by the following non-limiting recited paragraphs.

[0153] [1] A method for purifying farnesene, comprising: (a) providing a farnesene composition; and (b) purifying the farnesene from the farnesene composition of (a) by chromatography. The method comprising:

[0154] [2] The method of paragraph [1], wherein the farnesene composition comprises farnesene and one or more impurities.

[0155] [3] The method of paragraph [2], wherein the one or more impurities comprise one or more polar impurities, and optionally, the one or more polar impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, hexahydrofarneseic acid (HHFA), santalene, or santalol.

[0156] [4] The method of paragraph [2] or [3], wherein the farnesene composition has a concentration of one or more impurities of 1 ppm to 15,000 ppm.

[0157] [5] The method of paragraph [4], wherein the farnesene composition has a concentration of one or more impurities of 1000 ppm to 12,000 ppm.

[0158] [6] The method of paragraph [4], wherein the farnesene composition has a concentration of one or more impurities of 100 ppm to 1,000 ppm.

[0159] [7] The method of any one of paragraphs [1] to [6], wherein 4-tert-butylcatechol is added to the farnesene composition.

[0160] [8] The method of paragraph [7], wherein the 4-tert-butylcatechol is added to the farnesene composition to a concentration of 50 ppm to 150 ppm, and optionally, the 4-tert-butylcatechol is added to the farnesene composition to a concentration of about 100 ppm.

[0161] [9] The method of any one of paragraphs [1] to [8], wherein the farnesene composition is not treated with phosphoric acid prior to the purification step (b), and the farnesene composition is not treated with an acid prior to the purification step (b).

[0162]

[10] The method of paragraph [9], wherein the farnesene composition is not treated with an acid prior to the purification step (b).

[0163]

[11] The chromatography (a) Preliminary passage of the resin; (b) exposing the farnesene composition to the resin; and (c) recovering the farnesene from the resin.

[0164]

[12] The method of paragraph

[11] , wherein the resin comprises aluminum oxide.

[0165]

[13] The method of paragraph

[12] , wherein the aluminum oxide is basic aluminum oxide.

[0166]

[14] The method of paragraph

[12] , wherein the aluminum oxide is acidic aluminum oxide.

[0167]

[15] The method of paragraph

[12] , wherein the aluminum oxide is neutral aluminum oxide.

[0168]

[16] The method of paragraph

[11] , wherein the resin comprises silica.

[0169]

[17] The method of any one of paragraphs

[12] to

[16] , wherein the resin has a bulk density of 0.1 g / mL to 0.75 g / mL.

[0170]

[18] The method of paragraph

[17] , wherein the resin has a bulk density of 0.25 g / mL to 0.5 g / mL.

[0171]

[19] The method of any one of paragraphs

[11] to

[18] , wherein the resin has a pore volume of 0.1 mL / g to 1.5 mL / g.

[0172]

[20] The method of paragraph

[19] , wherein the resin has a pore volume of 0.5 mL / g to 1 mL / g.

[0173]

[21] The method of any one of paragraphs

[11] to

[20] , wherein the resin has a particle distribution size of 25 μm to 800 μm, and optionally, the resin has a particle distribution size of 100 μm to 800 μm.

[0174]

[22] The method of paragraph

[21] , wherein the resin has a particle distribution size of 300 μm to 600 μm.

[0175]

[23] The method of any one of paragraphs

[11] to

[22] , wherein the resin has an approximate moisture content of 0.01% w / w to 9% w / w.

[0176]

[24] The method of paragraph

[23] , wherein the resin has an approximate moisture content of 4.5% w / w to 6.5% w / w.

[0177]

[25] The method according to any one of paragraphs

[11] to

[24] , wherein the preliminary liquid passing step includes passing a solvent through the resin to completely wet the resin.

[0178]

[26] The method according to paragraph

[25] , wherein the preliminary passing step comprises passing the solvent through the resin in an amount of 1 bed volume (BV) to 4 BV.

[0179]

[27] The method of paragraph

[26] , wherein the preliminary flow step comprises flowing the solvent through the resin in an amount of 2BV.

[0180]

[28] The method according to any one of paragraphs

[25] to

[27] , wherein the solvent is passed through the resin in the preliminary passing step with a minimum residence time in the column of at least 10 minutes.

[0181]

[29] The method according to any one of paragraphs

[25] to

[28] , wherein the solvent is passed through the resin at a maximum downward superficial velocity of 0.5 cm / min to 5 cm / min in the preliminary passing step.

[0182]

[30] The method according to paragraph

[29] , wherein the solvent is passed through the resin at a maximum downward superficial velocity of 1 cm / min to 4 cm / min in the preliminary passing step.

[0183]

[31] The method of any one of paragraphs

[25] to

[30] , wherein the solvent is passed through the resin at a maximum upward superficial velocity of 0.05 cm / min to 5 cm / min.

[0184]

[32] The method of paragraph

[31] , wherein the solvent is passed through the resin at a maximum upward superficial velocity of 0.1 cm / min to 3 cm / min.

[0185]

[33] The method of any one of paragraphs

[11] to

[30] , wherein the farnesene composition is exposed to the resin after the pre-liquidation step.

[0186]

[34] The method of paragraph

[33] , wherein the farnesene composition is exposed to the resin for a minimum residence time on the resin of at least 10 minutes.

[0187]

[35] The method of paragraph

[33] or

[34] , wherein the farnesene composition is exposed to the resin at a maximum downward superficial velocity of 1 cm / min to 15 cm / min.

[0188]

[36] The method of paragraph

[35] , wherein the farnesene composition is exposed to the resin at a maximum downward superficial velocity of 5 cm / min to 11 cm / min.

[0189]

[37] The method of any one of paragraphs

[33] to

[36] , wherein the farnesene composition is exposed to the resin at a maximum upflow superficial velocity of 0.1 cm / min to 10 cm / min.

[0190]

[38] The method of paragraph

[37] , wherein the farnesene composition is exposed to the resin at a maximum upflow superficial velocity of 1 cm / min to 5 cm / min.

[0191]

[39] The method of any one of paragraphs

[11] to

[38] , wherein steps (a) to (c) are repeated 2 to 20 times.

[0192]

[40] The method of paragraph

[39] , wherein steps (a) to (c) are repeated 2 to 20 times.

[0193]

[41] The method of paragraph

[40] , wherein steps (a) to (c) are repeated 2 to 10 times.

[0194]

[42] The method of paragraph

[41] , wherein steps (a) to (c) are repeated six times.

[0195]

[43] The method of paragraph

[41] , wherein steps (a) to (c) are repeated 2 to 4 times.

[0196]

[44] The method of any one of paragraphs

[11] to

[43] , wherein the resin is regenerated after recovery of the farnesene from the resin in step (c).

[0197]

[45] The resin, (a) adding a polar solvent to the resin; (b) removing the polar solvent from the resin; (c) heating the resin; and (d) cooling the resin. The method according to paragraph

[44] , which is reproduced by

[0198]

[46] The method of paragraph

[45] , wherein step (a) is carried out by adding the polar solvent to the resin in an amount of 1 BV to 10 BV.

[0199]

[47] The method of paragraph

[46] , wherein the polar solvent is added to the resin in an amount of 2BV to 6BV.

[0200]

[48] ​​The method of paragraph

[47] , wherein the polar solvent is added to the resin in an amount of about 4 BV.

[0201]

[49] The method of any one of paragraphs

[45] to

[48] , wherein step (c) is carried out by heating the resin to a temperature of 100°C to 200°C.

[0202]

[50] The method of paragraph

[49] , wherein the resin is heated to a temperature of about 150° C.

[0203]

[51] The method according to any one of paragraphs

[45] to

[50] , wherein the cooling in step (d) is carried out by cooling the resin to room temperature.

[0204]

[52] The method according to any one of paragraphs [1] to

[51] , wherein the chromatography is carried out under N2.

[0205]

[53] The method of any one of paragraphs [1] to

[52] , wherein the farnesene is purified from the farnesene composition to a purity of about 95% (w / w) to about 100% (w / w).

[0206]

[54] The method of paragraph

[53] , wherein the farnesene is purified from the farnesene composition to a purity of about 98% (w / w) to about 100% (w / w).

[0207]

[55] The method of paragraph

[54] , wherein the farnesene is purified from the farnesene composition to a purity of about 99% (w / w) to about 100% (w / w).

[0208]

[56] The method of paragraph

[55] , wherein the farnesene composition is purified to a purity of about 99.5% (w / w) to about 100% (w / w).

[0209]

[57] A composition comprising farnesene, the composition being produced by the method of any one of paragraphs [1] to

[56] .

[0210]

[58] The composition of paragraph

[57] , wherein the farnesene has a purity of about 95% (w / w) to about 100% (w / w).

[0211]

[59] The composition of paragraph

[58] , wherein the farnesene has a purity of about 98% (w / w) to about 100% (w / w), and optionally, the farnesene has a purity of about 99% (w / w) to about 100% (w / w).

[0212]

[60] The composition of paragraph

[59] , wherein the farnesene has a purity of about 99.5% (w / w) to about 100% (w / w).

[0213]

[61] The composition of any one of paragraphs

[57] to

[60] , comprising one or more impurities including farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0214]

[62] A composition comprising farnesene and one or more impurities, wherein the farnesene has a purity of about 95% (w / w) to about 100% (w / w), and the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, a fatty acid, a sterol, HHFA, santalene, or santalol.

[0215]

[63] The composition of paragraph

[61] or

[62] , comprising farnesol and having a farnesol concentration of 0 mg / mL to 1 g / L.

[0216]

[64] The composition of paragraph

[63] , wherein the concentration of farnesol is 0 mg / L to 600 mg / L.

[0217]

[65] The composition of paragraph

[64] , wherein the concentration of farnesol is less than 500 mg / L.

[0218]

[66] The composition of any one of paragraphs

[61] to

[65] , comprising farnesene epoxide and having a farnesol concentration of 0 mg / mL to 500 mg / L.

[0219]

[67] The composition of paragraph

[66] , wherein the concentration of the farnesene epoxide is 0 mg / L to 200 mg / L.

[0220]

[68] The composition of paragraph

[67] , wherein the concentration of the farnesene epoxide is less than 150 mg / L.

[0221]

[69] A composition comprising farnesene and one or more carriers, diluents, or excipients, wherein the purity of the farnesene is from about 99.5% (w / w) to about 100% (w / w).

[0222]

[70] The composition of paragraph

[69] , wherein the farnesene is present with one or more impurities, and the one or more impurities are present at a concentration of about 0.5% (w / w) or less.

[0223]

[71] The composition of paragraph

[70] , wherein the one or more impurities are present at a concentration of about 0.4% (w / w) or less, optionally the one or more impurities are present at a concentration of about 0.3% (w / w) or less, optionally the one or more impurities are present at a concentration of about 0.2% (w / w) or less, and optionally the one or more impurities are present at a concentration of about 0.1% (w / w) or less.

[0224]

[72] The composition of paragraph

[70] or

[71] , wherein the one or more impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0225]

[73] A method for purifying an isoprenoid, comprising: (a) providing an isoprenoid composition, and (b) purifying the isoprenoid from the isoprenoid composition of (a) by chromatography. The method comprising:

[0226]

[74] The method of paragraph

[73] , wherein the isoprenoid composition comprises an isoprenoid and one or more impurities.

[0227]

[75] The method of paragraph

[74] , wherein the one or more impurities include one or more polar impurities.

[0228]

[76] The method of paragraph

[75] , wherein the one or more polar impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0229]

[77] The method of any one of paragraphs

[74] to

[76] , wherein the isoprenoid composition has a concentration of one or more impurities of 1 ppm to 15,000 ppm.

[0230]

[78] The method of paragraph

[77] , wherein the isoprenoid composition has a concentration of one or more impurities of 1000 ppm to 12,000 ppm.

[0231]

[79] The method of paragraph

[77] , wherein the isoprenoid composition has a concentration of one or more impurities of 100 ppm to 1,000 ppm.

[0232]

[80] The method of any one of paragraphs

[73] to

[79] , wherein 4-tert-butylcatechol is added to the isoprenoid composition.

[0233]

[81] The method of paragraph

[80] , wherein the 4-tert-butylcatechol is added to the isoprenoid composition to a concentration of 50 ppm to 150 ppm, and optionally, the 4-tert-butylcatechol is added to the isoprenoid composition to a concentration of about 100 ppm.

[0234]

[82] The method of any one of paragraphs

[73] to

[81] , wherein the isoprenoid composition is not treated with phosphoric acid prior to the purification step (b).

[0235]

[83] The method of paragraph

[82] , wherein the isoprenoid composition is not treated with an acid prior to the purification step (b).

[0236]

[84] The chromatography (a) Preliminary passage of the resin; (b) exposing the isoprenoid composition to the resin; and (c) recovering the isoprenoid from the resin. The method according to any one of paragraphs

[73] to

[83] , comprising:

[0237]

[85] The method of paragraph

[84] , wherein the resin comprises aluminum oxide.

[0238]

[86] The method of paragraph

[85] , wherein the aluminum oxide is basic aluminum oxide.

[0239]

[87] The method of paragraph

[85] , wherein the aluminum oxide is acidic aluminum oxide.

[0240]

[88] The method of paragraph

[85] , wherein the aluminum oxide is neutral aluminum oxide.

[0241]

[89] The method of paragraph

[84] , wherein the resin comprises silica.

[0242]

[90] The method of any one of paragraphs

[84] to

[89] , wherein the resin has a bulk density of 0.1 g / mL to 0.75 g / mL.

[0243]

[91] The method of paragraph

[90] , wherein the resin has a bulk density of 0.25 g / mL to 0.5 g / mL.

[0244]

[92] The method of any one of paragraphs

[84] to

[91] , wherein the resin has a pore volume of 0.1 mL / g to 1.5 mL / g.

[0245]

[93] The method of paragraph

[92] , wherein the resin has a pore volume of 0.5 mL / g to 1 mL / g.

[0246]

[94] The method of any one of paragraphs

[84] to

[93] , wherein the resin has a particle distribution size of 25 μm to 800 μm, and optionally, the resin has a particle distribution size of 100 μm to 800 μm.

[0247]

[95] The method of paragraph

[94] , wherein the resin has a particle distribution size of 300 μm to 600 μm.

[0248]

[96] The method of any one of paragraphs

[84] to

[95] , wherein the resin has an approximate moisture content of 2% (w / w) to 9% (w / w).

[0249]

[97] The method of paragraph

[96] , wherein the resin has an approximate moisture content of 4.5% (w / w) to 6.5% (w / w).

[0250]

[98] The method of any one of paragraphs

[84] to

[97] , wherein the preliminary liquid passing step includes passing a solvent through the resin to completely wet the resin.

[0251]

[99] The method according to paragraph

[98] , wherein the preliminary liquid passing step comprises passing the solvent through the resin in an amount of 1BV to 4BV.

[0252]

[0100] The method described in paragraph

[99] , wherein the preliminary flow step includes flowing the solvent through the resin in an amount of 2BV.

[0253]

[0101] A method according to any one of paragraphs

[98] to

[0100] , wherein the solvent is passed through the resin in the preliminary passing step with a minimum residence time in the column of at least 10 minutes.

[0254]

[0102] The method according to any one of paragraphs

[98] to

[0101] , wherein the solvent is passed through the resin at a maximum downward superficial velocity of 0.5 cm / min to 5 cm / min in the preliminary passing step.

[0255]

[0103] The method of paragraph

[0102] , wherein the solvent is passed through the resin in the preliminary passing step at a maximum downward superficial velocity of 1 cm / min to 4 cm / min.

[0256]

[0104] The method of any one of paragraphs

[98] to

[0103] , wherein the solvent is passed through the resin at a maximum upward superficial velocity of 0.05 cm / min to 5 cm / min.

[0257]

[0105] The method of paragraph

[0104] , wherein the solvent is passed through the resin at a maximum upflow superficial velocity of 0.1 cm / min to 3 cm / min.

[0258]

[0106] The method of any one of paragraphs

[84] to

[0105] , wherein the isoprenoid composition is exposed to the resin after the preliminary liquid passage step.

[0259]

[0107] The method of paragraph

[0106] , wherein the isoprenoid composition is exposed to the resin for a minimum residence time on the resin of at least 10 minutes.

[0260]

[0108] The method of paragraph

[0106] or

[0107] , wherein the isoprenoid composition is exposed to the resin at a maximum downward flow superficial velocity of 1 cm / min to 15 cm / min.

[0261]

[0109] The method of paragraph

[0108] , wherein the isoprenoid composition is exposed to the resin at a maximum downward flow superficial velocity of 5 cm / min to 11 cm / min.

[0262]

[0110] The method of any one of paragraphs

[0106] to

[0109] , wherein the isoprenoid composition is exposed to the resin at a maximum upward superficial velocity of 0.1 cm / min to 10 cm / min.

[0263]

[0111] The method of paragraph

[0110] , wherein the isoprenoid composition is exposed to the resin at a maximum upward superficial velocity of 1 cm / min to 5 cm / min.

[0264]

[0112] The method according to any one of paragraphs

[80] to

[0109] , wherein steps (a) to (c) are repeated 2 to 20 times.

[0265]

[0113] The method of paragraph

[0112] , wherein steps (a) to (c) are repeated 2 to 20 times.

[0266]

[0114] The method of paragraph

[0113] , wherein steps (a) to (c) are repeated 2 to 10 times.

[0267]

[0115] The method of paragraph

[0114] , wherein steps (a) to (c) are repeated six times.

[0268]

[0116] The method of any one of paragraphs

[84] to

[0115] , wherein the resin is regenerated after recovery of the isoprenoid from the resin in step (c).

[0269]

[0117] The resin (a) adding a polar solvent to the resin; (b) removing the polar solvent from the resin; (c) heating the resin; and (d) cooling the resin. The method described in paragraph

[0116] , which is reproduced by

[0270]

[0118] The method of paragraph

[0117] , wherein step (a) is carried out by adding a polar solvent to the resin in an amount of 1 BV to 10 BV.

[0271]

[0119] The method of paragraph

[0118] , wherein the polar solvent is added to the resin in an amount of 2BV to 6BV.

[0272]

[0120] The method of paragraph

[0119] , wherein the polar solvent is added to the resin in an amount of about 4 BV.

[0273]

[0121] The method of any one of paragraphs

[0117] to

[0120] , wherein step (c) is carried out by heating the resin to a temperature of from 100°C to 200°C.

[0274]

[0122] The method of paragraph

[0121] , wherein the resin is heated to a temperature of about 150° C.

[0275]

[0123] The method of any one of paragraphs

[0117] to

[0122] , wherein the cooling in step (d) is carried out by cooling the resin to room temperature.

[0276]

[0124] The method of any one of paragraphs

[73] to

[0123] , wherein the chromatography is carried out under N2.

[0277]

[0125] The isoprenoid is C5-C 60 The method according to any one of paragraphs

[73] to

[0124] , wherein the compound is an isoprenoid.

[0278]

[0126] The isoprenoid is C 15 -C 60 The method of paragraph

[0125] , wherein the compound is an isoprenoid.

[0279]

[0127] The method of any one of paragraphs

[73] to

[0126] , wherein the isoprenoid is a hemiterpenoid, monoterpenoid, sesquiterpenoid, diterpenoid, sesterterpenoid, triterpenoid, tetraterpenoid, or polyterpenoid.

[0280]

[0128] The method of claim

[0127] , wherein the isoprenoid is a sesquiterpenoid.

[0281]

[0129] The method described in paragraph

[0127] , wherein the isoprenoid is a monoterpenoid.

[0282]

[0130] The method of any one of paragraphs

[73] to

[0126] , wherein the isoprenoid is a hemiterpene, monoterpene, sesquiterpene, diterpene, sesterterpene, triterpene, tetraterpene, or polyterpene.

[0283]

[0131] The method of paragraph

[0130] , wherein the isoprenoid is a sesquiterpene.

[0284]

[0132] The method of any one of paragraphs

[73] to

[0124] , wherein the isoprenoid is abietadiene, amorphadiene, cadinane, carene, cuminaldehyde, eugenol, farnesene, geranial, isoprene, limonene, myrcene, ocimene, α-pinene, β-pinene, sabinene, γ-terpinene, terpinolene, thujone, neral, eucalyptol, citronellal, carvone, or valencene.

[0285]

[0133] The method of paragraph

[0132] , wherein the isoprenoid is farnesene.

[0286]

[0134] The method of paragraph

[0132] , wherein the isoprenoid is a cadinane.

[0287]

[0135] A method according to any one of paragraphs

[73] to

[0134] , wherein the isoprenoid is purified from the isoprenoid composition to a purity of about 95% (w / w) to about 100% (w / w).

[0288]

[0136] The method of paragraph

[0135] , wherein the isoprenoid is purified from the isoprenoid composition to a purity of about 98% (w / w) to about 100% (w / w).

[0289]

[0137] The method of paragraph

[0136] , wherein the isoprenoid is purified from the isoprenoid composition to a purity of about 99% (w / w) to about 100% (w / w).

[0290]

[0138] The method of paragraph

[0137] , wherein the isoprenoid composition is purified to a purity of about 99.5% (w / w) to about 100% (w / w).

[0291]

[0139] A composition comprising an isoprenoid, the composition being produced by a method described in any one of paragraphs

[73] to

[0138] .

[0292]

[0140] The composition described in paragraph

[0139] , wherein the isoprenoid has a purity of about 95% (w / w) to about 100% (w / w).

[0293]

[0141] The composition described in paragraph

[0139] , wherein the isoprenoid has a purity of about 98% (w / w) to about 100% (w / w), and optionally, the isoprenoid is purified from the isoprenoid composition to a purity of about 99% (w / w) to about 100% (w / w).

[0294]

[0142] The composition described in paragraph

[0141] , wherein the isoprenoid has a purity of about 99.5% (w / w) to about 100% (w / w).

[0295]

[0143] A composition described in any one of paragraphs

[0139] to

[0142] , containing one or more impurities including farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0296]

[0144] A composition comprising an isoprenoid and one or more impurities, wherein the purity of the isoprenoid is about 90% (w / w) to about 100% (w / w), and the one or more impurities include farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

[0297]

[0145] A composition described in paragraph

[0143] or

[0144] , comprising farnesol and having a farnesol concentration of 0 mg / mL to 1 g / L.

[0298]

[0146] The composition described in paragraph

[0145] , wherein the concentration of farnesol is 0 mg / L to 600 mg / L.

[0299]

[0147] The composition described in paragraph

[0146] , wherein the concentration of farnesol is less than 500 mg / L.

[0300]

[0148] A composition described in any one of paragraphs

[0143] to

[0147] , comprising farnesene epoxide and having a farnesol concentration of 0 mg / mL to 500 mg / L.

[0301]

[0149] The composition of paragraph

[0148] , wherein the concentration of the farnesene epoxide is 0 mg / L to 200 mg / L.

[0302]

[0150] The composition described in paragraph

[0149] , wherein the concentration of the farnesene epoxide is less than 150 mg / L.

[0303]

[0151] A composition comprising an isoprenoid and one or more carriers, diluents, or excipients, wherein the purity of the isoprenoid is from about 99.5% (w / w) to about 100% (w / w).

[0304]

[0152] The composition described in paragraph

[0151] , wherein the isoprenoid is present together with one or more impurities, and the one or more impurities are present at a concentration of about 0.5% (w / w) or less.

[0305]

[0153] The composition described in paragraph

[0152] , wherein the one or more impurities are present at a concentration of about 0.4% (w / w) or less, optionally, the one or more impurities are present at a concentration of about 0.3% (w / w) or less, optionally, the one or more impurities are present at a concentration of about 0.2% (w / w) or less, and optionally, the one or more impurities are present at a concentration of about 0.1% (w / w) or less.

[0306]

[0154] The composition of paragraph

[0152] or

[0153] , wherein the one or more impurities include farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

Claims

1. 1. A method for purifying farnesene, comprising: (a) providing a farnesene composition; and (b) purifying the farnesene from the farnesene composition of (a) by chromatography, wherein the chromatography comprises: (1) Preliminary passage of the resin; (2) exposing the farnesene composition to the resin; and (3) recovering the farnesene from the resin. The method comprising:

2. 10. The method of claim 1, wherein the farnesene composition comprises farnesene and one or more impurities.

3. 3. The method of claim 2, wherein the one or more impurities comprise one or more polar impurities including farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acids, sterols, hexahydrofarneseic acid (HHFA), santalene, or santalol.

4. 3. The method of claim 2, wherein the farnesene composition has a concentration of one or more impurities from 1 ppm to 15,000 ppm.

5. The resin is (a) a bulk density of 0.1 g / mL to 0.75 g / mL; (b) a pore volume of 0.1 mL / g to 1.5 mL / g; (c) a particle distribution size of 25 μm to 800 μm; or 10. The method of claim 1, wherein (d) the granules have an approximate moisture content of 0.01% w / w to 9% w / w.

6. 10. The method of claim 1, wherein the farnesene composition is exposed to the resin after the pre-bleeding step.

7. 7. The method of claim 6, wherein the farnesene composition is exposed to the resin at a minimum residence time on the resin of at least 10 minutes, a maximum downflow superficial velocity of from 1 cm / min to 15 cm / min, and a maximum upflow superficial velocity of from 0.1 cm / min to 10 cm / min.

8. 2. The method of claim 1, wherein steps (a) to (c) are repeated 2 to 20 times.

9. 10. The method of claim 1, wherein the resin is regenerated after recovery of the farnesene from the resin in step (c).

10. The resin is (a) adding a polar solvent to the resin; (b) removing the polar solvent from the resin; (c) heating the resin; and 10. The method of claim 9, wherein (d) the resin is regenerated by cooling.

11. 10. The method of claim 1, wherein the farnesene is purified from the farnesene composition to a purity of about 95% (w / w) to about 100% (w / w).

12. 1. A method for purifying an isoprenoid, comprising: (a) providing an isoprenoid composition; and (b) purifying the isoprenoid from the isoprenoid composition of (a) by chromatography, wherein the chromatography comprises: (1) Preliminary passage of the resin; (2) exposing the isoprenoid composition to the resin; and (3) recovering the isoprenoid from the resin.

13. 13. The method of claim 12, wherein the isoprenoid composition comprises farnesene and one or more impurities.

14. 14. The method of claim 13, wherein the one or more impurities comprise one or more polar impurities, optionally the one or more polar impurities comprise farneseic acid, farnesol, farnesene oxide, farnesene epoxide, fatty acid, sterol, HHFA, santalene, or santalol.

15. 14. The method of claim 13, wherein the isoprenoid composition has a concentration of one or more impurities from 1 ppm to 15,000 ppm.

16. The resin is (a) a bulk density of 0.1 g / mL to 0.75 g / mL; (b) a pore volume of 0.1 mL / g to 1.5 mL / g; (c) a particle distribution size of 25 μm to 800 μm; or 13. The method of claim 12, wherein (d) the mixture has an approximate moisture content of 0.01% w / w to 9% w / w.

17. 13. The method of claim 12, wherein the isoprenoid composition is exposed to the resin at a minimum residence time on the resin of at least 10 minutes, a maximum downflow superficial velocity of 1 cm / min to 15 cm / min, and a maximum upflow superficial velocity of 0.1 cm / min to 10 cm / min.

18. 13. The method of claim 12, wherein steps (a) to (c) are repeated 2 to 20 times.

19. 13. The method of claim 12, wherein the resin is regenerated after recovery of the isoprenoid from the resin in step (c).

20. The resin is (e) adding a polar solvent to the resin; (f) removing the polar solvent from the resin; (g) heating the resin; and 20. The method of claim 19, wherein (h) the resin is regenerated by cooling.

21. 13. The method of claim 12, wherein the isoprenoid is abietadiene, amorphadiene, cadinane, carene, cuminaldehyde, eugenol, farnesene, geranial, isoprene, limonene, myrcene, ocimene, α-pinene, β-pinene, sabinene, γ-terpinene, terpinolene, thujone, neral, eucalyptol, citronellal, carvone, or valencene.

22. 13. The method of claim 12, wherein the isoprenoid is a cadinane.

23. 13. The method of claim 12, wherein the isoprenoid is farnesene.

24. 13. The method of claim 12, wherein the isoprenoid is purified from the isoprenoid composition to a purity of about 95% (w / w) to about 100% (w / w).

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