COSMETIC COMPOSITION COMPRISING A GRAFTED POLYHYDROXYALKANOATE COPOLYMER IN A FATTY MEDIUM

Abstract

The present invention relates to a cosmetic composition comprising a) one or more polyhydroxyalkanoate (PH A) copolymers which contain, and preferably consist of, at least two different repeating polymer units chosen from the units (A) and (B) below, and also the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof such as hydrates: -[-0-CH(R.sup.1)—CH.sub.2—C(0)-]- unit (A) -[-0-CH(R.sup.2)—CH.sub.2—C(0)-]- unit (B) in which polymer units (A) and (B): —R.sup.1 represents a hydrocarbon-based chain chosen from i) linear or branched (C.sub.5-C.sub.28)alkyl, ii) linear or branched (C.sub.6-C.sub.28)alkenyl, iii) linear or branched (C.sub.6-C.sub.28)alkynyl; preferably, the hydrocarbon-based group is linear; said hydrocarbon-based chain being optionally substituted and/or interrupted with atoms or groups as described in the description; —R.sup.2 represents a cyclic or non-cyclic, linear or branched, saturated or unsaturated hydrocarbon-based group, comprising from 3 to 30 carbon atoms; b) a fatty medium comprising one or more fatty substances which are preferably liquid at 25° C. and at atmospheric pressure; it being understood that (A) is different from (B).

Claims

1. The composition comprising: a) one or more polyhydroxyalkanoate (PHA) copolymers which contain at least two different repeating polymer units chosen from the units (A) and (B) below, and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof:
—[—O—CH(R.sup.1)—CH.sub.2—C(O)—]—  unit (A)
—[—O—CH(R.sup.2)—CH.sub.2—C(O)—]—  unit (B) in which polymer units (A) and (B): R.sup.1 represents a hydrocarbon-based chain chosen from i) linear or branched (C.sub.5-C.sub.28)alkyl, ii) linear or branched (C.sub.6-C.sub.28)alkenyl, iii) linear or branched (C.sub.6-C.sub.28)alkynyl; said hydrocarbon-based chain being: substituted with one or more atoms or groups chosen from: a) halogen, b) hydroxyl, c) thiol, d) (di)(C.sub.1-C.sub.4)(alkyl)amino, e) (thio)carboxy, f) (thio)carboxamide —C(O)—N(R.sub.a).sub.2 or C(S)—N(R.sub.a).sub.2, f) cyano, g) iso(thio)cyanate, h) (hetero)aryl, and i) (hetero)cycloalkyl, j) cosmetic active agent chosen from coloured or uncoloured, fluorescent or non-fluorescent chromophores, anti-ageing active agents and fragrances; k) R—X with R representing a group chosen from α) cycloalkyl, β) heterocycloalkyl, γ) (hetero)aryl, δ) cosmetic active agent as defined previously and X representing a′) O, S, N(R.sub.a) or Si(R.sub.b)(R.sub.c), b′) S(O).sub.r, or (thio)carbonyl, c′) or combinations of a′) with b′); r being equal to 1 or 2, R.sub.a representing a hydrogen atom, or a (C.sub.1-C.sub.4)alkyl group or an aryl(C.sub.1-C.sub.4)alkyl group; R.sub.b and R.sub.c, which may be identical or different, represent a (C.sub.1-C.sub.4)alkyl or (C.sub.1-C.sub.4)alkoxy group; and/or interrupted with one or more heteroatoms a′), b′), c′) or combinations of a′) with b′); R.sup.2 represents a cyclic or non-cyclic, linear or branched, saturated or unsaturated hydrocarbon-based group comprising from 3 to 30 carbon atoms; and b) a fatty medium comprising one or more fatty substances; it being understood that (A) is different from (B).

2. The composition according to claim 1, in which the PHA copolymer(s) a) contain the repeating unit of formula (I), and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof: ##STR00050## in which formula (I): m and n are integers greater than or equal to 1.

3. The composition according to claim 1, in which the PHA copolymer(s) a) contain three different repeating polymer units (A), (B) and (C), and preferably consist of three different polymer units (A), (B) and (C), below, and also the optical or geometrical isomers thereof and the solvates thereof:
—[—O—CH(R.sup.1)—CH.sub.2—C(O)—]—  unit (A)
—[—O—CH(R.sup.2)—CH.sub.2—C(O)—]—  unit (B)
—[—O—CH(R.sup.3)—CH.sub.2—C(O)—]—  unit (C) in which polymer units (A), (B) and (C): R.sup.3 represents a cyclic or non-cyclic, linear or branched, saturated or unsaturated hydrocarbon-based group comprising from 1 to 30 carbon atoms; optionally substituted with one or more atoms or groups a) to l) and/or optionally interrupted with one or more heteroatoms or groups a′) to c′); and it being understood that: (A) is different from (B) and (C), (B) is different from (A) and (C), and (C) is different from (A) and (B); and.

4. The composition according to claim 1, in which the PHA copolymer(s) a) contain four different repeating polymer units (A), (B), (C) and (D), below, and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof:
—[—O—CH(R.sup.1)—CH.sub.2—C(O)—]—  unit (A)
—[—O—CH(R.sup.2)—CH.sub.2—C(O)—]—  unit (B)
—[—O—CH(R.sup.3)—CH.sub.2—C(O)—]—  unit (C)
—[—O—CH(R.sup.4)—CH.sub.2—C(O)—]—  unit (D) in which polymer units (A), (B), (C) and (D): R.sup.4 represents a cyclic or non-cyclic, linear or branched, saturated hydrocarbon-based group comprising from 3 to 30 carbon atoms, optionally substituted with one or more atoms or groups a) to l) and/or optionally interrupted with one or more heteroatoms or groups a′) to c′); and it being understood that: (A) is different from (B), (C) and (D), (B) is different from (A), (C) and (D), (C) is different from (A), (B) and (D), and (D) is different from (A), (B) and (C).

5. The composition according to claim 1, in which the PHA copolymer(s) a) contain five different repeating polymer units (A), (B), (C), (D) and (E), below, and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof:
—[—O—CH(R.sup.1)—CH.sub.2—C(O)—]—  unit (A)
—[—O—CH(R.sup.2)—CH.sub.2—C(O)—]—  unit (B)
—[—O—CH(R.sup.3)—CH.sub.2—C(O)—]—  unit (C)
—[—O—CH(R.sup.4)—CH.sub.2—C(O)—]—  unit (D)
—[—O—CH(R.sup.5)—CH.sub.2—C(O)—]—  unit (E) in which polymer units (A), (B), (C), (D) and (E): R.sup.5 represents a cyclic or non-cyclic, linear or branched, saturated hydrocarbon-based group comprising from 3 to 30 carbon atoms, optionally substituted with one or more atoms or groups a) to l) and/or optionally interrupted with one or more heteroatoms or groups a′) to c′); it being understood that: (A) is different from (B), (C), (D) and (E); (B) is different from (A), (C), (D) and (E); (C) is different from (A), (B), (D) and (E); (D) is different from (A), (B), (C) and (E); and (E) is different from (A), (B), (C) and (D).

6. The composition according to claim 1, in which R.sup.1 represents a linear or branched (C.sub.5-C.sub.28)alkyl hydrocarbon-based chain.

7. The composition according to claim 1, in which R.sup.1 represents a hydrocarbon-based chain which is interrupted with one or more atoms or groups chosen from O, S, N(R.sub.a), carbonyl, or combinations thereof.

8. The composition according to claim 1, in which R.sup.1 has the following formula —(CH.sub.2).sub.r—X-(ALK).sub.u-G, ALK represents a linear or branched (C.sub.1-C.sub.10)alkylene chain, r represents an integer inclusively between 6 and 11; u is equal to 0 or 1; and G represents a hydrogen atom or a group chosen from hydroxyl, carboxyl, (di)(C.sub.1-C.sub.4)(alkyl)amino, (hetero)aryl, cycloalkyl, or a sugar.

9. The composition according to claim 1, in which the PHA copolymer(s) a) are such that R.sup.2 is chosen from linear or branched (C.sub.1-C.sub.28)alkyl, and linear or branched (C.sub.2-C.sub.28)alkenyl.

10. The composition according to claim 1, in which the PHA copolymer(s) a) are such that the radical R.sup.2 is a linear or branched (C.sub.1-C.sub.8)alkyl.

11. The composition according to claim 1, in which the PHA copolymer(s) a) are such that: the unit (A) is present in a molar percentage ranging from 0.1% to 99%; and the unit (B) is present in a molar percentage ranging from 1% to 40%; and/or the unit (C) is present in a molar percentage ranging from 0.5% to 20%.

12. (canceled)

13. The composition according to claim 1, in which the fatty medium comprises one or more substances chosen from: branched C.sub.8-C.sub.16 alkanes, linear C.sub.8-C.sub.16 alkanes; ester oils; esters of lactic acid and of C.sub.10-C.sub.20 alcohol; diesters of malic acid and of C.sub.10-C.sub.20 alcohol; esters of pentaerythritol and of C.sub.8-C.sub.22 carboxylic acid; diesters of formula R.sup.11—O—C(═O)—R.sup.12—C(═O)—O—R.sup.13, with R.sup.11 and R.sup.13, which may be identical or different, representing a linear or branched, saturated or unsaturated C.sub.4 to C.sub.12 alkyl chain, optionally containing at least one saturated or unsaturated, preferably saturated, ring, and R.sup.12 representing a saturated or unsaturated C.sub.1 to C.sub.4, alkylene chain; diesters of formula R.sup.14—C(═O)—O—R.sup.15—O—C(═O)—R.sup.16, with R.sup.14 and R.sup.16, which may be identical or different, representing a linear or branched, saturated or unsaturated C.sub.4 to C.sub.12 alkyl chain and R.sup.15 representing a saturated or unsaturated C.sub.1 to C.sub.4 alkylene chain; the carbonate oils may be chosen from the carbonates of the following formula R.sup.17—O—C(O)—O—R.sup.18, with R.sup.17 and R.sup.18, which may be identical or different, representing a linear or branched C.sub.4 to C.sub.12 alkyl chain; and mixtures thereof.

14. The composition according to claim 1, in which the fatty medium comprises one or more fatty substances in a content ranging from 2% to 99.9% by weight, relative to the total weight of the composition.

15. The composition according to claim 1, in which the fatty medium comprises one or more solvents.

16. The composition according to claim 1, which also comprises one or more colouring agents chosen from pigments, direct dyes and mixtures thereof.

17. A process for treating keratin materials by applying the composition as defined in claim 1.

18. A copolymer PHA which contain at least two different repeating polymer units chosen from the units (A) and (B) below, and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof:
—[—O—CH(R.sup.1)—CH.sub.2—C(O)—]—  unit (A)
—[—O—CH(R.sup.2)—CH.sub.2—C(O)—]—  unit (B) in which polymer units (A) and (B): R.sup.1 has the following formula —(CH.sub.2).sub.r—X-(ALK).sub.u-G, ALK represents a linear or branched-(C.sub.1-C.sub.10)alkylene chain, r represents an integer inclusively between 6 and 11 u is equal to 0 or 1; and G represents a hydrogen atom or a group chosen from hydroxyl, carboxyl, (di)(C.sub.1-C.sub.4)(alkyl)amino, (hetero)aryl, and R.sup.2 represents a cyclic or non-cyclic, linear or branched, saturated or unsaturated hydrocarbon-based group comprising from 3 to 30 carbon atoms; R.sup.1 has the following formula —(CH.sub.2).sub.r—X-(ALK).sub.u-G, ALK represents a linear or branched (C.sub.1-C.sub.10)alkylene chain, r represents an integer inclusively between 6 and 11; u is equal to 0 or 1; and G represents a hydrogen atom or a group chosen from hydroxyl, carboxyl, (di)(C.sub.1-C.sub.4)(alkyl)amino, (hetero)aryl.

19. A process for preparing the copolymer PHA as defined in claim 18, starting from a PHA copolymer selected from a) to g): a) a PHA copolymer bearing an unsaturated hydrocarbon-based chain, according to Scheme 1 below: ##STR00051## in which Scheme 1: m and n are integers greater than or equal to 1; Y represents a group chosen from Hal (halide), hydroxyl, thiol, (di)(C.sub.1-C.sub.4)(alkyl)amino, R—X with R representing a group chosen from α) cycloalkyl β) heterocycloalkyl, γ) (hetero)aryl; δ) a cosmetic active agent as defined previously; ε) (C.sub.1-C.sub.20)alkyl, (C.sub.2-C.sub.20)alkenyl, (C.sub.2-C.sub.20)alkynyl; and X representing a′) O, S, N(R.sub.a) or Si(R.sub.b)(R.sub.c) or e) linear or branched (C.sub.1-C.sub.20)alkyl, with R.sub.a, R.sub.b and R.sub.c as defined previously; q′ represents an integer inclusively between 2 and 20; A) the copolymer PHA with unsaturations may optionally be chemically modified: via addition reactions; B) the copolymer PHA with unsaturations may optionally be chemically modified via oxidation reactions, which may or may not be controlled; b) a PHA copolymer bearing a hydrocarbon-based chain containing an epoxide group, according to Scheme 2 below: ##STR00052## in which Scheme 2, Y, m, n, q′ and R.sup.2 are as defined in Scheme 1; c) a PHA copolymer bearing a hydrocarbon-based chain containing a nucleofugal group, according to Scheme 3 below: ##STR00053## in which Scheme 3 Y, m, n, q′ and R.sup.2 are as defined in Scheme 1; M corresponds to an organic or inorganic nucleofugal group, which may be substituted with a nucleophilic group; d) a PHA copolymer bearing a hydrocarbon-based chain containing a cyano group, according to Scheme 4 below: ##STR00054## in which Scheme 4 Y, m, n, q′ and R2 are as defined in Scheme 1; In a first step i), the PHA copolymer bearing a side chain containing a cyano or nitrile group reacts with an organo-alkali metal or organomagnesium compound Y-MgHal, Y—Li or Y—Na, followed by hydrolysis to give the PHA copolymer bearing a side chain containing a group Y grafted with a ketone function; the ketone function may be converted into a thio ketone by thionation; said thio ketone, after total reduction ii) leads to the PHA copolymer bearing a side chain containing a group Y grafted with an alkylene group; alternatively, said thio ketone may undergo a controlled reduction iii) with a conventional reducing agent to give the PHA copolymer bearing a side chain containing a group Y grafted with a hydroxyalkylene group; the cyano group of the starting PHA copolymer can react with water after hydration v) to give the amide derivative, or after hydrolysis iv) to the carboxyl derivative; the cyano group of the starting PHA copolymer may also, after reduction vi), give the amine derivative or the ketone derivative; e) a PHA copolymer bearing a hydrocarbon-based chain at the chain end, according to Scheme 5 below: ##STR00055## in which Scheme 5 R.sup.1, R.sup.2, m, n and Y are as defined previously, and R′.sup.1 represents a hydrocarbon-based chain chosen from i) linear or branched (C.sub.1-C.sub.20)alkyl, ii) linear or branched (C.sub.2-C.sub.20)alkenyl, iii) linear or branched (C.sub.2-C.sub.20)alkynyl; said hydrocarbon-based chain being substituted with one or more atoms or groups chosen from: a) halogens, b) hydroxyl, c) thiol, d) (di)(C.sub.1-C.sub.4)(alkyl)amino, e) (thio)carboxyl, f) (thio)carboxamide —C(O)—N(R.sub.a).sub.2 or —C(S)—N(R.sub.a).sub.2, f) cyano, g) iso(thio)cyanate, h) (hetero)aryl, and i) (hetero)cycloalkyl, j) a cosmetic active agent chosen from coloured or uncoloured, fluorescent or non-fluorescent chromophores, and anti-ageing active agents; or f) a PHA copolymers with reactive atom or group according to Scheme 6: ##STR00056## in which Scheme 6 R′.sup.1, R.sup.2, m, n and Y are as defined previously, and X′ represents a reactive atom or group that is capable of reacting with an electrophilic E or nucleophilic Nu atom or group to create a Σ covalent bond; if X′ is an electrophilic or nucleofugal group, then it can react with a reagent R′.sup.1—Nu; if X′ is a nucleophilic group Nu, then it can react with R′.sup.1-E to create a Σ covalent bond; the Σ covalent bonds or bonding group that may be generated are listed in the table below, from condensation of electrophiles with nucleophiles: TABLE-US-00018 Electrophiles  Nucleophiles   u Covalent bonds Activated esters* Amines Carboxamides Acyl azides** Amines Carboxamides Acyl halides Amines Carboxamides Acyl halides Alcohols Esters Acyl cyanides Alcohols Esters Acyl cyanides Amines Carboxamides Alkyl halides Amines Alkylamines Alkyl halides Carboxylic acids Esters Alkyl halides Thiols Thioesters Alkyl halides Alcohols Ethers Sulfonic acids and salt thereof Thiols Thioethers Sulfonic acids and salt thereof Carboxylic acids Esters Sulfonic acids and salt thereof Alcohols Ethers Anhydrides Alcohols Esters Anhydrides Amines Carboxamides Aryl halides Thiols Thioethers Aryl halides Amines Arylamines Azinridines Thiols Thioethers Carboxylic acids Amines Carboxamides Carboxylic acids Alcohols Esters Carbodiimides Carboxylic acids N-acylureas Diazolakanes Carboxylic acids Esters Epoxides Thiols Thioethers Haloacetamides Thiols Thioethers Imide esters Amines Amidines Isocyanates Amines Ureas Isocyanates Alcohols Urethanes Isothiocyanates Amines Thioureas Maleimides Thiols Thioethers Sulfonic esters Amines Alkylamines Sulfonic esters Thiols Thioethers Sulfonic esters Carboxylic acids Esters Sulfonic esters Alcohols Ethers Sulfonyl halides Amines Sulfonamides *the activated esters of general formula -CO-LG, with LG representing a leaving group such as oxysuccinimidyl, oxybenzotriazolyl or aryloxy, optionally substituted; **the acyl azides can rearrange to give isocyanate. g) a PHA functionalized on a side chain, to perform chain-end grafting in a second stage as described in Scheme 7; the reciprocal is also true, in which the chain-end grafting may be performed in a first stage, followed by performing functionalization of a functionalizable side chain in a second stage. ##STR00057## in which Scheme 7 R′.sup.1, R.sup.2, m, n and Y are as defined previously.

20. A cosmetic process which comprises applying the copolymer PHA as defined in claim 8 to human keratin fibres or human skin.

20. The composition according to claim 1, in which the PHA copolymer(s) a) are such that they comprise the following repeating units, and the optical or geometrical isomers thereof, the organic or mineral acid or base salts thereof, and the solvates thereof: ##STR00058## TABLE-US-00019 Compounds R.sup.1 R.sup.2 (1) -(CH.sub.2).sub.8-S-CH(CH.sub.3)-C(O)-OH -(CH.sub.2).sub.4-CH.sub.3 (2) -(CH.sub.2).sub.8-S-(CH.sub.2).sub.7-CH2 -(CH.sub.2).sub.4-CH.sub.3 (3) -(CH.sub.2).sub.8-S-(CH.sub.2).sub.8-OH -(CH.sub.2).sub.4-CH.sub.3 (4) -(CH.sub.2).sub.8-S-(CH2).sub.2-NH.sub.2 -(CH.sub.2).sub.4-CH.sub.3 (5) -(CH.sub.2).sub.8-S-Cycl -(CH.sub.2).sub.4-CH.sub.3 (6) -(CH.sub.2).sub.8-S-CH.sub.2-Fur -(CH.sub.2).sub.4-CH.sub.3 (7) -(CH.sub.2).sub.8-S-Sug -(CH.sub.2).sub.4-CH.sub.3 (8) -(CH.sub.2).sub.8-S-(CH.sub.2).sub.2-Ar -(CH.sub.2).sub.4-CH.sub.3 (9) -(CH.sub.2).sub.8-S-CH.sub.2-Ar -(CH.sub.2).sub.4-CH.sub.3 (10) -(CH.sub.2).sub.8-S-CH(CH.sub.3)-C(O)-OH -(CH.sub.2).sub.8-CH.sub.3 (11) -(CH.sub.2).sub.8-Hal -(CH.sub.2).sub.8-CH.sub.3 (12) -(CH.sub.2).sub.3-CN -(CH.sub.2).sub.8-CH.sub.3 (13) -(CH.sub.2).sub.8-CH.sub.3 (14) -(CH.sub.2).sub.2-Ar -(CH.sub.2).sub.8-CH.sub.3 m and n are integers greater than or equal to 1, Hal represents a halogen atom and t represents an integer between 1 and 10, Ar: represents a (hetero)aryl group; Ar′: represents a (C.sub.1-C.sub.4)alkyl(hetero)aryl group; Cycl: represents a cyclohexyl group; Fur: represents a furyl group; Sug: represents a sugar group.

Description

EXAMPLES

[0573] The PHAs illustrated in the various examples were prepared in 3-litre chemostats and/or 5-litre Fernbach flasks depending on whether or not a p-oxidation pathway inhibitor is used. The isolation of the PHAs is similar for all the examples obtained.

[0574] In a first step, the microorganism generates the PHAs which are stored in intracellular granules, the proportion of which varies as a function of the applied conditions such as the temperature or the nature of the culture medium. The generation of PHA granules may or may not be associated with the growth of the microorganism as a function of the nature of the microorganisms. During the second step, the biomass containing the PHAs is isolated, i.e. separated from the fermentation medium, and then dried. The PHAs are extracted from the biomass before being purified, if necessary.

[0575] A mixture of saturated and unsaturated carbon sources is, for certain examples, necessary for the stability of the PHA obtained.

TABLE-US-00008 TABLE 2 Carbon source CAS Caprylic add (RADIACID 608) 124-07-2 Nonanoic acid 112-05-0 Undecylenic acid (10-Undecencic acid) 112-38-9

TABLE-US-00009 TABLE 3 Carbon source Genus and species Source Caprylic and undecylenoic Pseudomonas ATCC ® 47054 ™ acid mixture putida Nonanoic and undecylenoic Pseudomonas ATCC ® 47054 ™ acid mixture putida

Example 1: PHA Bearing a Side Chain R.SUP.1 .Representing a Linear 10% Unsaturated n-octenyl Group and R.SUP.2 .Representing an n-Pentyl Group

[0576] ##STR00030##

[0577] The process for synthesizing the compound of Example 1 is adapted from the article: Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440, Z. Sun, J. A. Ramsay, M. Guay, B. A. Ramsay, Applied Microbiology Biotechnology, 82, 657-662, 2009.

[0578] The microorganism used is Pseudomonas putida KT2440 ATCC® 47054™. The culture method is performed under fed-batch growth axenic conditions with a maintenance solution containing a mixture of carbon source at a rate μ=0.15 h.sup.−1 in a 3 L chemostat containing 2.5 L of culture medium.

[0579] The system is aerated with a flow of 0.5 vvm of air for a nominal dissolved oxygen (O.sub.D) value at 30% of saturation. The pH is regulated with 15% aqueous ammonia solution. The temperature of the fermentation medium is regulated at 30° C.

[0580] Assembly for the Fed-Batch Growth Fermentation Mode

[0581] The fermentation medium is regulated in terms of temperature-pressure of dissolved oxygen and pH (not shown)

[0582] See FIG. 1

[0583] The production process is performed using three different culture media. The first culture medium, defined CM1 “inoculum”, is used for the preparation of the preculture. The second culture medium, defined CM2 “batch”, is used for unfed batch growth of the microorganism with the primary carbon sources in the Fernbach flasks. The third culture medium, defined CM3 “maintenance”, is used for the fed-batch or maintenance fermentation mode with the carbon sources of interest at a flow rate calibrated as a function of the growth of the microorganism.

[0584] Composition in Grams Per Litre

TABLE-US-00010 TABLE 4 CM1 CM2 CM3 Ingredients « incculum » « batch » «maintenance » (NH.sub.4).sub.2SO.sub.4 4.7 4.7 Na.sub.2HPO.sub.4•7H.sub.2O 12 9 KH.sub.2PO.sub.4 2.7 2.03 MgSO.sub.4•7H.sub.2O 0.8 1.03 Nutrient Broth 3 / Caprylic acid / 0.9 900 Undecylenic acid / 0.1 100 Microelement solution / 10 2N NaOH QSP pH = 6.8 MilliQ water QSP 1000 g

[0585] The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.

[0586] Composition of the Solution of Microelements in Grams Per Litre:

TABLE-US-00011 TABLE 5 Ingredients Amount FeSO.sub.4•7H.sub.2O 10.0 g CaCl.sub.2•2H.sub.2O 3.0 g ZnSO.sub.4•7H.sub.2O 2.2 g MnSO.sub.4•4H.sub.2O 0.5 g H.sub.3BO.sub.3 0.3 g CoCl.sub.2•6H.sub.2O 0.2 g Na.sub.2MoO.sub.4•2H.sub.2O 0.15 g NiCl.sub.2•6H.sub.2O 0.02 g CuSo.sub.4•5H.sub.2O 1.00 g MilliQ water (or 0.5N HCL) QSP 1000 g

[0587] 100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and are then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 3 L chemostat are inoculated at OD=0.1 with the 100 mL of preculture. After 4 hours at 30° C. at 850 rpm, the introduction of the maintenance medium is performed by applying the flow rate defined by equation 1.

[0588] At the end of the introduction, the biomass is isolated by centrifugation and then washed three times with water. The biomass is dried by lyophilization before being extracted with ethyl acetate for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, the PHA compound dissolved in the ethyl acetate, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.

[0589] The PHA may optionally be purified by successive dissolution and precipitation from an ethyl acetate/ethanol 70% methanol system, for example.

[0590] The PHA was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.

Example 2: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 100% Grafted with Thiolactic Acid (Compound of Example 1 Grafted with Thiolactic Acid TLA)

[0591] ##STR00031##

[0592] 1 g of the compound of Example 1 and 150 mg of thiolactic acid were dissolved in 20 mL of ethyl acetate at room temperature with stirring. 20 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0593] 20 mL of the reaction medium were then precipitated from a 200 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0594] The grafted PHA of Example 2 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.

Example 3: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 100% Grafted with Octanethiol (Compound of Example 1 Grafted with n-octanethiol)

[0595] ##STR00032##

[0596] 0.5 g of the compound of Example 1 and 125 mg of octanethiol were dissolved in 10 mL of ethyl acetate at room temperature with stirring. 15 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0597] The reaction medium was then precipitated from a 100 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0598] The grafted PHA of Example 3 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.

Example 4: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 75% Grafted with 8-mercapto-1-octanol (Compound of Example 1 Grafted with 8-mercapto-1-octanol)

[0599] ##STR00033##

[0600] 50 mg of the compound of Example 1 and 10 mg of 8-mercapto-1-octanol were dissolved in 5 mL of ethyl acetate at room temperature with stirring. 2 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0601] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0602] The grafted PHA of Example 4 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 75% or 7.5% of functions in total.

Example 5: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 32% Grafted with Cysteamine (Compound of Example 1 Grafted with Cysteamine)

[0603] ##STR00034##

[0604] 0.5 g of the compound of Example 1 and 54 mg of cysteamine were dissolved in a mixture of 10 mL of dichloromethane and 2 mL of ethanol at room temperature with stirring. 10 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0605] The reaction medium was then precipitated from a 100 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0606] The grafted PHA of Example 5 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 32% (see the spectrum below) or 3.2% of functions in total.

Example 6: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 73% Grafted with Cyclohexanethiol (Compound of Example 1 Grafted with CHT)

[0607] ##STR00035##

[0608] 100 mg of the compound of Example 1 and 26 mg of cyclohexanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0609] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0610] The grafted PHA of Example 6 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 73% or 7.3% of functions in total.

Example 7: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 66% Grafted with 2-furanmethanethiol (FT) (Compound of Example 1 Grafted with FT)

[0611] ##STR00036##

[0612] 100 mg of the compound of Example 1 and 26 mg of 2-furanmethanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0613] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0614] The grafted PHA of Example 7 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 66% or 6.6% of functions in total.

Example 8: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 66% Grafted with 1-thio-β-D-glucose tetraacetate (Compound of Example 1 Grafted with TGT)

[0615] ##STR00037##

[0616] 100 mg of the compound of Example 1 and 26 mg of 1-thio-β-D-glucose tetraacetate were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0617] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0618] The grafted PHA of Example 8 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 70% or 7% of functions in total.

Example 9: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 50% Grafted with 2-phenylethanethiol (PT) (Compound of Example 1 Grafted with PT)

[0619] ##STR00038##

[0620] 100 mg of the compound of Example 1 and 26 mg of 2-phenylethanethiol were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0621] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0622] The grafted PHA of Example 9 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 50% or 5% of functions in total.

Example 10: Poly(3-hydroxyoctanoate-co-undecenoate) Containing 10% Unsaturations 64% Grafted with 4-tert-butylbenzyl mercaptan (TBM) (Compound of Example 1 Grafted with TBM)

[0623] ##STR00039##

[0624] 100 mg of the compound of Example 1 and 26 mg of 4-tert-butylbenzyl mercaptan were dissolved in 5 mL of dichloromethane at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0625] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0626] The grafted PHA of Example 10 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 64% or 6.4% of functions in total.

Example 11: Poly(3-hydroxynonanoate-co-undecenoate) Containing 10% Unsaturations 100% Grafted with Thiolactic Acid

[0627] ##STR00040##

[0628] 0.1 g of the compound of Example 1 and 15 mg of thiolactic acid were dissolved in 5 mL of chloroform at room temperature with stirring. 5 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0629] The reaction medium was then precipitated from a 50 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0630] The grafted PHA of Example 11 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.

Example 12: Poly(3-hydroxynonanoate-co-undecenoate) Containing 5% Unsaturations 100% Grafted with Octanethiol

[0631] ##STR00041##

Preparation of Example 1′: Copolymer of PHA Bearing a Side Chain R.SUP.1 .Representing an n-hexyl Group and R.SUP.2 .Representing an n-hexyl Group

[0632] ##STR00042##

[0633] The production process of Example 1 is adapted to that of Example 1′, replacing the n-octanoic acid carbon source of Example 1 with n-nonanoic acid.

[0634] The PHA copolymer of Example 1′ was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure, with a degree of unsaturation of 5%.

[0635] 1 g of the PHA copolymer of Example 1′ and 150 mg of octanethiol were dissolved in 15 mL of ethyl acetate at room temperature with stirring. 20 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0636] The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0637] The grafted PHA of Example 12 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.

Example 13: Poly(3-hydroxynonanoate-co-undecenoate) Containing 5% Unsaturations 100% Epoxidized

[0638] ##STR00043##

[0639] 20 g of the PHA copolymer of Example 1′ were dissolved in 80 mL of anhydrous dichloromethane. A suspension of 1.9 g of 77% m-CPBA was prepared with 20 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.

[0640] The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0641] The PHA of Example 13 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.

Example 14: Poly(3-hydroxynonanoate-co-undecenoate) Containing 10% Unsaturations 100% Epoxidized

[0642] 10 g of the PHA copolymer identical to that of Example 1′ but with a degree of unsaturation of 10% were dissolved in 40 mL of anhydrous dichloromethane. A suspension of 1.9 g of 77% m-CPBA was prepared with 10 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.

[0643] The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0644] The PHA of Example 14 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.

Example 15: Poly(3-hydroxynonanoate-co-undecenoate) Containing 30% Unsaturations 100% Epoxidized

[0645] 10 g of the PHA copolymer identical to that of Example 1′ but with a degree of unsaturation of 30% were dissolved in 40 mL of anhydrous dichloromethane. A suspension of 6.2 g of 77% m-CPBA was prepared with 10 mL of anhydrous dichloromethane and added to the mixture with stirring, at room temperature for at least 120 hours.

[0646] The reaction medium was then precipitated from a 250 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0647] The PHA of Example 15 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Epoxidation to 100%.

Example 16: Poly(3-hydroxynonanoate-co-undecenoate) Containing 5% Unsaturations 100% Grafted with 4-tert-butylbenzyl Mercaptan (TBM) (Compound of Example 1′ Grafted with TBM)

[0648] ##STR00044##

[0649] 2 g of the PHA copolymer of Example 1′ and 300 mg of 4-tert-butylbenzyl mercaptan were dissolved in 25 mL of ethyl acetate at room temperature with stirring. 25 mg of 2,2-dimethoxy-2-phenylacetophenone (Irgacure 651) were added to the mixture. The medium was then irradiated under a 100 W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0650] The reaction medium was then precipitated from a 500 mL mixture of 70/30 v/v ethanol/water. A viscous white precipitate was obtained. This step may be repeated. The product thus obtained was dissolved in a minimum amount of ethyl acetate, poured onto a Teflon plate and then dried under dynamic vacuum at 40° C. to obtain a homogeneous film.

[0651] The PHA of Example 16 was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure. Grafting to 100%.

Example 17: PHA Bearing a Side Chain R.SUP.1 .Representing a 5% Linear 8-bromo-n-octanoyl Group and R.SUP.2 .Representing a n-hexyl Group

[0652] ##STR00045##

[0653] The process for synthesizing the compound of Example 1 is adapted from the article: Fed-batch production of unsaturated medium-chain-length polyhydroxyalkanoates with controlled composition by Pseudomonas putida KT2440, Z. Sun, J. A. Ramsay, M. Guay, B. A. Ramsay, Applied Microbiology Biotechnology, 82, 657-662, 2009.

[0654] The microorganism used is Pseudomonas putida KT2440 ATCC® 47054™. The culture method is performed under fed-batch growth axenic conditions with a maintenance solution containing a mixture of carbon source at a rate μ=0.15 h.sup.−1 in a 3 L chemostat containing 2.5 L of culture medium.

[0655] The system is aerated with a flow of 0.5 vvm of air for a dissolved oxygen (O.sub.D) value at 30% of saturation. The pH is regulated with a solution composed of ammonia and glucose with a final mass of respectively 15% and 40%. The temperature of the fermentation medium is regulated at 30° C.

[0656] Assembly for the Fed-Batch Growth Fermentation Mode:

[0657] The fermentation medium is regulated in terms of temperature-pressure of dissolved oxygen and pH (not shown)

[0658] See FIG. 1

[0659] The production process is performed using three different culture media. The first culture medium, defined CM1 “inoculum”, is used for the preparation of the preculture. The second culture medium, defined CM2 “batch”, is used for unfed batch growth of the microorganism with the primary carbon sources in the Fernbach flasks. The third culture medium defined CM3 “maintenance” is used for the batch, or maintenance, feeding of the fermentation with the carbon sources of interest at a flow rate calibrated as a function of the growth of the microorganism.

TABLE-US-00012 TABLE 6 CM1 CM2 CM3 Ingredients in grams per litre “inoculum” “batch” “maintenance” (NH.sub.4).sub.2SO.sub.4 4.7 4.7 Na.sub.2HPO.sub.4•7H.sub.2O 12 9 KH.sub.2PO.sub.4 2.7 2.03 MgSO.sub.4•7H.sub.2O 0.8 1.03 Nutrient broth 3 / Nonanoic acid / 1 923 11-Bromoundecanoic acid / 0 77 Microelement solution / 10 2N NaOH QSP pH = 6.8 MilliQ water QSP 1000 g

[0660] The composition of the Nutrient Broth, as mass percentages, is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™.

TABLE-US-00013 TABLE 7 Ingredients in grams per litre Amount FeSO.sub.4•7H.sub.2O 10.0 g CaCl.sub.2•2H.sub.2O 3.0 g ZnSO.sub.4•7H.sub.2O 2.2 g MnSO.sub.4•4H.sub.2O 0.5 g H.sub.3BO.sub.3 0.3 g CoCl.sub.2•6H.sub.2O 0.2 g Na.sub.2MoO.sub.4•2H.sub.2O 0.15 g NiCl.sub.2•6H.sub.2O 0.02 g CuSO.sub.4•5H.sub.2O 1.00 g MilliQ water (or 0.5N HCI) QSP 1000 g

[0661] 100 mL of preculture are prepared by suspending a cryotube containing 1 mL of the strain with 100 mL of “inoculum” culture medium at a pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask and then incubated at 30° C. at 150 rpm for 24 hours. 1.9 L of CM2 “batch” culture medium placed in a presterilized 3 L chemostat are inoculated at OD=0.1 with 100 mL of preculture. After 4 hours at 30° C. at 850 rpm, the introduction of the maintenance medium is performed by applying the flow rate defined by equation 1.

[0662] At the end of introduction, the biomass is isolated by centrifugation and then washed three times with water. The biomass is dried by lyophilization before being extracted with ethyl acetate for 24 hours. The suspension is clarified by filtration on a GF/A filter (Whatman®). The filtrate, composed of PHA dissolved in ethyl acetate, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass.

[0663] The PHA may optionally be purified by successive dissolution and precipitations in an ethyl acetate/ethanol 70% methanol system for instance.

[0664] The PHA was fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure: 95 molar % of units (B) for which R.sup.2=n-hexyl (71%) and n-butyl (24%) and 5 molar % of units (A) for which R.sup.1=8-bromo-n-octanyl (5.9%) and 6-bromo-n-hexyl (0.2%).

Evaluations

[0665] In a first stage, a film is prepared on a contrast card with a film spreader (speed: 50 mm/s-Cylinder: 100 μm). The film is left to dry for 24 hours at room temperature. Once dry, the film has a thickness of about 40 μm.

[0666] For the PHA copolymers of Examples 1 to 4 that are soluble in isododecane or an isododecane/ethanol mixture, evaluation of the cosmetic properties on a dry film was performed.

[0667] In a first stage, a film is prepared on a contrast card with a film spreader (speed: 50 mm/s-Cylinder: 100 μm). The film is left to dry for 24 hours at room temperature. Once dry, the film has a thickness of about 40 μm.

[0668] Three evaluations are performed on the dry film: Resistance to fats, gloss and tackiness

Measurement of the Resistance to Fats

[0669] Three drops of olive oil or sebum or water were deposited on the dry film present on the black part of the contrast card. Each drop corresponds to about 10 μL of olive oil (use of a micropipette).

[0670] The drop is left in contact with the dry film for two times: 5 minutes and 30 minutes. Once the time has elapsed, the drop of olive oil or sebum or water is wiped off and observation of the deterioration of the polymer film is performed. If the film was damaged by the drop of olive oil or sebum or water, the polymer film is regarded as being non-resistant to olive oil or to sebum.

[0671] Evaluations of the Polymers Only Soluble in Isododecane and Isododecane/Ethanol for Resistance to Water, Oil and Sebum:

TABLE-US-00014 TABLE 8 Tests Example 12 Example 13 Example 16 Example 17 Resistance to +++ +++ +++ +++ water Resistance to +++ +++ +++ +++ olive oil Resistance to ++ ++ ++ ++ sebum

[0672] It is seen that the PHA copolymers of the invention make it possible to obtain dry, homogeneous films that are particularly resistant to water, olive oil and sebum.

[0673] Measurement of the Gloss

[0674] Measurement of the gloss with a glossmeter on the black part of the contrast card. The gloss is read at an angle of 20° (the most discerning angle).

[0675] Evaluations of the Gloss on the Polymers Alone Soluble in Isododecane and Isododecane/Ethanol:

TABLE-US-00015 TABLE 9 Tests Example 12 Example 13 Example 16 Example 17 Gloss at Glossy (50) Not glossy (3) Glossy (70) Not glossy (3) 20°

[0676] The tack was evaluated in a sensory and qualitative manner by touching the dry film with a finger.

[0677] It is seen that Example 13 tested does not have a tacky feel.

Measure of the Resistance Vs Water/Oil and Adhesive Tape can Also be Evaluated

[0678] Mixing of the polymer dissolved in isododecane or isododecane/ethanol with the pigment for 2 minutes at 3500 rpm. The evaluations are performed on BioSkin. In a first stage, a film of each formulation is deposited on a BioSkin sample by means of a film spreader. The thickness of the wet film is 100 μm. The films are dried for 24 hours at room temperature. Once the films are dry, the tests may be performed.

Resistance to Olive Oil/Sebum

[0679] 0.5 mL of olive oil or sebum is applied to the film of formulation. After 5 minutes, the olive oil or sebum is removed by wiping 15 times with cotton wool. The deterioration of the film following contact with the olive oil or the sebum is thus examined (see FIG. 2).

Resistance to Adhesive Tapes

[0680] A strip of adhesive tape (of Scotch® type) is applied to the film of formulation. A weight is applied to the strip of said tape for 30 seconds. The adhesive tape is then removed and mounted on a slide holder so as to observe the result. The adherence of the film to the support is thus evaluated (see FIG. 2).

Example 18: Poly(3-HydroxyNonanoate-co-Undécenoate) with 5% (PHNUn5) and Grafted at 100% with 2-(Trimethylsilyl)ethanthiol (PHNUn5-g-TMS)

[0681] ##STR00046##

[0682] In order to synthesize the intermediate mc1-PHA with linear side chain R.sup.1═C.sub.8 alkenyle group and R.sup.2 n-hexyl with unsaturated at 5% (PHNUn5) the process is identical than the one discloses in Example 11.

[0683] Poly (3-HydroxyNonanoate-co-Undecenoate) is functionalized at 5% unsaturations with 2-(Trimethylsilyl) ethanthiol (PHNUn5 grafted 2-(Trimethylsilyl) ethanthiol) 1 g of (PHNUn5) and 300 mg of 2-(Trimethylsilyl) ethanthiol (TMS) are dissolved in 10 mL of ethyl acetate at room temperature with stirring. Then 20 mg of 2,2-Dimethoxy-2-phenylacetophenone (IRGACURE 651) is added to the mixture. The medium is then irradiated under a 100W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes.

[0684] The reaction medium is then precipitated from a 100 ml mixture of 70/30 vv ethanol/water. A viscous white precipitate is obtained. The latter step is repeated if necessary. The product thus obtained is dissolved in a minimum of ethyl acetate, poured onto a Teflon plate, then dried under dynamic vacuum at 40° C., to obtain a homogeneous film.

[0685] The PHA grafted with 2-(Trimethylsilyl) ethanthiol is fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure, 100% was grafted.

Example 19: Poly (3-HydroxyNonanoate-co-Undecenoate) with 1% Unsaturations Grafted at 100% with Thiolactic Acid (PHNUn1 Grafted with TLA)

[0686] ##STR00047##

Preparation of mc1-PHA with Linear Side Chain R.SUP.1.═C.SUB.8 .Alkenyl and R.SUP.2.=n-hexyl and Unsaturated at 1% (PHNUn1)

[0687] ##STR00048##

[0688] The process for obtaining PHNUn1 is adapted from the article: Applied Microbiololy Biotechnology, Z. Sun, et al., 82, 657-662. (2009).

[0689] The microorganism used is Pseudomonas putida KT2440 ATCC® 47054™. The culture mode is carried out under axenic conditions in discontinuous growth fed with a maintenance solution containing a mixture of carbon source at a rate of μ=0.15 h-1 in a 3 L chemostat containing 2.5 L of culture medium. The flow rate of the maintenance feed pump is proportional to the growth of the microorganism according to Equation 1:

[00001]$\mathrm{St}=\frac{X_t}{Y_{X/S}}=\frac{X_0}{Y_{X/S}}{e}^{\mathrm{.Math.}.t}$

[0690] The production process is carried out using three separate culture media. The first culture medium defined MC1 “inoculum” is used for the preparation of the preculture. The second culture medium defined MC2 “bach” is used for the non-fed discontinuous growth of the microorganism with the primary carbon sources in the Fernbachs flasks. The third culture medium defined MC3 “maintenance” is used for the batch feeding, or maintenance, of the fermentation with the carbon sources of interest at a rate calibrated according to the growth of the microorganism.

[0691] The composition in grams per liter of the three media is described in Table 10:

TABLE-US-00016 TABLE 10 MC1 MC2 MC3 Ingredients in grams per litre « inoculum » « batch » «maintenance » (NH.sub.4).sub.2SO.sub.4 4.7 4.7 Na.sub.2HPO.sub.4; 7H.sub.2O 12 9 KH.sub.2PO.sub.4 2.7 2.03 MgSO.sub.4; 7H.sub.2O 0.8 1.03 Nutrient Broth 3 / Nonanoic acid / 0.99 990 Undecylenic acid 0.01 10 Microelements Solution / 10 Acrylic acid / / NaOH 2N QSP pH = 6.8 MilliQ water QSP m = 1000 g

Composition in Grams Per Liter of Culture Media for Preculture and Maintenance.

[0692] The composition of Nutrient Broth in percentage by mass is 37.5% beef extract and 62.5% peptone. Reference 233000 DIFCO™. The composition of the solution of microelements in grams per liter is described in Table 7.

[0693] 100 mL of preculture is prepared by suspending a cryotube containing 1 mL of the strain with 100 mL “inoculum” culture media at pH adjusted to 6.8 with 2N NaOH in a 250 mL Fernbach flask then incubate at 30° Cat 150 rpm for 24 hours. 1.9 L of “BATCH” MC2 culture medium placed in a previously sterilized 3 L chemostat are inoculated at OD=0.1 with the 100 mL of preculture. After 4 hours at 30° C. at 850 rpm, the introduction of maintenance is performed by applying the flow rate defined by equation 1.

[0694] At the end of the introduction, the biomass is isolated by centrifugation and then washed three times with water. The biomass is dried by lyophilization before being extracted with ethyl acetate for 24 hours. The suspension is clarified by filtration through a GF/A filter (Wattman®), the filtrate, composed of PHA dissolved in ethyl acetate, is concentrated by evaporation and then dried under high vacuum at 40° C. to constant mass. The PHA can optionally be purified by solubilization and successive precipitations such as an ethyl acetate/Ethanol 70% methanol mixture.

[0695] Then 1 g of (PHNUn1) and 40 mg of thiolactic acid are dissolved in 10 mL of ethyl acetate at room temperature with stirring. 10 mg of 2,2-Dimethoxy-2-Phenylacetophenone (IRGACURE 651) is added to the mixture. The medium is then irradiated under a 100W UV lamp at 365 nm (reference) and with stirring for at least 10 minutes. The reaction medium is then precipitated from a 100 ml mixture of 70/30 vv ethanol/water. A viscous white precipitate was obtained. This step can be repeated. The product thus obtained is dissolved in a minimum of ethyl acetate, poured onto a Teflon plate, then dried under dynamic vacuum at 40° C., to obtain a homogeneous film.

[0696] PHA of example 19 grafted with thiolactic acid was characterized spectrometric method and show that the signals characteristic of the unsaturations have completely disappeared. 100% grafting.

Example 20: Poly (3-HydroxyNonanoate-Co-Undecenoate) at 1% Grafted with Thiolactic Acid Grafted with Dansylcadaverine (PHNUn1-g-TLA-g-Dansylcadaverine)

[0697] ##STR00049##

[0698] 1 g of the preceeding example 19, 12 mg of O-(1H-6-Chlorobenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU), 10 mg 4-methylmorpholine (NMM) and 12 mg of dansylcadaverine are dissolved in 10 mL of chloroform at room temperature with stirring for 24 h.

[0699] The reaction medium is then precipitated from a 100 ml mixture of 70/30 vv ethanol/water. A viscous, slightly yellow colorless precipitate is obtained. This step is repeated if necessary. The product thus obtained is dissolved in a minimum of ethyl acetate, poured onto a Teflon plate, then dried under dynamic vacuum at 40° C., to obtain a homogeneous film.

[0700] The PHA is fully characterized by spectroscopic and spectrometric methods and is in accordance with the expected chemical structure.

[0701] Evaluations of the Polymers Only Soluble in Isododecane and Isododecane/Ethanol for Resistance to Water, Oil and Sebum:

TABLE-US-00017 TABLE 10 Tests Example 18 Example 19 Example 20 Resistance to +++ +++ +++ water Resistance to +++ +++ +++ olive oil Resistance to ++ ++ ++ sebum

[0702] It is seen that the PHA copolymers of the invention make it possible to obtain dry, homogeneous films that are particularly resistant to water, olive oil and sebum.

[0703] Moreover PHA of the example 20 is evaluated for the resistance vs. water, and sebum under UV lamp. The latter experiment shows an intense and persistent flurescence of the film of example 20.