The present invention concerns the field of recombinant manufacture of santalene and related products. In particular, it relates to a method for the manufacture of a composition comprising at least one santalene comprising the step of converting farnesyl pyrophosphate into at least one santalene, wherein said conversion is carried out by a polypeptide exhibiting santalene synthase activity. Moreover, the invention contemplates a composition comprising a mixture of beta-santalene and alpha-santalene with an excess of beta-santalene obtainable by the method of the invention. The invention also relates to the use of a santalene synthase polypeptide, a heterologous polynucleotide encoding it, a vector or gene construct comprising said polynucleotide, a host cell or a non-human transgenic organism comprising said gene construct or vector for the manufacture of a composition comprising at least one santalene, preferably, beta-santalene, more preferably a mixture of beta-santalene and alpha-santalene. Further, the present invention relates to a method for manufacturing a composition comprising at least one santalol, preferably beta-santalol, comprising producing a composition comprising at least one santalene by the aforementioned method of the invention and oxidising said at least one santalene, preferably, beta-santalene, into the respective alcohol in order to manufacture a composition comprising at least one santalol, preferably, beta-santalol. Yet, the invention pertains to a kit for the manufacture of a composition comprising at least one santalene comprising the aforementioned polypeptide. heterologous polynucleotide, vector or gene construct, host cell or non-human transgenic organism and to a non-human host cell or non-human transgenic organism expressing a polypeptide exhibiting santalene synthase activity from the aforementioned heterologous polynucleotide. vector or gene construct.
An artificial alkane oxidation system comprising components: a. an oxidase enzyme; and b. one or more enzymes to provide one or more alkanes. The artificial alkane oxidation system optionally has c. an electron transfer compound suitable to transfer at least one electron to the oxidase enzyme; and further optionally has d. an electron transfer compound regeneration enzyme suitable to reduce the electron transfer compound of c. when it is in its oxidized state. The oxidase enzyme is an amino acid sequence with a sequence identity of at least 70% with any one of SEQ ID NO: 1, 10, 11, 23 to 43 or a fragment thereof or a variant thereof.
The present invention refers to a method for producing a zizaene-containing composition comprising the following subsequent steps: i) providing farnesyl pyrophosphate and at least one polypeptide exhibiting zizaene synthase activity under conditions suitable for the polypeptide to convert at least a portion of the farnesyl pyrophosphate into zizaene; ii) converting, by means of the polypeptide, at least a portion of the farnesyl pyrophosphate into a bisabolyl cation; and iii) producing a zizaene-containing composition by converting, by means of the polypeptide, the bisabolyl cation into zizaene and, optionally, at least one further compound, wherein the at least one polypeptide comprises an amino acid sequence selected from the group consisting of: a) an amino acid sequence as shown in SEQ ID NO: 1; b) an amino acid sequence which is at least 55 % identical to the amino acid sequences as shown in SEQ ID NO: 1; c) an amino acid sequence encoded by a nucleic acid sequence as shown in any one of SEQ ID NOs: 2, 3 or 5; d) an amino acid sequence encoded by a nucleic acid sequence which is at least 55 % identical to a nucleic acid sequence as shown in any one of SEQ ID NOs: 2, 3 or 5; and e) an amino acid sequence being a fragment of any one of the sequences listed in a) to d). Moreover, the invention is, inter alia, directed to a zizaene- containing composition obtainable with said method as well as to an according non-human host cell and an according polypeptide exhibiting zizaene synthase activity.
The present invention relates to an alcohol acyl transferase which is capable of esterifying a tertiary monoterpene alcohol such that at least 30% by mass of said tertiary monoterpene alcohol is esterified, preferably within 36 h, 24 h, 18 h, 12 h, 6 h, 3 h, 2 h, 1 h, 45 min or 30 min, more preferably in a microbial cell. The invention further relates to a nucleic acid comprising a nucleic acid sequence encoding the alcohol acyl transferase of the invention, or a complementary sequence thereof, and a vector or gene construct comprising the nucleic acid of the invention. Further provided by the present invention is a host cell comprising the vector or gene construct of the invention, and a transgenic non-human organism comprising the nucleic acid of the invention, the vector or gene construct of the invention, or the host cell of the invention. The invention also concerns a method for preparing a monoterpene ester, comprising esterifying a monoterpene alcohol to a monoterpene ester, in the presence of an alcohol acyl transferase of the invention. Specifically, it provides a method for preparing linalyl acetate, comprising esterifying linalool to linalyl acetate, in the presence of an alcohol acyl transferase of the invention. The invention further pertains to the use of the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene construct of the invention, the host cell of the invention, or the transgenic non-human organism of the invention (i) for heterologous reconstitution of a terpene biosynthetic pathway; (ii) for producing an industrial product, preferably a flavour or fragrance, a biofuel, a fuel composition, a fuel compound, e.g., a blowing agent for diesel fuel compositions, a pesticide, an insect repellent or an antimicrobial; (ill) for producing an aliphatic and/or aromatic monoterpene ester from a monoterpene alcohol, preferably from a tertiary monoterpene alcohol; (iv) for detoxifying a monoterpene alcohol in a microorganism, thereby increasing monoterpene production in said microorganism; (v) in combination with a GPP synthase and/or S- or R-linalool synthase; (vi) for increasing the beneficial effects of acetylation in that the hydrophobic acetate partitions more readily go into an organic phase, as compared to the monoterpene alcohol; (vii) for expressing the alcohol acyl transferase of the invention such that the ratio of monoterpene acetate to monoterpene alcohol is greater than 5:1 or 10:1 or (viii) in a microbial production system for monoterpene esters. The invention also provides a kit comprising the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene construct of the invention, the host cell of the invention, or the transgenic non-human organism of the invention, and optionally at least one monoterpene alcohol, preferably a tertiary monoterpene alcohol. Finally, the invention relates to a method for the production of a fuel and/or biolubricant compound, wherein the method comprises the steps of: a) Producing one or more monoterpene esters by any one of the methods of the invention; b) optionally, purifying the one or more monoterpene esters produced in step a); and c) converting part or all of the one or more monoterpene esters of step a), or the optionally purified one or more monoterpene esters of step b), to one or more fuel and/or biolubricant compound, preferably selected from the group consisting of: tetrahydrolinalool; 2,6-dimethyloctane (DMO); saturated C20 hydrocarbon dimers; saturated C30 hydrocarbon trimers; hydrogenated methylcyclopentadiene dimers; saturated high density multi-cyclic hydrocarbon compounds suitable for missile propulsion; and hydrogenated C40+ oligomers suitable to produce biolubricant additives; d) optionally combining the one or more fuel or biolubricant compound with additional compounds suitable for a fuel and/or biolubricant; wherein the fuel and/or biolubricant composition has in sum between and including 0.01% (w/w) to 99.99% (w/w), of the fuel or biolubricant compound produced from the one or more monoterpene esters obtainable by one of the methods of the invention.
The present invention relates to a sensing composition for detecting a volatile organic compound in its gas phase, comprising 1) a silylated cellulose selected from the group of trialkylsilyl cellulose, triarylsilyl cellulose and alkylarylsilyl cellulose; and 2) a fluorescent dye. The detection of a volatile with this sensing composition comprises placing the sensing composition (optionally present as a layer on a support) in the vicinity of a potential source of a volatile and analyzing the sensing composition to determine whether a volatile of the potential source has reached the sensing composition. This detection can be performed in a quantitative manner by means of spectrophotometric analysis, and/or by image analysis of the sensing composition. The invention is particularly useful for application in high throughput screening.
Disclosed are santalene synthases with improved product profile and methods for improving santalene synthases. The invention further relates to santalene compositions produced by fermentation that have a greater beta-santalene content than alpha-santalene content.
The present invention concerns the field of recombinant manufacture of santalene and related products. In particular, it relates to a method for the manufacture of a composition comprising at least one santalene comprising the step of converting farnesyl pyrophosphate into at least one santalene, wherein said conversion is carried out by a polypeptide exhibiting santalene synthase activity. Moreover, the invention contemplates a composition comprising a mixture of beta-santalene and alpha-santalene with an excess of beta-santalene obtainable by the method of the invention. The invention also relates to the use of a santalene synthase polypeptide, a heterologous polynucleotide encoding it, a vector or gene construct comprising said polynucleotide, a host cell or a non-human transgenic organism comprising said gene construct or vector for the manufacture of a composition comprising at least one santalene, preferably, beta-santalene, more preferably a mixture of beta-santalene and alpha-santalene. Further, the present invention relates to a method for manufacturing a composition comprising at least one santalol, preferably beta-santalol, comprising producing a composition comprising at least one santalene by the aforementioned method of the invention and oxidising said at least one santalene, preferably, beta-santalene, into the respective alcohol in order to manufacture a composition comprising at least one santalol, preferably, beta-santalol. Yet, the invention pertains to a kit for the manufacture of a composition comprising at least one santalene comprising the aforementioned polypeptide, heterologous polynucleotide, vector or gene construct, host cell or non-human transgenic organism and to a non-human host cell or non- human transgenic organism expressing a polypeptide exhibiting santalene synthase activity from the aforementioned heterologous polynucleotide, vector or gene construct.
An artificial alkane oxidation system comprising components: a. an oxidase enzyme; and b. one or more enzymes to provide one or more alkanes. The artificial alkane oxidation system optionally has c. an electron transfer compound suitable to transfer at least one electron to the oxidase enzyme; and further optionally has d. an electron transfer compound regeneration enzyme suitable to reduce the electron transfer compound of c. when it is in its oxidized state. The oxidase enzyme is an amino acid sequence with a sequence identity of at least 70 % with any one of SEQ ID NO: 1, 10, 11, 23 to 43 or a fragment thereof or a variant thereof.
Disclosed is a method for the manufacture of at least one C-20 terpenoid alcohol comprising the steps of converting geranylgeranyl pyrophosphate into copalyl diphosphate (CPP) or labda-13-en-8-ol diphosphate (LPP) and converting CPP or LPP into at least one C-20 terpenoid alcohol, wherein said conversion is carried out by a polypeptide exhibiting diterpene alcohol synthase activity capable of converting CPP into manool, LPP into sclareol and/or LPP into abienol, and wherein said polypeptide comprises an amino acid sequence as specified in the claims. The invention further relates to the aforementioned polypeptide exhibiting diterpene alcohol synthase activity as well as a fusion protein comprising said polypeptide, a polynucleotide encoding it, a vector or gene construct comprising said polynucleotide, a host cell comprising said vector or gene construct, a non-human transgenic organism comprising the polynucleotide, vector, gene construct or host cell, as well as uses thereof for the manufacture of at least one C-20 terpenoid alcohol.
C12Q 1/527 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving lyase
The invention relates to a process for the oxidation of santalene to santalol. The starting material is in particular a mixture comprising alpha-santalene, beta-santalene, epi-beta-santalene, trans-alpha-bergamotene and beta-bisabolene. The oxidation of the santalenes occurs via an intermediate chloro-santalene compound. Substitution of the chloro-substituent by acetate yielded the mixture of the corresponding santalyl acetates, which were hydrolyzed to yield the corresponding mixture of santalols.
C07C 29/09 - Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring by hydrolysis
C07C 17/10 - Preparation of halogenated hydrocarbons by replacement by halogens of hydrogen atoms
C07C 22/02 - Cyclic compounds containing halogen atoms bound to an acyclic carbon atom having unsaturation in the rings
C07C 67/11 - Preparation of carboxylic acid esters by reacting carboxylic acids or symmetrical anhydrides with ester groups or with a carbon-halogen bond being mineral ester groups
The invention is directed to a santalene synthase, to a nucleic acid encoding said santalene synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing santalene, to a method of preparing santalol and to a method of preparing a santalene synthase. The invention is further directed to an antibody specific for the santalane synthase.
The present invention relates to an alcohol acyl transferase which is capable of esterifying a tertiary monoterpene alcohol such that at least 30% by mass of said tertiary monoterpene alcohol is esterified, preferably within 36 h, 24 h, 18 h, 12 h, 6 h, 3 h, 2 h, 1 h, 45 min or 30 min, more preferably in a microbial cell. The invention further relates to a nucleic acid comprising a nucleic acid sequence encoding the alcohol acyl transferase of the invention, or a complementary sequence thereof, and a vector or gene construct comprising the nucleic acid of the invention. Further provided by the present invention is a host cell comprising the vector or gene construct of the invention, and a transgenic non-human organism comprising the nucleic acid of the invention, the vector or gene construct of the invention, or the host cell of the invention. The invention also concerns a method for preparing a monoterpene ester, comprising esterifying a monoterpene alcohol to a monoterpene ester, in the presence of an alcohol acyl transferase of the invention. Specifically, it provides a method for preparing linalyl acetate, comprising esterifying linalool to linalyl acetate, in the presence of an alcohol acyl transferase of the invention. The invention further pertains to the use of the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene construct of the invention, the host cell of the invention, or the transgenic non-human organism of the invention (i) for heterologous reconstitution of a terpene biosynthetic pathway; (ii) for producing an industrial product, preferably a flavour or fragrance, a biofuel, a fuel composition, a fuel compound, e.g., a blowing agent for diesel fuel compositions, a pesticide, an insect repellent or an antimicrobial; (ill) for producing an aliphatic and/or aromatic monoterpene ester from a monoterpene alcohol, preferably from a tertiary monoterpene alcohol; (iv) for detoxifying a monoterpene alcohol in a microorganism, thereby increasing monoterpene production in said microorganism; (v) in combination with a GPP synthase and/or S- or R- linalool synthase; (vi) for increasing the beneficial effects of acetylation in that the hydrophobic acetate partitions more readily go into an organic phase, as compared to the monoterpene alcohol; (vii) for expressing the alcohol acyl transferase of the invention such that the ratio of monoterpene acetate to monoterpene alcohol is greater than 5:1 or 10:1 or (viii) in a microbial production system for monoterpene esters. The invention also provides a kit comprising the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene construct of the invention, the host cell of the invention, or the transgenic non-human organism of the invention, and optionally at least one monoterpene alcohol, preferably a tertiary monoterpene alcohol. Finally, the invention relates to a method for the production of a fuel and/or biolubricant compound, wherein the method comprises the steps of: a) Producing one or more monoterpene esters by any one of the methods of the invention; b) optionally, purifying the one or more monoterpene esters produced in step a); and c) converting part or all of the one or more monoterpene esters of step a), or the optionally purified one or more monoterpene esters of step b), to one or more fuel and / or biolubricant compound, preferably selected from the group consisting of: tetrahydrolinalool; 2,6-dimethyloctane (DMO); saturated C20 hydrocarbon dimers; saturated C30 hydrocarbon trimers; hydrogenated methylcyclopentadiene dimers; saturated high density multi-cyclic hydrocarbon compounds suitable for missile propulsion; and hydrogenated C40+ oligomers suitable to produce biolubricant additives; d) optionally combining the one or more fuel or biolubricant compound with additional compounds suitable for a fuel and / or biolubricant; wherein the fuel and / or biolubricant composition has in sum between and including 0.01 % (w/w) to 99.99 % (w/w), of the fuel or biolubricant compound produced from the one or more monoterpene esters obtainable by one of the methods of the invention.
Disclosed are santalene synthases with improved product profile and methods for improving santalene synthases. The invention further relates to santalene compositions produced by fermentation that have a greater beta-santalene content than alpha-santalene content.
The invention is directed to a patchoulol synthase, to a nucleic acid encoding said patchoulol synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing patchoulol and elemol, and preferably also pogostol, and to a method of preparing a patchoulol synthase.
The invention relates to a process for the oxidation of santalene to santalol. The starting mate- rial is in particular a mixture comprising alpha-santalene, beta-santalene, epi-beta-santalene, trans-alpha-bergamotene and beta-bisabolene. The oxidation of the santalenes occurs via an intermediate chloro-santalene compound. Substitution of the chloro-substituent by acetate yielded the mixture of the corresponding santalyl actates, which were hydrolyzed to yield the corresponding mixture of santalols.
C07C 33/14 - Alcohols containing rings other than six-membered aromatic rings containing six-membered rings
C07C 29/09 - Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring by hydrolysis
C07C 17/10 - Preparation of halogenated hydrocarbons by replacement by halogens of hydrogen atoms
C07C 22/00 - Cyclic compounds containing halogen atoms bound to an acyclic carbon atom
C07C 22/02 - Cyclic compounds containing halogen atoms bound to an acyclic carbon atom having unsaturation in the rings
C07C 67/11 - Preparation of carboxylic acid esters by reacting carboxylic acids or symmetrical anhydrides with ester groups or with a carbon-halogen bond being mineral ester groups
C07C 69/145 - Acetic acid esters of monohydroxylic compounds of unsaturated alcohols
The present invention relates to a sensing composition for detecting a volatile organic compound in its gas phase, comprising 1) a silylated cellulose selected from the group of trialkylsilyl cellulose, triarylsilyl cellulose and alkylarylsilyl cellulose; and 2) a fluorescent dye. The detection of a volatile with this sensing composition comprises placing the sensing composition (optionally present as a layer on a support) in the vicinity of a potential source of a volatile and analyzing the sensing composition to determine whether a volatile of the potential source has reached the sensing composition. This detection can be performed in a quantitative manner by means of spectrophotometric analysis, and/or by image analysis of the sensing composition. The invention is particularly useful for application in high throughput screening.
G01N 31/22 - Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroupsApparatus specially adapted for such methods using chemical indicators
G01N 21/78 - Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
01 - Chemical and biological materials for industrial, scientific and agricultural use
03 - Cosmetics and toiletries; cleaning, bleaching, polishing and abrasive preparations
30 - Basic staples, tea, coffee, baked goods and confectionery
Goods & Services
Chemicals used in industry, science and photography, as well
as in agriculture, horticulture and forestry; unprocessed
artificial resins, unprocessed plastics; manures; fire
extinguishing compositions; tempering and soldering
preparations; chemical substances for preserving foodstuffs;
tanning substances; adhesives used in industry; chemical
preparations for foods and the perfume industry. Natural aromas and fragrances for the perfume industry. Natural aromas and fragrances for food.
01 - Chemical and biological materials for industrial, scientific and agricultural use
03 - Cosmetics and toiletries; cleaning, bleaching, polishing and abrasive preparations
30 - Basic staples, tea, coffee, baked goods and confectionery
Goods & Services
Chemicals used in industry; chemical preparations for foods and the perfume industry Natural aromas in the nature of oils for perfumes and scents and fragrances for the perfume industry Natural aromas and fragrances for food, namely, flavorings and extractions in the nature of extracts used as food flavoring, other than essential oils
01 - Chemical and biological materials for industrial, scientific and agricultural use
03 - Cosmetics and toiletries; cleaning, bleaching, polishing and abrasive preparations
30 - Basic staples, tea, coffee, baked goods and confectionery
Goods & Services
Chemicals used in industry, science and photography, as well as in agriculture, horticulture and forestry; unprocessed artificial resins, unprocessed plastics; manures; fire extinguishing compositions; tempering and soldering preparations; chemical substances for preserving foodstuffs; tanning substances; adhesives used in industry; chemical preparations for foods and the perfume industry. Natural aromas and fragrances for the perfume industry. Natural aromas and fragrances for food.
01 - Chemical and biological materials for industrial, scientific and agricultural use
03 - Cosmetics and toiletries; cleaning, bleaching, polishing and abrasive preparations
30 - Basic staples, tea, coffee, baked goods and confectionery
Goods & Services
Chemicals used in industry, science and photography, as well as in agriculture, horticulture and forestry, except fungicides, herbicides, insecticides and parasiticides; unprocessed artificial resins, unprocessed plastics; manures; fire extinguishing compositions; tempering and soldering preparations; chemical substances for preserving foodstuffs; adhesives used in industry; chemical preparations for foods and the perfume industry Natural aromas in the nature of oils for perfumes and scents and fragrances for the perfume industry Natural aromas and fragrances for food, namely, flavorings and extractions in the nature of extracts used as food flavoring, other than essential oils
The invention is directed to a santalene synthase, to a nucleic acid encoding said santalene synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing santalene, to a method of preparing santalol and to a method of preparing a santalene synthase. The invention is further directed to an antibody specific for the santalane synthase.
The invention relates to a two phase fermentation process for producing an organic compound, in particular an isoprenoid and to a bioreactor comprising a two phase fermentation system for producing an organic compound.
The invention is directed to a patchoulol synthase, to a nucleic acid encoding said patchoulol synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing patchoulol and elemol, and preferably also pogostol, and to a method of preparing a patchoulol synthase.
The invention is directed to a santalene synthase, to a nucleic acid encoding said santalene synthase, to an expression vector comprising said nucleic acid, to a host cell comprising said expression vector, to a method of preparing santalene, to a method of preparing santalol and to a method of preparing a santalene synthase. The invention is further directed to an antibody specific for the santalane synthase.
The invention relates to a two phase fermentation process for producing an organic compound, in particular an isoprenoid and to a bioreactor comprising a two phase fermentation system for producing an organic compound.
The invention relates to a Rhodobacter host cell, comprising a nucleic acid encoding - enzymes of a mevalonate pathway for making isoprenyl pyrophosphate (IPP) and its isomer dimethylallyl pyrophosphate (DMAPP); - an enzyme having catalytic activity for the condensation of IPP and DMAPP into geranyl diphosphate (GPP) and - an enzyme having monoterpene synthase activity in the conversion of GPP into a monoterpene or sesquiterpene synthase activity.
The present invention relates to a novel valencene synthase, to a nucleic acid encoding such valencene synthase, to a host cell comprising said encoding nucleic acid sequence and to a method for preparing valencene, comprising converting farnesyl diphosphate to valencene in the presence of a valencene synthase according to the invention.
The present invention relates to a valencene synthase, to a nucleic acid encoding such valencene synthase, to a host cell comprising said encoding nucleic acid sequence and to a method for preparing valencene, comprising converting farnesyl diphosphate to valencene in the presence of a valencene synthase according to the invention.
01 - Chemical and biological materials for industrial, scientific and agricultural use
03 - Cosmetics and toiletries; cleaning, bleaching, polishing and abrasive preparations
30 - Basic staples, tea, coffee, baked goods and confectionery
Goods & Services
Chemicals used in industry, science and photography, as well as in agriculture, horticulture and forestry; unprocessed artificial resins, unprocessed plastics; manures; fire extinguishing compositions; tempering and soldering preparations; chemical substances for preserving foodstuffs; tanning substances; adhesives used in industry; chemical preparations for foods and the perfume industry. Natural aromas and fragrances for the perfume industry. Natural aromas and fragrances for food.
30.
PREPARATION OF 4-HYDROXY-2,5-DIMETHYL-2,3-DIHYDROFURAN-3-ONE
The present invention relates to a method for preparing a 4-hydroxy- 2,5-dimethyl-2,3-dihydrofuran-3-one or an analogue thereof, comprising reacting an α-hydroxyaldehyde or an α-ketoaldehyde (compound A) and an α-hydroxyketone (compound B) in the presence of an org a no-catalyst (C) capable of catalysing an aldol reaction, preferably selected from the group consisting of aldolase, proline, pyrrolidine and catalysts comprising a proline active site or a pyrrolidine active site, and forming 4-hydroxy-2,5-dimethyl-2,3-dihydrofuran-3-one or an analogue thereof.
The invention relates to a process for the preparation of &dgr;-lactones by reacting acetaldehyde and a substituted acetaldehyde in the presence of an aldolase, preferably a DERA from Escherichia coli, from Bacillus subtilis or from a Geobacillus species, &dgr;-lactones, such as saturated &dgr;-lactones, &agr;, &bgr;-unsaturated &dgr;-lactones and 3-hydroxy- &dgr;-lactones can suitably be used as (intermediates in the production of) flavors and/or fragrances and/or pharmaceuticals.
