- P450Jα: A New, Robust and α-Selective Fatty Acid Hydroxylase Displaying Unexpected 1-Alkene Formation
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Oxyfunctionalization of fatty acids (FAs) is a key step in the design of novel synthetic pathways for biobased/biodegradable polymers, surfactants and fuels. Here, we show the isolation and characterization of a robust FA α-hydroxylase (P450Jα) which catalyses the selective conversion of a broad range of FAs (C6:0-C16:0) and oleic acid (C18:1) with H2O2 as oxidant. Under optimized reaction conditions P450Jα yields α-hydroxy acids all with >95 % regioselectivity, high specific activity (up to 15.2 U mg?1) and efficient coupling of oxidant to product (up to 85 %). Lauric acid (C12:0) turned out to be an excellent substrate with respect to productivity (TON=394 min?1). On preparative scale, conversion of C12:0 reached 83 % (0.9 g L?1) when supplementing H2O2 in fed-batch mode. Under similar conditions P450Jα allowed further the first biocatalytic α-hydroxylation of oleic acid (88 % conversion on 100 mL scale) at high selectivity and in good yields (1.1 g L?1; 79 % isolated yield). Unexpectedly, P450Jα displayed also 1-alkene formation from shorter chain FAs (≤C10:0) showing that oxidative decarboxylation is more widely distributed across this enzyme family than reported previously.
- Armbruster, Julia,Steinmassl, Mathilde,Müller Bogotá, Christina A.,Berg, Gabriele,Nidetzky, Bernd,Dennig, Alexander
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p. 15910 - 15921
(2020/10/29)
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- Preparative Asymmetric Synthesis of Canonical and Non-canonical α-amino Acids Through Formal Enantioselective Biocatalytic Amination of Carboxylic Acids
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Chemical and biocatalytic synthesis of non-canonical α-amino acids (ncAAs) from renewable feedstocks and using mild reaction conditions has not efficiently been solved. Here, we show the development of a three-step, scalable and modular one-pot biocascade for linear conversion of renewable fatty acids (FAs) into enantiopure l-α-amino acids. In module 1, selective α-hydroxylation of FAs is catalyzed by the P450 peroxygenase P450CLA. By using an automated H2O2 supplementation system, efficient conversion (46 to >99%; TTN>3300) of a broad range of FAs (C6:0 to C16:0) into valuable α-hydroxy acids (α-HAs; >90% α-selective) is shown on preparative scale (up to 2.3 g L?1 isolated product). In module 2, a redox-neutral hydrogen borrowing cascade (alcohol dehydrogenase/amino acid dehydrogenase) allowed further conversion of α-HAs into l-α-AAs (20 to 99%). Enantiopure l-α-AAs (e.e. >99%) including the pharma synthon l-homo-phenylalanine can be obtained at product titers of up to 2.5 g L?1. Based on renewables and excellent atom economy, this biocascade is among the shortest and greenest synthetic routes to structurally diverse and industrially relevant ncAAs. (Figure presented.).
- Dennig, Alexander,Blaschke, Fabio,Gandomkar, Somayyeh,Tassano, Erika,Nidetzky, Bernd
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supporting information
(2019/02/09)
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- Preparative Asymmetric Synthesis of Canonical and Non-canonical a-amino Acids through Formal Enantioselective Biocatalytic Amination of Carboxylic Acids
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Chemical and biocatalytic synthesis of non-canonical a-amino acids (ncAAs) from renewable feedstocks and using mild reaction conditions has not efficiently been solved. Here, we show the development of a three-step, scalable and modular one-pot biocascade for linear conversion of renewable fatty acids (FAs) into enantiopure l-a-amino acids. In module 1, selective a-hydroxylation of FAs is catalyzed by the P450 peroxygenase P450CLA. By using an automated H2O2 supplementation system, efficient conversion (46 to >99%; TTN>3300) of a broad range of FAs (C6:0 to C16:0) into valuable a-hydroxy acids (a-HAs; >90% a-selective) is shown on preparative scale (up to 2.3 gL1 isolated product). In module 2, a redox-neutral hydrogen borrowing cascade (alcohol dehydrogenase/amino acid dehydrogenase) allowed further conversion of a-HAs into l-a-AAs (20 to 99%). Enantiopure l-a-AAs (e.e. >99%) including the pharma synthon l-homo-phenylalanine can be obtained at product titers of up to 2.5 gL1. Based on renewables and excellent atom economy, this biocascade is among the shortest and greenest synthetic routes to structurally diverse and industrially relevant ncAAs.
- Dennig, Alexander,Blaschke, Fabio,Gandomkar, Somayyeh,Tassano, Erika,Nidetzky, Bernd
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supporting information
p. 1348 - 1358
(2019/10/28)
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- α-Oxidative decarboxylation of fatty acids catalysed by cytochrome P450 peroxygenases yielding shorter-alkyl-chain fatty acids
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Cytochrome P450 peroxygenases belonging to the CYP152 family catalyse the oxidation of fatty acids using H2O2. CYP152N1 isolated from Exiguobacterium sp. AT1b exclusively catalyses the α-selective hydroxylation of myristic acid at physiological H2O2 concentration. However, a series of shorter-alkyl-chain fatty acids such as tridecanoic acid were produced from myristic acid by increasing the concentration of H2O2 (1-10 mM). The yield of tridecanoic acid from myristic acid reached 17%. An 18O-labeled oxidant study suggested that CYP152N1 catalysed the overoxidation of α-hydroxymyristic acid to form α-ketomyristic acid, which in turn was spontaneously decomposed by H2O2 to yield tridecanoic acid. Crystal structure analysis of CYP152N1 revealed its high similarity to other CYP152 family enzymes, such as CYP152A1 and CYP152B1. MD simulations of α-hydroxymyristic acid accommodated in CYP152N1 proposed a possible pre-oxidation conformation of α-hydroxymyristic acid for the decarboxylation reaction.
- Onoda, Hiroki,Shoji, Osami,Suzuki, Kazuto,Sugimoto, Hiroshi,Shiro, Yoshitsugu,Watanabe, Yoshihito
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p. 434 - 442
(2018/02/07)
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- In situ formation of H2O2 for P450 peroxygenases
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An in situ H2O2 generation approach to promote P450 peroxygenases catalysis was developed through the use of the nicotinamide cofactor analogue 1-benzyl-1,4-dihydronicotinamide (BNAH) and flavin mononucleotide (FMN). Final productivity could be enhanced due to higher enzyme stability at low H2O2 concentrations. The H2O2 generation represented the rate-limiting step, however it could be easily controlled by varying both FMN and BNAH concentrations. Further characterization can result in an optimized ratio of FMN/BNAH/O2/biocatalyst enabling high reaction rates while minimizing H2O2-related inactivation of the enzyme.
- Paul, Caroline E.,Churakova, Ekaterina,Maurits, Elmer,Girhard, Marco,Urlacher, Vlada B.,Hollmann, Frank
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p. 5692 - 5696
(2015/01/09)
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- Crystal structure of H2O2-dependent cytochrome P450SPαwith its bound fatty acid substrate: Insight into the regioselective hydroxylation of fatty acids at the α position
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Cytochrome P450SPα (CYP152B1) isolated from Sphingomonas paucimobilis is the first P450 to be classified as a 2O 2-dependent P450. P450SPα hydroxylates fatty acids with high α-regioselectivity. Herein we report the crystal structure of P450SPαwith palmitic acid as a substrate at a resolution of 1.65 A. The structure revealed that the Cα of the bound palmitic acid in one of the alternative conformations is 4.5 A from the heme iron. This conformation explains the highly selective α-hydroxylation of fatty acid observed in P450SPα. Mutations at the active site and the F-G loop of P450SPα did not impair its regioselectivity. The crystal structures of mutants (L78F and F288G) revealed that the location of the bound palmitic acid was essentially the same as that in the WT, although amino acids at the active site were replaced with the corresponding amino acids of cytochrome P450BSβ (CYP152A1), which shows β-regioselectivity. This implies that the high regioselectivity of P450SPα is caused by the orientation of the hydrophobic channel, which is more perpendicular to the heme plane than that of P450 BSβ.
- Fujishiro, Takashi,Shoji, Osami,Nagano, Shingo,Sugimoto, Hiroshi,Shiro, Yoshitsugu,Watanabe, Yoshihito
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experimental part
p. 29941 - 29950
(2012/04/10)
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- Nanohybrids composed of quantum dots and cytochrome P450 as photocatalysts
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(Chemical Equation Presented) Synthesis with light: CdS quantum dots (QDs) generate superoxide and hydroxyl radicals upon UV irradiation in aqueous solution. The radicals are used for activating P450BSβ enzymes attached at the QD surface, effecting the catalytic transformation of myristic acid into α- and β-hydroxymyristic acid (see picture, R = (CH 2)10CH3).
- Ipe, Binil Itty,Niemeyer, Christof M.
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p. 504 - 507
(2007/10/03)
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- Personal care compositions comprising solid particles enterapped in a gel network
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The present invention relates to a personal care composition comprising a three dimensional gel polymeric network comprising: a. a polymer; b. one or more solid particles that are entrapped within said polymer during polymerization of said polymer; and c. a solvent in which said polymer is dispersed. Another embodiment further includes at least one second colorant that is substantially similar to an at least one first colorant which is a solid particle and wherein said second colorant is dispersed within said composition but is not entrapped in said polymer and is separate and distinct from said network. In contrast, a third embodiment allows for the at least one second colorant to be substantially different from the at least one first colorant.
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- COSMETIC COMPOSITIONS CONTAINING A SILOXANE ELASTOMER
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A skin treatment composition is provided which includes a crosslinked non-emulsifying siloxane elastomer, a volatile siloxane and at least 50% by weight of water. Inclusion of the elastomer provides a unique liquid/powdery feel when rubbed into the skin.
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- Cosmetic compositions
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The present invention relates to pigmented emulsion cosmetic compositions containing emulsifying silicone elastomers that provide a natural appearance to the skin upon application. In particular, these cosmetic compositions are formulated such that agglomeration of the pigment upon application to the skin is minimized.
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- Cosmetic compositions
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A skin treatment composition is provided which includes a crosslinked siloxane elastomer gel of specific yield point, a skin-conditioning agent, a volatile siloxane and water. Inclusions of the select elastomers provide improved uniform distribution of the pigments.
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- Anhydrous cosmetic compositions
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An anhydrous skin treatment composition is provided which includes a crosslinked emulsifying siloxane elastomer, at least 20% humectant and a volatile siloxane. Inclusion of the elastomer provides a non-traditional smooth/silky feel to the skin upon application with a non-draggy rub in.
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- Cosmetic compositions
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The invention relates to cosmetic compositions comprising a combination of non-emulsifying and emulsifying crosslinked siloxane elastomers.
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- Anhydrous cosmetic compositions
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An anhydrous skin treatment composition is provided which includes a crosslinked siloxane elastomer gel of specific yield point, a skin conditioning agent and a volatile siloxane. Inclusions of the select elastomers provide improved uniform distribution of the pigments.
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- Cosmetic compositions
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The invention relates to cosmetic compositions comprising a combination of non-emulsifying and emulsifying crosslinked siloxane elastomers.
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- Role of the hydrophobic moiety of tumor promoters. Synthesis and activity of 2-alkylated benzolactams
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The size and position of a hydrophobic moiety on a benzolactam skeleton, which reproduces the active conformation and biological activity of teleocidins, play an important role in the appearance of the activity. Compounds with alkyl groups of various sizes and shapes at the 2-position of benzolactam were synthesized. Structure-activity results indicate that a hydrophobic substituent at the C-2 position plays a critical role in the appearance of biological activities, as in the case of substitution at C-9.
- Endo, Yasuyuki,Yokoyama, Akihiro
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- Powdered cosmetic compositions containing silicone elastomers
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A cosmetic powder is provided which includes a crosslinked non-emulsifying siloxane elastomer, a powdered inorganic material and a skin treatment agent. Inclusion of the elastomer allows for the coupling of water as well as water soluble salts into the cosmetic powder.
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- α hydroxylation of carboxylic acids with molecular oxygen catalyzed by the α oxidase of peas (Pisum sativum): A novel biocatalytic synthesis of enantiomerically pure (R)-2-hydroxy acids
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The substrate selectivities of the α oxidation of saturated, unsaturated, and heteroatom-containing (oxygen, sulfur) carboxylic acids 1 by the enzyme extract of peas (Pisum sativum) indicate that this biotransformation proceeds highly enantioselectively. For the first time, the synthesis of optically pure 2-hydroxy acids 2 has been achieved on the semipreparative scale (1 mmol) by α hydroxylation of long-chain carboxylic acids with molecular oxygen, catalyzed by the α oxidase of peas. For derivatives with sulfur atom in the chain, no sulfoxidation is observed. The functionalities (carbon double and triple bonds, oxygen, and sulfur atoms) must be at least three carbon atoms away from the carboxylic acid group to achieve efficient asymmetric hydroxylation. The absolute configuration of the 2-hydroxy acids 2 was assigned by comparison of the gaschromatographic data with that of authentic reference compounds and by application of the exciton- coupled-circular-dichroism (ECCD) method. This unprecedented asymmetric biocatalytic methodology should be valuable for the preparation of enantiomerically pure (R)-2-hydroxy acids.
- Adam, Waldemar,Boland, Wilhelm,Hartmann-Schreier, Jenny,Humpf, Hans-Ulrich,Lazarus, Michael,Saffert, Alexander,Saha-M?ller, Chantu R.,Schreier, Peter
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p. 11044 - 11048
(2007/10/03)
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- Anhydrous cosmetic compositions
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A skin treatment composition is provided which includes a crosslinked non-emulsifying siloxane elastomer, a skin conditioning agent, a volatile siloxane and no more than 5% water. Hydrocarbons such as petrolatum and polyols such as glycerin are the preferred skin conditioning agents. Inclusion of the elastomer provides a non-traditional smooth/silky feel to the skin upon application with a non-draggy rub in.
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- Anhydrous cosmetic composition with ceramides for firming skin
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A method and cosmetic composition for improving skin firmness are provided through an anhydrous composition including a hydrophobic carrier which may be a silicone or hydrocarbon for delivering an effective amount of a ceramide formed of a sphingoid base linked through an amide to a 2-hydroxycarboxylic C2-C30group.
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- Cerebrosides from Fomitopsis pinicola (Sw. Ex Fr.) Karst.
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A cerebroside fraction was obtained from the bruit bodies of fomitopsis pinicola using column chromatography and then separated into six compounds by reversed-phase HPLC.The sugar component of all cerebrosides was D-glucose.The major fatty acids were 2-hydroxyfatty acids (C14-C18), the long chain base was identified as 9-methyl-C18-4,8-sphingadienine which is widely distributed in fungi and reported to be essential for the fruit-inducing activity of fungi.Based on degradation studies, fast atom bombardment mass spectrometry, and different 1H and 13C NMR investigations, the structure of the main cerebroside (1) was determined to be (4E,8E,2S,3R,2'R)-N-2'-hydroxypalmityl-1-O-β-D-glucopyranosyl-9-methyl-4,8-sphingadienine. - Keywords: Cerebroside; Glycolipid; Fomitopsis pinicola; 1H and 13C NMR
- Striegler, S.,Haslinger, E.
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p. 755 - 762
(2007/10/03)
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- Enantioselective α hydroxylation of carboxylic acids with molecular oxygen catalyzed by the α oxidation enzyme system of young pea leaves (Pisum sativum): A substrate selectivity study
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The substrate selectivity of the α oxidation of carboxylic acids 1 by crude homogenate of young pea leaves was investigated. Saturated fatty acids with 7 to 16 carbon atoms and oleic acid were transformed to the enantiomerically pure (R)-2-hydroxy acids 2 in the presence of molecular oxygen.
- Adam, Waldemar,Lazarus, Michael,Saha-Moeller, Chantu R.,Schreier, Peter
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p. 2287 - 2292
(2007/10/03)
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- Cosmetic compositions for reducing or preventing signs of cellulite
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The invention is directed to increasing the strength and firmness of the skin and reducing the signs of cellulite. The inventive method includes applying to the skin a composition that includes inositol phosphate, particularly phytic acid and its salts, in a cosmetically acceptable carrier.
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- New potential immunoenhancing compounds. Synthesis and pharmacological evaluation of new long-chain 2-amido-2-deoxy-D-glucose derivatives
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A series of long-chain fatty acids and the corresponding 2-hydroxy, 2-oxo, 3-hydroxy acid glucosamides were evaluated as immunomodulating compounds. In a preliminary screening, 2-[(2-ethoxycarbonyloxy)tetradecanoyl-amino]-2-deoxy-D-glucose (2b) and 2-(3-hydroxydodecanoyl-amino)-2-deoxy-D-glucose (5a) resulted to be the most effective in enhancing the glucosamine activity. The findings of in vitro-ex vivo tests (unidirectional mixed lymphocyte culture reaction and primary antibody production) and in vivo tests (delayed type hypersensitivity, protection against bacterial or fungal infection and against Sarcoma 180 or Lewis lung carcinoma transplants) were very encouraging and allowed to assume for the two substances a protective activity, presumably through the ability of activating phagocytic and NK cells.
- Valcavi,Albertoni,Brandt,Corsi,Farina,Foresta,Pascucci,Ramacci
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p. 1190 - 1195
(2007/10/02)
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- Studies on angiotensin-converting enzyme inhibitors. I. Syntheses and angiotensin-converting enzyme inhibitory activity of 2-(3-merecaptopropionyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid derivatives
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(3S)-2-[(2S)-mercapto-2-methylpropionyl]-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid [(3S), (2S)-6a] was prepared by the reaction of (3S)-1,2,3,4-tetrahydreoisoquinoline-3-carboxylic acid test-butyl ester [(3S)-2a) or benzyl ester [(3S)-2b] with 3-benzoylthio-2-methylpropionyl chloride (3a), followed by fractional crystallization and removal of the protective group. The absolute configuration of (3S), (2S)-6a was confirmed by X-ray diffraction analysis of the thiazepinol[4,3-b]isoquinoline compound (7) derived from 6a. Resolution of 3-benzoylthio-2-methylpropionic acid (8) was completed by using optically active phenylalanine amide as a resolving agent. The other optical isomers of (3S),(2S)-6a were prepared by the reaction of (3S)- or (3R)-2b with optically active 3a. The in vitro ACE inhibitory activity of each isomer of 6a was evaluated. Among them, (3S),(2S)-6a was found to be the most potent inhibitor with an IC50 value of 8.6X10-9M. Compound (3S),(2S)-6a induced a dose-dependent inhibition of the pressor response to angiotensin 1 after oral administration to normotensive anesthetized rats. Moreover, (3S),(2S)-6a markedly reduced the systolic blood pressure in renal hypertensive rats (RHR) and spontaneously hypertensive rats (SHR). The in vivo ACE inhibitory activity and the hypotensive effects of (3S),(2S)-6a were comparable to those of captopril.
- Hayashi,Ozaki,Nunami,Uchida,Kato,Kinashi,Yoneda
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p. 570 - 576
(2007/10/02)
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