CH626403A5 - Process for the preparation of tertiary, optically active aliphatic compounds - Google Patents

Abstract

A microbiological process for the preparation of tertiary, optically active aliphatic compounds of the formula <IMAGE> is described. In this process, a microorganism which is an aerobe or facultative aerobe and which hydrogenates a double bond located between CH and methylated C in an aliphatic chain is allowed to act on a compound of the formula <IMAGE> in an aqueous medium. X, Y, X' and Y' in the formulae I and II have the meaning stated in Claim 1. The compounds of the formula II are intermediates in the synthesis of, inter alia, natural, optically active vitamin E, vitamin E esters and vitamin K1.

Description

       

  
 

** WARNING ** beginning of DESC field could overlap end of CLMS **. 

 



      PATENT REQUESTS
1.  Process for the preparation of tertiary optically active aliphatic compounds of the general formula
EMI1. 1
 in which one of the radicals X 'and Y' represents optionally esterified carboxy and the other represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl, where X 'and Y' have different functional meanings and furthermore free carboxy and hydroxymethyl groups with one another to form the group -CO-O-CH2- or -CH2-O-CO- are lactonized, and if X 'represents alkyl-etherified hydroxymethyl, Y' can also optionally be esterified hydroxymethyl, characterized in that an aerobic or facultative aerobic microorganism containing one in hydrogenates an aliphatic chain between CH and methylated C double bond,

   to a compound of the general formula
EMI1. 2nd
 in which X represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl and Y optionally esterified carboxy, optionally acetalized formyl or optionally etherified or esterified hydroxymethyl, where X and Y have different functional meanings and furthermore, if X is optionally esterified hydroxymethyl, Y is esterified Represents carboxy or optionally acetalized formyl and, if Y represents optionally esterified hydroxymethyl, X represents esterified carboxy or alkyl-etherified hydroxymethyl, can act in an aqueous medium.    



   2nd  A method according to claim 1, characterized in that one carries out the fermentative hydrogenation under aerobic conditions with a microorganism in a non-growing (stationary) phase. 



   3rd  A method according to claim 1 or 2, characterized in that one carries out the fermentative hydrogenation in the presence of an assimilable carbon source. 



   4th  A method according to claim 3, characterized in that a sugar in an amount of about 10 to 100 g per liter of aqueous medium is used as the carbon source. 



   5.  Method according to one of claims 1 to 4, characterized in that one carries out the fermentative hydrogenation at a pH of 2 to 10, in particular 3 to 8. 



   6.  Method according to one of claims 1 to 5, characterized in that one carries out the fermentative hydrogenation at a temperature of 20 to 35 C. 



   7.  Process according to one of claims 1 to 6, characterized in that the starting material of the formula I is used in a concentration of 0.1 to 5.0%, in particular 0.5 to 2.5%, based on the aqueous medium. 



   8th.  Process according to one of Claims 1 to 7, characterized in that a starting compound of the formula I is used in which X is optionally esterified carboxy and Y is optionally acetalized formyl or X is esterified carboxy and Y is optionally etherified or esterified hydroxymethyl, and as a microorganism Saccharomyces cerevisiae (Pressed yeast; baker's yeast) is used to obtain a fermentation product of the formula II in which X 'represents optionally esterified carboxy and Y / optionally etherified or esterified hydroxymethyl, free carboxy and hydroxymethyl groups being formed together to form the group -CO-O-CH2 - are lactonized. 



   9.  A method according to claim 8, characterized in that one uses a starting compound of formula I in which X represents optionally esterified carboxy and Y optionally acetalized formyl. 



   10th  Process according to Claim 9, characterized in that ethyl trans4,4-dimethoxy-3-methylcrotonate is used as the starting compound of the formula I, the main product being (S) -3 methyl-4-hydroxy-butyric acid ethyl ester as the fermentation product of the formula II which is converted into (S) -dihydro-4-methyl-2 (3H) -furanone by acid hydrolysis. 



   The invention relates to a process for the preparation of tertiary, optically active aliphatic compounds of the general formula
EMI1. 3rd
 in which one of the radicals X 'and Y' represents optionally esterified carboxy and the other represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl, where X 'and Y' have different functional meanings and furthermore free carboxy and hydroxymethyl groups with one another to form the group -COSCH2- or -CH2-OCO are lactonized, and if X 'represents alkyl-etherified hydroxymethyl, Y' can also optionally be esterified hydroxymethyl. 



   The process according to the invention is characterized in that an aerobic or facultative aerobic microorganism which hydrogenates a double bond located in an aliphatic chain between CH and methylated C, on a compound of the general formula
EMI1. 4th
 in which X represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl and Y optionally esterified carboxy, optionally acetalized formyl or optionally etherified or esterified hydroxymethyl, where X and Y have different functional meanings and furthermore, if X is optionally esterified hydroxymethyl, Y is esterified Represents carboxy or optionally acetalized formyl and if Y optionally represents esterified hydroxymethyl, X represents esterified carboxy or alkyl-etherified hydroxymethyl,

   can act in an aqueous medium.    



   The process according to the invention opens up a new advantageous route to intermediates in the synthesis of natural, optically active vitamin E, vitamin E ester and vitamin Kl, compounds which previously had to be obtained in an uneconomical manner from natural products or by means of complex synthesis methods.  Because of the new method according to the invention, new, optically active compounds of formula II are obtained in a particularly simple manner, which can be built up advantageously to the optically active vitamins mentioned.  Overall, the new route to natural, optically active vitamin E, vitamin E ester and vitamin Kl, like the method according to the invention itself, is therefore a valuable addition to the prior art. 



   The condition defined above that X and Y or  X 'and Y' have different functional meanings, it is to be understood that one of the two residues is selective, i. H. 



  to the exclusion of the other rest, can react.  So X and Y or  X 'and Y' both simultaneously mean, for example, carboxy; likewise, they cannot both represent carboxy esterified at the same time.  In contrast, z. B. 



  one of the substituents X and Y or  X 'and Y' represent carboxy and the other esterified carboxy because the carboxy group can be selectively reduced to the hydroxymethyl group (cf.  below).  X and Y or  X 'and Y' cannot both represent hydroxymethyl or etherified or esterified hydroxymethyl simultaneously, nor combinations of these groups; however, one of the substituents X and Y or  X 'and Y' are alkyl-etherified hydroxymethyl and the other optionally esterified hydroxymethyl, because alkoxymethyl remains in the case of saponification, halogenation and magnesium or  Organophosphorus reactions in contrast to optionally esterified hydroxymethyl obtained. 



   The under the meaning of X and Y or  X 'and Y' listed, optionally esterified carboxy groups, optionally etherified or esterified hydroxymethyl groups or  acetalized formyl groups are conventional groups.  Esterified carboxyl radicals are, for example, radicals of the formula RIOOC-, in which Rl represents lower alkyl, phenyl or phenyl-lower alkyl.  Esterified / etherified hydroxymethyl radicals are, for example, radicals of the formula R20CH2-, in which R2 is a lower acyl group, e.g. 

  B.  lower alkanoyl, benzoyl or phenyl-lower alkanoyl, or an ether protecting group, e.g. B.  is lower alkyl, or radicals of the formula R30-CHR4-, in which R3 is lower alkyl and R4 is hydrogen or lower alkyl or together with R3 n butylene (2-tetrahydropyranyl), acetalized formyl groups are, for example, radicals of the formula
EMI2. 1
 in which Rs denotes lower alkyl, or both substituents Rs together represent lower alkylene.  In the residues X and Y or  X 'and Y' are any lower alkyl, lower alkoxy, lower alkylene and lower alkanoyl groups, alone or in compositions [such as phenyl-lower alkyl, phenyl-lower alkanoyl, di- (lower-alkoxy) -methyl] straight-chain or branched groups, which preferably contain up to 4 carbon atoms. 

  Examples are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec butyl, t-butyl; Methoxy, ethoxy, n-propoxy, isopropoxy, n butoxy, isobutoxy, sec-butoxy, t-butoxy; Methylene, ethylene, n-propylene, 1-methyl-ethylene, 1,2-dimethyl-ethylene; Acetyl, propionyl, n-butyryl, isobutyryl. 



   X and Y in the meaning carboxy or formyl are preferably esterified or  acetalized.  Preferred groups R in esterified carboxy groups RlOOC- are methyl and ethyl.  Preferred groups Rs in acetalized formyl groups
EMI2. 2nd
 are methyl and ethyl and (both Rs together) ethylene and 1,3-propylene. 



   X and Y with the meaning hydroxymethyl can be unsubstituted or etherified or  be esterified.  If the hydroxymethyl group is substituted, the substituent is preferably methyl, acetyl or benzoyl. 



   The microorganism used according to the invention has an aerobic or facultative aerobic character, ie. H.  it has the ability to grow both under aerobic and under anaerobic conditions (optionally aerobic microorganisms) or even only under aerobic conditions (aerobic microorganism).  By testing any aerobic or optional aerobic yeast, fungi and bacteria for the starting material of formula I used according to the invention, it is easy to find a suitable microorganism which is able to hydrogenate the double bond between CH and methylated C and is therefore used in the process according to the invention can be.  Known microorganisms which can preferably be used are: 1) Eukaryotes 1. 1.  Yeasts of the genus
Candida e.g.  B.  C.  albicans C.  gufflermondii
Kloeckera z.  B.  K.  brevis
Rhodotorula e.g.  B.  R.  rotundata
Saccharomyces e.g.  B.  

  S.  carlsbergensis
S.  cerevisiae 5.  cerium.  ellipsoides
Torulopsis z.  B.  T.  apicola
T.  rotundata 1 2nd  Mushrooms of the genus
Absidia e.g.  B.  A.  rainy
Aspergillus e.g.  B.  A.  clavatus
A.  fumigatus
A.  ochraceus
A.  Niger
Colletotrichum z.  B.  C.  gloeosporioides
Cunninghamella e.g.  B.  C.  blakesleeana curvularia z.  B.  C.  lunata
Geotrichum z.  B.  G.  candidum
Gibberella e.g.  B.  G.  fujikuroi
Gliocladium e.g.  B.  G.  roseum
Mucor z.  B.  M.  circinelloides
M.  corymbifer
M.  griseo-cyanus
M.  hiemalis
M.  parasiticus
M.  spinosus
Penicillium e.g.  B.  P.  notatum
P.  novae-zeelandiae P. virid
Phycomyces e.g.  B.  P.  blakesleeanus
Rhizopus z.  B.  R.  arrhizus
R.  circinans
Trichothecium z.  B.  T.  roseum 2  Prokaryotes 2. 1.  Mycelium-forming bacteria (Actinomycetes) of the genera
Mycobacterium e.g.  B.  M.  butyricum
M.  rhodochrous
Streptomyces e.g.  B. 

  S.  fradiae
S.  rimosus
Proactinomyces e.g.  B.  P.  restrictus
P.  roseus 2 2nd  Gram-positive bacteria of the genus
Bacillus e.g.  B.  B.  subtilis
Micrococcus e.g.  B.  M.  lysodeikticus Propionibacterium e.g.  B.  P.    shermanu
Pediococcus e.g.  B.  P.  cerevisiae
Streptococcus e.g.  B.  S.  faecalis
S.  lactis 2. 3rd  Gram-negative bacteria of the genus
Azotobacter e.g.  B.  A.  indicus
Pseudomonas e.g.  B.  P.  fluorescens
Serratia e.g.  B.  S.  marcescens
Vibrio z.  B.  V.  metschnikovii
Preferred microorganisms in the process according to the invention are, as can be seen from the explanations below, Saccharomyces cerevisiae (press yeast, baker's yeast) and Geotrichum candidum.  Saccharomyces cerevisiae is commercially available as press yeast.  Geotrichum candidum is also a well-known microorganism that is generally accessible from recognized deposits. 

  To ensure the feasibility of the method using Geotrichum candidum, however, a culture of the strain used was carried out at the Centraalbureau voor Schimmelcultures in Baarn, Holland under the number CBS 233. 76 deposited. 



   It goes without saying that each of the microorganisms used according to the invention should be grown before being used in the fermentation according to the invention; cultivation is usually carried out in a manner known per se in an aqueous medium with the aid of the usual nutrients, ie. H.  in the presence of a carbon source such as glucose, fructose.     Sucrose and / or maltose; a nitrogen source such as urea, peptone, yeast extract, meat extract, amino acids and / or ammonium salts; inorganic salts such as magnesium, sodium, potassium, calcium and / or ferrous salts; other growth-promoting substances, such as vitamins and the like. 



  Sometimes it is expedient to also use the cultivation medium in the fermentation according to the invention, although - as explained in more detail below - the composition of the fermentation medium used according to the invention can be considerably simpler. 



   The fermentation according to the invention can be carried out without further additives using only the starting material of the formula I and the microorganism to be used.  However, it is advantageous to add an assimilable carbon source as a microorganism nutrient to the aqueous medium, preferably in an amount of about 10 to 100 g per liter, for example in the form of a sugar such as glucose, fructose, sucrose, maltose and the like, so that the viability and the associated metabolic activity of the microorganism is preserved as long as possible.  More than 100 g of carbon source per liter of nutrient medium does not affect the end result, but does not offer any advantages over the case where 10 to 100 g of carbon source are added. 

  In the case of long fermentation times, it is advantageous to refill the carbon source once or more times in a preferred amount of 10 to 100 g / l.  The addition of a nitrogen source is not necessary; however, an assimilable nitrogen source can optionally be added, preferably in an amount of about 1 to 50 g per liter, for example in the form of urea, peptone, yeast extract, meat extract, amino acids, ammonium salts and the like. 



  The fermentation medium can also include inorganic salts such as magnesium, sodium, potassium, calcium and / or ferrous salts, other growth-promoting substances such as vitamins and the like. , contain. 



   The pH of the fermentation should preferably be within the range from 2 to 10, in particular 3 to 8, a range which can usually be achieved without special additives. 



  If desired, the pH can be adjusted by using buffers, e.g. B.  Phosphate, phthalate or tris buffers [tris (hydroxymethyl) aminomethane] can be regulated.  The temperature can vary widely, e.g. B.  between 10oC and 40C, a temperature of 20 to 35oC is preferred.  In order to obtain optimum yields, it is advantageous for the starting material of the formula I to be present in the fermentation broth in a concentration of 0.1 to 5.0%, in particular 0.5 to 2.5%. 



   After the reaction, reactant can be added again in a preferred concentration between 0.2 and 1.5%.  This process can be repeated until the microorganisms are inactivated.  The substrate can also be added continuously using a pump. 



   The useful fermentation time depends on the microorganisms used.  When the starting material is added once, it usually fluctuates between 4 and 250 hours, in particular between 1 and 2 days.  If the educt is added several times or continuously, the fermentation time can be extended accordingly. 



   The fermentation is preferably carried out aerobically, e.g. B.  with stirring, shaking with air or using a ventilation device.  To combat foam, the usual anti-foaming agents, such as silicone oils, polyalkylene glycol derivatives, soybean oil and the like. , are added.  A microorganism which is in the non-growing (stationary) phase is preferably used.  The choice of a stationary microorganism has the advantage that the fermentation process does not have to be carried out under sterile conditions, provided that a nutrient medium is used that does not allow a substantial multiplication of microorganisms, for example a nutrient medium without a nitrogen source. 



   Depending on the choice of the starting material of formula I and the microorganism, this is subjected to different conversions, i. H.  various products of formula II are obtained.  The method according to the invention can be divided into five subgroups A, B, C, D and E according to these aspects:
A) When using starting materials of the general formula
EMI4. 1
 where XA is optionally esterified carboxy and YA is optionally acetalized formyl or XA is esterified carboxy and YA is optionally etherified or esterified hydroxymethyl, selection of suitable microorganisms gives fermentation products of the formula II in which X 'is optionally esterified carboxy and Y' is optionally etherified or esterified hydroxymethyl represents, free carboxy and hydroxymethyl groups are lactonized with each other to form the group -CO-O-CH2-. 

  The lactonized product is the (S) -dihydro-4-methyl-2- (3H) -furanone of the formula
EMI4. 2nd

The fermentation brings about the saturation of the double bond to form an optically uniform compound and also the reduction of the formyl group or  Hydrolysis and reduction of the acetalized formyl group to hydroxymethyl. 



  Any substituents on carboxy or  Hydroxymethyl are less easily hydrolyzed.  These groups can initially be partially preserved after the double bond is saturated.  Corresponding fermentation products of formula II, which are still substituted on carboxy and / or hydroxymethyl, can be isolated as such from the fermentation broth, or they can by prolonging the fermentation period, for. B. 



  up to 3 to 10 days, are converted into the lactone of the formula IIA by the substituents on carboxy or 



  Hydroxymethyl be cleaved.  The fermentation product undergoes lactone ring closure to form the lactone of formula IIA defined above.  Lactone ring closure can also occur during the purification of a fermentation product of formula II in which X 'is esterified carboxy and Y' is hydroxymethyl (e.g. B.  by distillation, preferably under slightly acidic, e.g. B.  p-toluenesulfonic acid conditions).  The splitting of the substituents on carboxy or  Hydroxymethyl can also be made by alkaline saponification. 



   If an alkyl etherified, for. B.  represents methyl etherified, hydroxymethyl group, this alkyl group is retained after fermentation.  Fermentation products of the formula II are obtained in which X 'is optionally esterified carboxy and Y' is alkoxymethyl. 



   Saccharomyces cerevisiae (pressed yeast, baker's yeast) is preferably used as the microorganism for the fermentation of the starting materials of the formula IA.  According to a preferred embodiment, a starting compound of the formula IA is used in which XA is optionally esterified carboxy and YA is optionally acetalized formyl.  The following microorganisms can be used for this embodiment, for example: 1.  Eukaryotes 1. 1.  Yeast of the genus
Candida e.g.  B.  C.  utilis
Kloeckera z.  B.  K.  brevis
Saccharomyces e.g.  B.  S.  carlsbergensis
S.  cerevisiae
S.  cerium.  ellipsoides
Torulopsis z.  B.  T.  apicola 1 2nd  Mushrooms of the genus
Absidia e.g.  B.  A.  rainy
Aspergillus e.g.  B.  A.  Niger
Colletotrichum z.  B.  C.  gloeosporioides
Cunninghamella e.g.  B.  C.  blakesleeana
Gibberella e.g.  B.  G.  fujikuroi
Gliocladium e.g.  B. 

  G.  roseum
Mucor z.  B.  M.  circinelloides
M.  griseo-cyanus
M.  parasiticus
Penicillium e.g.  B.  P.  novae-zeelandiae
Phycomyces e.g.  B.  P.  blakesleeanus
Rhizopus z.  B.  R.  arrhizus
R.  circinans
Trichothecium z.  B.  T.  roseum 2  Prokaryotes 2. 1.  Mycelium-forming bacteria (Actinomycetes) of the genus
Mycobacterium e.g.  B.  M.  butyricum 2. 3rd  Gram-negative bacteria of the genus
Azotobacter e.g.  B.  A.  indicus
According to a particularly preferred process, ethyl trans-4,4-dimethoxy-3-methylcrotonate is used as the starting compound of the formula I and Saccharomyces cerevisiae is used as the microorganism, ethyl (S) -3-methyl-4-hydroxy-butyrate being obtained primarily as the fermentation product of the formula II becomes.  Acid hydrolysis of this compound in the manner described above provides the (S) -dihydro-4methyl-2 (3H) -furanone. 

 

   B) When using starting materials of the general formula
EMI4. 3rd
 in which one of the radicals XB and YB is carboxy and the other is esterified carboxy, fermentation products of the formula II are obtained when suitable microorganisms are selected, in which one of the radicals X 'and Y' is carboxy and the other is esterified carboxy.  Only the stereospecific saturation of the double bond is thus brought about.  Saccharomyces cerevisiae is preferably used as the microorganism for this reaction.  The starting compound of the formula IB is in particular a compound in which XB is esterified carboxy and YB is carboxy. 



   C) When using starting materials of the general formula
EMI5. 1
 where Xc is esterified carboxy and Yc is optionally acetalized formyl or optionally esterified hydroxymethyl, in addition to the stereospecific saturation of the double bond, an oxidation or  Hydrolysis and oxidation of the optionally acetalized formyl group or the optionally esterified hydroxymethyl group occur.  In this case, fermentation products of formula II are obtained in which X 'is esterified carboxy and Y' is carboxy.  This conversion is preferably triggered by the mushroom Geotrichum candidum. 



   D) When using starting materials of the general formula
EMI5. 2nd
 where XD is optionally esterified hydroxymethyl and YD is optionally acetalized formyl or esterified carboxy, the choice of suitable microorganisms gives fermentation products of the formula II in which X 'is optionally esterified hydroxymethyl and Y' is optionally esterified carboxy, free carboxy and hydroxymethyl groups being formed with one another of the group -CH2-O CO- lactonize.  The lactonized product is the (S) -dihydro-3-methyl-2 (3H) -furanone of the formula
EMI5. 3rd

Thus, on the one hand, a stereospecific saturation of the double bond is brought about, on the other hand existing, optionally acetalized formyl groups are oxidized or  subjected to hydrolysis and oxidation. 



   The optionally present substituents on hydroxymethyl or  As with subgroup A) above, carboxy is less easily hydrolyzable than the substituents on formyl and can initially be partially preserved after the double bond has been saturated.  They can be cleaved by prolonging the fermentation to give the lactone of formula IID. 



   For this embodiment of the method according to the invention, the fungus Geotrichum candidum is preferably used as the microorganism.  According to a preferred embodiment, a starting compound of the formula ID is used, in which XD is optionally esterified hydroxymethyl (preferably free hydroxymethyl) and YE is optionally acetalized formyl (preferably acetalized formyl).  The following microorganisms can be used for this embodiment, for example: 1.  Eukaryotes 1. 1.  Yeasts of the genus
Candida e.g.  B.  C.  albicans
C.  guillermondii
Rhodotorula e.g.  B.  R.  rotundata
Saccharomyces e.g.  B.  S.  cerevisiae
Torulopsis z.  B.  T.  apicola
T.  rotundata 1 2nd  Mushrooms of the genus
Aspergillus e.g.  B.  A.  clavatus
A.    fischeri
A.  flavus
A.  fumigatus
A.  ochraceus Less
Curvularia z.  B.  C.  lunata
Cylindrocarpon z.  B. 

  C.  radicicola
Mucor z.  B.  M.  corymbifer
M.  griseo-cyanus
M.  hiemalis
M.  parasiticus
M.  spinosus penicfflium z.  B.  P.  notatum
P.  virid
Rhizopus z.  B.  R.  arrhizus
R.  circinans
Absidia e.g.  B.  A.  rainy
Geotrichum z.  B.  G.  candidum
Gibberella e.g.  B.  Gibberella fujikuroi
Gliocladium e.g.  B.  G.  roseum
Phycomyces e.g.  B.  P.  blakesleeanus 2  Prokaryotes 2. 1.  Mycelium-forming bacteria (Actinomycetes) of the genera
Mycobacterium e.g.  B.  M.  butyricum
M.  rhodochrous
Streptomyces e.g.  B.  5.  fradiae
S.  rimosus
Proactinomyces e.g.  B.  P.  restrictus
P.  roseus 2 2nd  Gram-positive bacteria of the genus
Bacillus e.g.  B.  B.  subtilis
Micrococcus e.g.  B.  M.  lysodeikticus Propionibacterium e.g.  B.  P.    shermanii
Pediococcus e.g.  B.  P.  cerevisiae
Streptococcus e.g.  B.  S.  faecalis
S.  lactis 2. 3rd 

  Gram-negative bacteria of the genus
Pseudomonas e.g.  B.  P.  fluorencens
Serratia e.g.  B.  S.  marcescens
Vibrio z.  B.  V.  metschnikovü - According to a particularly preferred process, trans-3- (1,3-dioxolan-2-yl) -2-buten-1-ol is used as the starting compound of the formula I and Geotrichum candidum as the microorganism, the (S) being the fermentation product -Dihydro3-methyl-2 (3H) -furanone of the formula IID is obtained. 



   If, in formula ID, XD represents esterified hydroxymethyl and YD esterified carboxy, Geotrichum candidum also provides the end product indicated above, but higher yields are obtained by using Saccharomyces cerevisiae. 



   E) When using starting materials of the general formula
EMI6. 1
 where XE represents alkyl-etherified hydroxymethyl and YE optionally acetalized formyl or optionally esterified hydroxymethyl, fermentation products of the formula II are obtained when suitable microorganisms are selected, where X 'is alkyl-etherified hydroxymethyl and Y' carboxy or optionally esterified hydroxymethyl.  It is characteristic of this educt group that the alkoxymethyl group XE remains unchanged during the fermentation. 



   With Saccharomyces cerevisiae, for example, a reduction or  Hydrolysis and reduction of the optionally acetalized formyl group YE TO hydroxymethyl, while esterified hydroxymethyl groups YE can be partially hydrolyzed and hydroxymethyl groups YE remain unchanged.  With Geotrichum candidum an oxidation or  Hydrolysis and oxidation of the optionally acetalized formyl group and also the optionally esterified hydroxymethyl group YE in carboxy. 



   After the cultivation has ended, the fermentation product is isolated from the fermentation broth in the customary manner.  Extraction with a water-insoluble organic solvent is preferred, for example with an aliphatic or cycloaliphatic, optionally chlorinated hydrocarbon, such as n-hexane, cyclohexane, methylene chloride, chloroform, carbon tetrachloride; an aliphatic ester such as ethyl acetate, n-butyl acetate, amyl acetate; or an aliphatic ether such as diethyl ether or diisopropyl ether.  A preferred solvent is methylene chloride.  To avoid emulsions, the extraction is expediently carried out continuously or with the aid of the multiplicative distribution. 

  According to a preferred isolation method, the fermented broth is first filtered or centrifuged, after which the aqueous phase and the sediment are worked up separately.  The crude product obtained can in the usual manner, for. B.  by fractional distillation.  As already mentioned, open intermediate products of the formula II (with the exception of the alkyl ethers) obtained can be worked up into the corresponding lactone of the formula IIA or  IID pass over. 



   The product of formula II obtained can be converted into optically active vitamin E or K1, for example as follows:
Ester groups on hydroxymethyl or  carboxy can be split off hydrolytically, preferably by basic hydrolysis.  The corresponding lactone is obtained.  In order to obtain optically pure lactone, those esters which lead to the lactone of the formula IIA are preferably used for the hydrolysis. 



   Fermentation products of formula II in which one of X 'and Y' is carboxy and the other is esterified carboxy (sub-group B above) can be selectively reduced chemically, converting the free carboxy group to hydroxymethyl.  This reduction takes place e.g. B.  by treatment with a borohydride / dimethyl sulfide complex, preferably in an ethereal solvent such as tetrahydrofuran, and at a temperature from about -10oC to room temperature.  The carboxy protecting group can be split off in a manner known per se, the lactone of the formula IIA or  IID receives.  Preferably, the selective reduction is carried out on a fermentation product of formula II, wherein X 'is esterified carboxy and Y' is carboxy; the lactone of the formula IIA is obtained. 

 

   The lactones of the formulas IIA and IID can be converted into natural, optically active vitamin E, into optically active vitamin E esters and into optically active vitamin Kl according to the following reaction scheme:
EMI6. 2nd
  
EMI7. 1
  
EMI8. 1
 (Vitamin E or  its ester)
EMI8. 2nd
  
EMI9. 1
 (Vitamin K1)
In the above reaction scheme, R6 represents a carboxyl protecting group, preferably lower alkyl, e.g. B.  Methyl or ethyl; R7 is a carbonyl-free protective group which can be split off by acid hydrolysis, preferably 2-tetrahydropyranyl; Rs is hydrogen or lower alkanoyl, e.g.  B.  Acetyl; Rs lower alkyl, e.g. B.  Methyl or ethyl; Rlo the group
EMI10. 1
   Rll lower alkyl, e.g. B.  Methyl or ethyl; R12 is hydrogen or lower aryl, e.g. B. 

  Acetyl or benzoyl and Hal is a halogen atom, e.g. B.  Chlorine, bromine or iodine. 



   The reactions IIAIII and IIDeVIII take place by treatment with hydrogen halide, preferably hydrogen chloride or hydrogen bromide, in an alcohol corresponding to the introduction of the protective group R6, for. B.  in a lower alkanol such as methanol or ethanol.  Instead of the lactones IIA and IID, the corresponding open fermentation products of the formula II can also be used, in which one of the substituents X1 and Y 'is esterified carboxy and the other hydroxymethyl. 



   The ester group of compounds III is reductively converted to hydroxymethyl to form compounds IV.  The reduction takes place e.g. B.  at -20 to OoC in an inert organic solvent with an organoaluminum compound, e.g. B.  with an ss-branched aluminum di-lower alkyl hydride, such as di-isobutyl aluminum hydride or also with lithium aluminum hydride. 



   By milder reduction of the ester group in compounds VIII, this is converted into the aldehyde group to form compounds IX, for example by treatment with an ss-branched aluminum di-lower alkyl hydride, such as di-isobutyl aluminum hydride at -80 to -400C .    



   The hydroxyl group of compounds IV is protected with the formation of compounds V by a protective group R7 which can be split off by acid hydrolysis.  The introduction of these protective groups takes place e.g. B.  by treatment with the appropriate olefinic compound, e.g. B.  with 3,4-dihydro-2,4-pyran, methyl vinyl ether or 2-methyl propene. 



   The chain extension of the Cs compound of the formula V obtained can be reacted with a 1-halo-3-methylbutane, preferably with 1-bromo-3-methylbutane, or with a corresponding sulfonate, preferably 3-methyl-1butanol-p-toluenesulfonate, with the help of an organomagnesium reaction.  A bromide of the formula V is preferably used.  To carry out the organomagnesium reaction, the compound of formula V with metallic magnesium in an ethereal solvent, for. B.  in tetrahydrofuran, preferably at a temperature between about room temperature and 80 C, wherein magnesium is added. 

  After adding the second coupling component and a catalytic amount of a di (alkali metal) tetrahalo cuprate, preferably dilithium tetrachloro cuprate, the desired Clo compound of the formula VI is obtained.     The ether solvent mentioned is used as the solvent.  The temperature is generally between about -800C and room temperature.  The reaction is preferably initiated at a low temperature (-80 to 0OC) and then at a higher temperature, e.g. B.  Room temperature, completed. 



   In the product of formula VI obtained, the group R7S is now exchanged for halogen, in particular for bromine, chlorine or iodine.  This is preferably done by acid hydrolysis, e.g. B.  with aqueous mineral acid, such as hydrochloric acid, phosphoric acid or sulfuric acid at room temperature, followed by halogenation with the corresponding hydrogen halide, or also by treatment with N-chlorine or N-bromosuccinimide and triphenylphosphine, with phosphorus tribromide or  chloride or phosphorus pentabromide or 



  chloride, with triphenylphosphorodibromide or    -chlond or with methyltriphenoxyphosphoniumj odid.  A bromo derivative of formula VII obtained can be converted into the corresponding iodine derivative by treatment with an alkali metal iodide in a ketonic solvent, e.g. B.  Methyl ethyl ketone or methyl isobutyl ketone, can be converted at elevated temperature. 



   Thereafter, for further chain extension, the compounds V and VII are reacted with one another, with the aid of an organomagnesium reaction, essentially in the same manner as described above for the chain extension V <VI. This gives an optically active Cls compound of the formula XI which is converted into the halide XII in the same way as the conversion VVI just described.



   The chain extension of the aldehyde IX takes place with the aid of an organophosphorus reaction by reaction with a 3-methylbutyl-triarylphosphonium halide, preferably 3-methylbutyl-triphenylphosphonium bromide. The reaction takes place in the presence of a base, e.g. in the presence of a lower alkyl lithium such as n-butyllithium, phenyllithium, or in the presence of an alkali methyl hydride or amide, optionally in an inert organic solvent, e.g. in a lower alkane, such as n-hexane; in a chlorinated hydrocarbon such as methylene chloride; in an ether such as diethyl ether; in an aprotic, polar solvent, such as dimethyl sulfoxide, dimethylformamide or hexamethylphosphoric triamide or in mixtures of these solvents, at a temperature between about -10 ° C. and room temperature.



   For further chain extension, the compounds IX and X are reacted with one another, specifically with the aid of an organophosphorus reaction, essentially in the same manner as indicated above for the chain extension IX <X.



  In this way, the optically active halide XV is obtained.



   The halides XII and XV obtained can be converted into natural, optically active with the aid of the described organophosphorus reaction with (S) -6 hydroxy-2-formyl-2,5, 7,8-tetramethyl-2-chroman or with the corresponding 6-ester Vitamin E or its 6-ester (formula XIV) are implemented. The corresponding unsaturated compounds XIII and XVI with 1 or 3 double bonds in the side chain are obtained first. These double bonds can be saturated by catalytic hydrogenation. A noble metal catalyst which is usually used for hydrogenations, e.g. Platinum dioxide, palladium-carbon, but also Raney nickel or Raney cobalt. The hydrogenation is preferably carried out in a lower alkane carboxylic acid ester, such as ethyl acetate, or in a lower alkanol, such as methanol or ethanol.

  It is carried out in particular under normal pressure and at a temperature between room temperature and about 80OC.



  Any product obtained with a free hydroxy group in the 6-position of the chromium portion can, if desired, be esterified, e.g. with acetic anhydride in pyridine.



   Lactones IIA and IID can also be used to produce natural, optically active vitamin K1. Proceeding from the halides XV and XII obtained in the above manner, one can proceed as follows:
The halides XV are converted into the halides XII by hydrogenation as in the reactions XIIIoXIV and XVIoXIV. A halide XII is by the action of a lower alkyl acetoacetic acid, e.g. B. the methyl or ethyl ester, converted into the ester XVII, which is hydrolyzed and decarboxylated with aqueous alkali to hexahydrofarnesylacetone of the formula XVIII.



   In one variant, the optically active hexahydrofarnesylacetone XVIII is converted into optically active isophytol XX by reaction with an alkali metal acetylide, followed by partial hydrogenation with Lindlar catalyst. Optionally, the hexahydrofarnesylacetone XVIII with a compound of the formula
EMI11.1
 in a lower alkanol solvent and the corresponding lower alkali metal alkoxide, e.g. with triethylphosphonoacetate in ethanolic sodium ethoxide, to give compound XXI, which is converted into optically active phytol XXII by reduction with a complex metal hydride, such as lithium aluminum hydride or diisobutylaluminium hydride.



   The isophytol XX or the phytol XXII can be extracted using a Lewis acid, e.g. Boron trifluoride, in an ethereal solvent, e.g. Dibutyl ether to be condensed with menadiol or its 1 acyl derivative (compound XXIII). After saponification of the 1-acyl derivative XXIV obtained, e.g. with methanolic alkali, the optically active vitamin Kl of formula XXV is obtained by oxidation with air. The proportion of the desired trans product can be increased by separating the cis and trans forms XXI and / or XXIV, for example by chromatography or recrystallization.



   The conversion of optically active hexahydrofarnesylacetone into natural phytol and natural vitamin Kl is also described in J. Chem. Soc. (C), 1966, pages 2144-2176 (in particular 2146, 2151 and 2152) or in Helv. Chim. Acta, 48, 1965, pages 1332-1347 (especially 1333 and 1346).



   Fermentation products of the formula
EMI11.2
 in which X "represents alkyl-etherified hydroxymethyl, eg methoxymethyl, and Y" represents optionally esterified carboxy or optionally esterified hydroxymethyl, the corresponding compounds of formula II ", in which Y" represents hydroxymethyl, can be converted, the optionally esterified carboxy group in analogy to the above reaction III-iIV and the esterified hydroxy group is saponified. The alcohol of formula IIi can be converted to the corresponding tosylate using tosyl chloride.

  This can be done with a 1-halo-3-methylbutane, e.g. 1-Bromo-3-methyl-butane and metallic magnesium are converted into the corresponding Clo compound with the aid of dilithium tetrachlorocuprate under reaction conditions as described for the reaction VVI, after which the alkoxy group against halogen to form the above compound VII is exchanged. By treatment with boron tribromide or chloride in methylene chloride at about 200 to 0OC, alkoxy changes into the hydroxy group, which is then halogenated as described above.



  The compound VII is converted into vitamin E, vitamin E ester or vitamin Kl in the above manner.



   The following depositories are listed in the examples below: ATCC = American Type Culture Collection,
Rockville, Maryland, USA CBS = Centraal-Bureau voor Schimmelcultures,
Baarn - Holland NRRL = Northern Utilization Research and
Development Division of U.S.D.A.,
Peoria, Illinois, USA NCIB = National Collection of Industrial Bacteria,
Aberdeen - Scotland ETH = Swiss Federal Institute of Technology,
Zurich - Switzerland PRL = Prairie Regional Laboratories, Sascatoon, Canada All temperatures are given in degrees Celsius.



   example 1
Transformation of trans-3- (1,3-dioxolan-2-yl) -2-buten-1-ol to (S) -dihydro-3-methyl-2 (3H) -furanone.



   Biocatalyst: Geotrichum candidum.
EMI11.3




   Cultivation of the microorganism
The microorganism is grown in a medium of the following composition:
EMI11.4


 <tb> 20 <SEP> g <SEP> D (+) - glucose <SEP> (monohydrate) <SEP> | <SEP> in <SEP> one <SEP> liters
 <tb> 10 <SEP> g <SEP> Yeast extract <SEP> (Difco) <SEP> deionized
 <tb> 16 <SEP> g <SEP> KH2PO4 <SEP> water
 <tb> <SEP> 2.6 <SEP> g <SEP> Na2HPO4 <SEP> ' <SEP> l <SEP> (pH <SEP> 6)
 <tb>
This medium is sterilized in an autoclave at 1350 for 30 minutes. The microorganism is inoculated from a slant agar culture in 500 ml of culture media using the usual microbiological working technique and incubated in a sterile Erlenmeyer flask with sterile plugs for 48 hours on a rotary shaker at 30 °.

  This approach is then transferred to a sterile laboratory fermenter containing 20 liters of nutrient medium of the above composition. To combat foam, 10 ml of polypropylene glycol monobutyl ether are added. The fermenter is operated for 24 hours, at a thermostatically controlled temperature of 300, with a stirring frequency of 900 rpm and an air flow rate of 600 l / h. The biomass is then filtered off. 1 kg of biomass is obtained from such a batch. The biomass is kept in the refrigerator until it is used in the transformation experiment.



   Transformation of trans-3- (1,3-dioxolane
2-yl) -2-buten-1-ol (a)
18 l of deionized water and 200 g of sucrose are introduced into a clean, but not sterilized, laboratory fermenter (total volume 311). 2 kg of biomass (Geotrichum candidum CBS 233.76) are suspended in this sugar solution. 350 g of substrate (a) are then added. This batch is aerated for 24 h at a thermostatically controlled temperature of 30 with stirring (stirrer speed 900 rpm) with an air flow rate of 600 l / h. After 2, 4, 6, 8 and 24 h, 10 ml samples are taken, extracted twice with methylene chloride, dried over Na2SO4 and concentrated under reduced pressure.

  The residue is taken up in dioxane and analyzed by gas chromatography (glass column with 12% Carbowax as adsorbent, start temperature 100, temperature rise 4 '/ min, end temperature 220, carrier gas: N2). The percentage transformation to the optically active fermentation product (S) -dihydro-3-methyl2 (3H) -furanone (IID) is shown in Table 1.



   Table fermentation time transformation to lactone IID
2h 11.6% 4h 23.9% 6h 35.7% 8h 44.0% 24h 61.1%
The fermentation is stopped after 24 hours and the desired fermentation product is isolated as follows:
The broth is filtered. The water phase and sediment are processed separately. The water phase is stirred twice with 40 1 methylene chloride, the sediment is shaken twice with 5 1 methylene chloride. The separated solvent phase is dewatered over Na2SO4 and concentrated under reduced pressure. The result is 248 g of crude extract.

  This crude extract is distilled at 81-840 / 14-15 Torr (bath temperature 105-115). 107.8 g of product are obtained, which according to the gas chromatogram consists of 95% of (S) -dihydro-3methyl-2 (3H) -furanone and has an optical rotation of = -20.7 (2% in ethanol). Only one enantiomer (S configuration) is visible in the NMR spectrum of the product, recorded with the addition of chiral shift reagents.



   Example 2
Transformation of different educts to (S)
Dihydro-3-methyl-2 (3H) furanone. Biocatalyst:
Geotrichum candidum CBS 233.76
The microorganism is grown in the same manner as described in Example 1.



   The following substrates are used in the transformation experiments:
EMI12.1

EMI12.2

The transformation experiments are set up as follows: 5 g of biomass (Geotrichum candidum CBS 233.76) are suspended in 45 ml of sterile transformation medium which has the following composition: D (+) glucose (monohydrate) 50 g of KH2PO4 8 g of Na2HPO4 1.3 g
EMI12.3
 in 1 1 deionized water (pH 6)
Now the educt in a concentration of 1 resp. 2 g / l (see table) were added and the mixture was incubated for 7 days on a rotary shaker (260 rpm) in a thermostated room at 30O. After 1 and 7 days, a 10 ml sample is taken and extracted twice with methylene chloride.

  The solvent phase is separated off, dried over Na2SO4 and concentrated under reduced pressure.



  The residue is taken up in so much dioxane that, based on the starting material used, a 1% solution is formed. This is then analyzed by gas chromatography. The results are shown in the table below.



  table
EMI13.1


 <tb> <SEP> Ä
 <tb> <SEP> educt <SEP> educt contact <SEP> in <SEP>% <SEP> after <SEP> gas chromatogram
 <tb> <SEP> centering <SEP> (GC)
 <tb> <SEP> in <SEP> gil <SEP> fermentation <SEP> Fermenta
 <tb> <SEP> time <SEP> time
 <tb> <SEP> 1 day <SEP> 7 <SEP> days
 <tb> <SEP> 1 <SEP> 1 <SEP> 15 <SEP> 33
 <tb> <SEP> HO
 <tb> <SEP> Ho # · Oo <SEP> 2 <SEP> 40 <SEP> 75
 <tb> <SEP> ·
 <tb> <SEP> HO <SEP> 2 <SEP> 32 <SEP> 74
 <tb> <SEP> 0
 <tb> <SEP> 2 <SEP> 2 <SEP> 89 <SEP> 95
 <tb> <SEP> HO <SEP> O
 <tb> <SEP> o
 <tb> 2 <SEP> 0> <SEP> 74 <SEP> 94
 <tb> Ho
 <tb> <SEP> 1 <SEP> 20 <SEP> 79
 <tb> <SEP> 1 <SEP> 24 <SEP> 45
 <tb> <SEP> o
 <tb> <SEP> 2 <SEP> 79 <SEP> 88
 <tb> 4ow <SEP>) <SEP> 2 <SEP> 79 <SEP> 88
 <tb> <SEP> OIo
 <tb> <SEP> 4o <SEP> H <SEP> 2 <SEP> 76 <SEP> 89
 <tb> tow <SEP> 1 <SEP> 17 <SEP> 55
 <tb> <SEP> O
 <tb>
Example 3 Transformation of

   Ethyl trans-4,4-dimethoxy
3-methyl crotonate. Biocatalyst: press yeast
EMI13.2
  
A clean, but not sterilized fermenter with a total volume of 311 is charged with the following ingredients: - Deionized water 9.91 - Pressed yeast 1.1 kg - Sugar 0.55 kg - Ethyl 4,4-dintethoxy-3-methylcrotonate (b) 133 g - polypropylene glycol monobutyl ether 10 ml
The fermentation is carried out under the following conditions: temperature 30 (thermostatically controlled) stirring frequency 1000 rpm air flow rate 600 I pH 3.4-3.8 (no pH control) fermentation time 56 h
After 16, 24, 40, 48 and 56 h, 10 ml samples are taken and extracted twice with methylene chloride.

  The solvent phase is separated off, dried over Na2SO4 and concentrated under reduced pressure. The residue is taken up in so much dioxane that, based on the starting material used, a 1% solution is formed and then analyzed by gas chromatography.

  The results are summarized in the table below: Composition of the extracts in%
EMI14.1


 <tb> Fer <SEP> o <SEP>> <SEP> AbOH <SEP> A <SEP> 4
 <tb> ment, - <SEP> co-.G-l <SEP> 1;
 <tb> time <SEP> 0- <SEP> 0 <SEP> 0
 <tb> 16 <SEP> h <SEP> 23.8 <SEP> 40.6 <SEP> 34.3 <SEP> 0.3
 <tb> 24h <SEP> 11.9 <SEP> 43.4 <SEP> 42.7 <SEP> 0.3
 <tb> 40 <SEP> h <SEP> 3.7 <SEP> 43.3 <SEP> 49.1 <SEP> 0.3
 <tb> 48h <SEP> 1.8 <SEP> 44.4 <SEP> 50.1 <SEP> 0.7
 <tb> 56h <SEP> 0.5 <SEP> 46.9 <SEP> 49.2 <SEP> 1.0
 <tb>
The fermentation is stopped after 56 h. The desired fermentation product is isolated as follows:
The broth is saturated with NaCl and extracted continuously with diethyl ether for 4 days. The solvent is separated off, dried over Na2SO4 and removed again under reduced pressure.

  122 g of crude extract are obtained, the major part of which contains (S) -3-methyl-4-hydroxy-butyric acid ethyl ester. This substance can be purified chromatographically (over silica gel which has been deactivated with 0.5 SO saturated ammonia).



   Example 4
Hydrolysis of the (S) -3-methyl-4-hydroxy-butyric acid ethyl ester and purification of the resulting (S)
Dihydro-4-methyl-2 (3H) furanons. (IIA)
The crude extract obtained according to Example 3 is mixed with 100 mg of p-toluenesulfonic acid and distilled in a nitrogen atmosphere under reduced pressure. The (S) -dihydro-4-methyl-2 (3H) -fura-non formed during the distillation distilled at 86 to 88 C / 14 Torr (bath temperature 125 to 140 '). 26.2 g of product are obtained which, according to GC, contains 93% (S) dihydro-4-methyl-2 (3H) -furanone and has an optical rotation of -210 (4% in methanol).

  The optical purity of the (S) -dihydro-4-methyl-2 (3H) -furanone can be after converting the lactone into the corresponding bromester
EMI14.2
 using the NMR spectrum, recorded with the addition of a chiral shift reagent [Eu (HFC) 3].



   The yield of isolated, optically pure (S) -dihydro-4methyl-2 (3H) -furanone is 34.4% of the theoretically possible amount of product.



   Example 5
Transformation of ethyl-trans-4,4-dimethoxy-3 methyl-crotonate with continuous substrate supply.



   Biocatalyst: press yeast.



   Two transformation experiments A and B are carried out in a clean, but not sterilized, small fermenter (working volume 5 l) with continuous feeding of the substrate (ethyl-trans-4,4-dimethoxy-3-methyl-crotonate).



  In both cases, the fermenter is charged with the following substances.



  Deionized water (non sterile) 4.51 crystalline sugar 250 g pressed yeast 500 g
Both transformations are carried out under the following conditions: temperature 30'C stirring frequency 1000 rpm air flow rate 600 I pH 3.0-4.3 (no control)
The substrate is continuously pumped in at a delivery rate of approx. 7.5 ml / h. At A the substrate is fed in for 10 h and a final concentration of 16 g / l is reached.



  At B, substrate is pumped in over 22 h, so that the final concentration is 33 g / l.



   In Experiment B, 100 g of sugar are also added after a fermentation time of 17 h.



   To combat foam, 10 ml of polypropylene glycol monobutyl ether are added.



   The course of fermentation A and B is followed by gas chromatographic analyzes according to Example 3.



   Reaction A is largely complete after 20 h, reaction B after 46 h. After this fermentation time, the GC analysis shows the following percentage composition of the extracts:
EMI15.1


 <tb> <SEP> Expen- <SEP> Fermen- <SEP> substrate: <SEP> composition <SEP> the <SEP> extracts <SEP> in <SEP>% <SEP> after <SEP> GC
 <tb> <SEP> ment <SEP> station <SEP> end con <SEP> o <SEP> o <SEP> 0 <SEP> 5
 <tb> <SEP> time <SEP> centering <SEP> two <SEP> Ao) 4 & ( <SEP> Ao) W <SEP> '
 <tb> A <SEP> 20 <SEP> h <SEP> 16 <SEP> g / l <SEP> 5.6 <SEP> 0.4 <SEP> 37.3 <SEP> 53.4 <SEP> 1.6
 <tb> B <SEP> 46 <SEP> h <SEP> 33 <SEP> g / l <SEP> 17.5 <SEP> 6.8 <SEP> 30.0 <SEP> 40.2 <SEP> 2.0
 <tb>
The isolation of the saturated hydroxy ester and the conversion into the (S) -dihydro-4-methyS2 (3H) -furanone are carried out according to Examples 3 and 4.

  18.2 g and 23.0 g of optically pure (S) -dihydro-4-methyl-2 (3H) -furanone were isolated from batch A in this way.



   Example 6
Transformation of various educts to optically active intermediates, which in (S) -Dihydro
4-methyl-2 (3H) furanone can be converted.



   Biocatalyst: press yeast
The following substrates are used in the transformation experiments:
EMI15.2

EMI 15.3

EMI 15.4

EMI15.5

The transformation experiments are carried out in the same way as described in Example 2. However, 5 g of pressed yeast are used as the biocatalyst.

  The results are shown in the following table:
EMI15.6


 <tb> <SEP> educt <SEP> educt ul'in <SEP>%
 <tb> <SEP> concentrated <SEP>,
 <tb> <SEP> tration
 <tb> <SEP> g / l <SEP> Id <SEP> 3d <SEP> 7d
 <tb> <SEP> o
 <tb> <SEP> (" <SEP> 1 <SEP> 14 <SEP> 26 <SEP> 36
 <tb> <SEP> o
 <tb> <SEP> 1 <SEP> 1 <SEP> 69 <SEP> 75 <SEP> 63
 <tb> <SEP>> <SEP>) < <SEP> 2 <SEP> 61 <SEP> 66 <SEP> 58
 <tb> <SEP> o
 <tb> <SEP> 2 <SEP> partly < <SEP> 1 <SEP> 2 <SEP> - <SEP> 12 <SEP> 10
 <tb> <SEP> o
 <tb> <SEP> - <SEP> 2 <SEP> - <SEP> 5 <SEP> 5
 <tb> <SEP> o
 <tb> 1 <SEP> 1 <SEP> 5 <SEP> 22 <SEP> 45
 <tb> <SEP> o
 <tb> <SEP> 2 <SEP> Y <SEP> 2 <SEP> 4 <SEP> 17 <SEP> 34
 <tb>
Example 7
Transformation of trans-3- (1,3-dioxolan-2-yl) -2-buten-1-ol and ethyl-trans-4,4-dimethoxy-3 methylcrotonate by various microorganisms
At least 70 randomly selected microorganisms are tested for their ability to

   to convey the following transformation reactions. tested:
EMI16.1

The microorganisms are selected from the following groups:
1. Eukaryotes
1.1. Genus yeast: Candida
Kloeckera
Rhodotorula
Saccharomyces
Torulopsis 1.2. Mushrooms of the genus: Absidia
Aspergillus
Colletotrichum
Cunninghamella
Curvularia
Cylindrocarpon
Endomyces
Fusarium
Gibberella
Gliocladium
Hypomyces
Mucor
Penicillium
Phycomyces
Rhizopus
Trichothecium 2. Prokaryotes 2.1. Actinomycetes of the genera: Actinomyces
Mycobacterium
Nocardia
Proactinomyces
Streptomyces 2.2. Gram-positive bacteria of the
Genera: Arthrobacter
Bacillus
Micrococcus
Pediococcus
Propionibacterium
Sarcina
Streptococcus 2.3.

  Gram-negative bacteria of the
Genera Azotobacter
Klebsiella
Pseudomonas
Serratia
Vibrio
The microorganisms are inoculated into 50 ml of a complex growth medium using the usual microbiological working technique and incubated on a shaker at 30 ° C. for 48 to 72 hours. The medium is composed as follows: KH2PO4 3.7 g Na2HPO4 7.0 g Yeast extract (Difco) 10 g D (+) - glucose (monohydrate) 20 g
EMI16.2
 in a liter of deionized water
After 48 to 72 h, 0.5 g D (+) - glucose (monohydrate) (10 g / l) and 50 mg trans 3- (1, 3-dioxolan-2-yl) - are added to each 50 ml batch. 2-buten-1-ol resp. Trans-ethyl-4,4-dimethoxy-3-methylcrotonate (1 g / l) was added.

  The incubation continues for a week under the same conditions.



  After one and after 7 days, 10 ml of the cell suspension of all batches are extracted twice with methylene chloride.



  The organic phase is dried over Na2SO4 and concentrated at 40O under reduced pressure. The residue is taken up in 1 ml of dioxane and analyzed by gas chromatography. The results are summarized in the table below. In this table mean - no turnover for the desired product + only traces of the desired product are detected (turnover <5%) + the crude extract contains at least 5% of the desired
Product.



  YEARS
EMI 16.3


 <tb> microorganism <SEP> transforma- <SEP> transformation reaction <SEP> b)
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> K <SEP> FoO <SEP> oX
 <tb> <SEP> 1 day <SEP> 7 days <SEP> 1 day <SEP> 7 days
 <tb> <SEP> Candida <SEP> albicans <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Candida <SEP> guillenmondiii <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Candida <SEP> utilis <SEP> 621 <SEP> CBS <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Kloeckera <SEP> brevis, <SEP> tribe <SEP> 1 <SEP> - <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Kloeckera <SEP> brevis,

    <SEP> tribe <SEP> 2 <SEP> + <SEP> - <SEP> + <SEP> +
 <tb> <SEP> Rhodotorula <SEP> rotundata <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Saccharomyces <SEP> carlsbergensis <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> YEAST (continued)
EMI17.1


 <tb> <SEP> microorganism <SEP> transforma- <SEP> transformation reaction <SEP> h
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> < <SEP> Foffi <SEP> oO
 <tb> <SEP> 1 day <SEP> days <SEP> 1 day <SEP> 7 days
 <tb> Saccharomyces <SEP> cerevisiae, <SEP> tribe <SEP> 1 <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Saccharomyces <SEP> cerevisiae, <SEP> tribe <SEP> 2 <SEP> f <SEP> ¯ <SEP> + <SEP> +
 <tb> Saccharomyces <SEP> cerevisiae, <SEP> tribe <SEP> 3 <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Saccharomyces <SEP> cerevisiae,

    <SEP> ellipsoides <SEP> - <SEP> 9896 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Torulopsis <SEP> apicola <SEP> PRLNo.123-64 <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Torulopsis <SEP> rotundata <SEP> NRRL <SEP> 1402 <SEP> + <SEP> + <SEP> - <SEP> +
 <tb> MUSHROOMS
EMI17.2


 <tb> microorganism <SEP> transforma- <SEP> transformation reaction <SEP> b)
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> X <SEP> Ao4 + oX
 <tb> <SEP> 1 day <SEP> days <SEP> 1 day <SEP> 7 days
 <tb> Absidia <SEP> regneri <SEP> Lendner <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Aspergillus <SEP> clavatus <SEP> + <SEP> + <SEP> i <SEP> +
 <tb> Aspergillus <SEP> fischeri <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Aspergillus <SEP> flavus <SEP> + <SEP> f.
 <tb>



  Aspergillus <SEP> fumigatus <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Aspergillus <SEP> niger <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Aspergillus <SEP> ochraceus <SEP> 12337 <SEP> ATCC <SEP> + <SEP> + <SEP> - <SEP> +
 <tb> Aspergillus <SEP> wentii <SEP> Wehmer <SEP> + <SEP> - <SEP> + <SEP> +
 <tb> Colletotrichum <SEP> gloeosporioides <SEP> + <SEP> - <SEP> + <SEP> +
 <tb> Cunninghamella <SEP> blakesleeana <SEP> + <SEP> i <SEP> + <SEP> +
 <tb> Curvularia <SEP> lunata <SEP> NRRL <SEP> 2380 <SEP> + <SEP> + <SEP> i <SEP> +
 <tb> Cyhndrocarpon <SEP> radicicola <SEP> 11011 <SEP> ATCC <SEP> + <SEP> - <SEP> + <SEP> +
 <tb> Endomyces <SEP> fibular <SEP> 2521 <SEP> CBS <SEP> i <SEP> + <SEP> i <SEP> +
 <tb> Fusarium <SEP> culmorum <SEP> + <SEP> +
 <tb> Fusarium <SEP> solani <SEP> 12823 <SEP> ATCC <SEP> + <SEP> +
 <tb> Gibberella <SEP> fujikuroi <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> gliocladium <SEP> roseum <SEP> +

    <SEP> + <SEP> + <SEP> +
 <tb> Hypomyces <SEP> rosellus <SEP> + <SEP> +
 <tb> Mucor <SEP> circinelloides <SEP> + <SEP> +
 <tb> Mucor <SEP> corymbifer <SEP> + <SEP> + <SEP> ¯ <SEP> +
 <tb> Mucor <SEP> griseo-cyanus <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Mucor <SEP> hiemalis <SEP> + <SEP> +
 <tb> Mucor <SEP> parasiticus <SEP> 6476 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Mucor <SEP> spinosus <SEP> 2604 <SEP> ETH <SEP> + <SEP> +
 <tb> Penicillium <SEP> griseofulvum <SEP> 11 <SEP> 885 <SEP> ATCC <SEP> + <SEP> +
 <tb> Penicillium <SEP> notatum <SEP> 832 <SEP> CBS <SEP> + <SEP> + <SEP> i
 <tb> Penicillium <SEP> novae-zeelandiae <SEP> 10473 <SEP> ATCC <SEP> + <SEP> - <SEP> + <SEP> +
 <tb> Penicillium <SEP> virid <SEP> 2603 <SEP> ETH <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Phycomyces <SEP> blakesleeanus <SEP> + <SEP> + <SEP> i <SEP> +
 <tb> Rhizopus <SEP> arrhizus <SEP> 11 <SEP> 145 <SEP> ATCC <SEP> +

    <SEP> + <SEP> + <SEP> +
 <tb> Rhizopus <SEP> circinans, <SEP> tribe <SEP> 1 <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> Rhizopus <SEP> circinans, <SEP> tribe <SEP> 2 <SEP> - <SEP> i <SEP> + <SEP> +
 <tb> Trichothecium <SEP> roseum <SEP> 8685 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> ACTINOMYCETEN
EMI18.1


 <tb> microorganism <SEP> transforma- <SEP> transformation reaction <SEP> b)
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> 5 <SEP> CH3
 <tb> <SEP> 1 day <SEP> 7 <SEP> days <SEP> 1 day <SEP> 7 <SEP> days
 <tb> Actinomyces <SEP> cellulosae <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> Mycobacterium <SEP> butyricum <SEP> i <SEP> + <SEP> i <SEP> +
 <tb> Mycobacterium <SEP> phlei <SEP> f <SEP> - <SEP> + <SEP> +
 <tb> Mycobacterium <SEP> rhodochrous <SEP> 4277 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP>
 <tb> <SEP> Nocardia <SEP> asteroides <SEP> 27 <SEP> 042 <SEP> ETH <SEP> - <SEP> - <SEP> +

    <SEP> +
 <tb> <SEP> Nocardia <SEP> brasiliensis <SEP> 27 <SEP> 048 <SEP> ETH <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> <SEP> Nocardiaopaca <SEP> 33161 <SEP> CBS <SEP> + <SEP> + <SEP> + <SEP>
 <tb> <SEP> Proactinomyces <SEP> restrictus <SEP> 15 <SEP> 745 <SEP> CBS <SEP> + <SEP> + <SEP> + <SEP>
 <tb> <SEP> Proactinomyces <SEP> roseus <SEP> + - <SEP> + <SEP> + <SEP>
 <tb> <SEP> Streptomyces <SEP> albus <SEP> 6860 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP>
 <tb> <SEP> Streptomyces <SEP> fradiae <SEP> 10 <SEP> 745 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Streptomyces <SEP> lavendulae <SEP> 11 <SEP> 924 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP>
 <tb> <SEP> Streptomyces <SEP> nmosus <SEP> 10970 <SEP> ATCC <SEP> i <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Streptomyces <SEP> venezuelae <SEP> 10 <SEP> 595 <SEP> ATCC <SEP> + <SEP> - <SEP> + <SEP> BACTERIA gram positive
EMI18.2

   


 <tb> microorganism <SEP> transforma- <SEP> transformation reaction <SEP> b)
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> acs
 <tb> <SEP> iTag <SEP> 7 days <SEP> 1 day <SEP> 7 days
 <tb> <SEP> 1 <SEP> day <SEP> 7 <SEP> days <SEP> 1 <SEP> day <SEP> 7 <SEP> days
 <tb> Arthrobacter <SEP> simplex <SEP> 6946 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> Bacillus <SEP> megatherium <SEP> - <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Bacillus <SEP> sphaericus <SEP> 12300 <SEP> ATCC <SEP> - <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Bacillus <SEP> subtilis <SEP> 6633 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Micrococcus <SEP> lysodeikticus <SEP> 4638 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Pediococcus <SEP> cerevisiae <SEP> 8042 <SEP> ATCC <SEP> i <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Propionibacterium <SEP> shermanii <SEP> + <SEP> + <SEP> - <SEP> +
 <tb> <SEP> Sarcina <SEP>

   lutea <SEP> 8340 <SEP> ATCC <SEP> - <SEP> ¯ <SEP> + <SEP> +
 <tb> <SEP> Streptococcus <SEP> faecalis <SEP> 9790 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> <SEP> Streptococcus <SEP> lactis <SEP> + <SEP> + <SEP> + <SEP> +
 <tb> BACTERIA gram negative
EMI18.3


 <tb> <SEP> transforma- <SEP> transformation reaction <SEP> b)
 <tb> <SEP> reaction <SEP> a)
 <tb> <SEP> < <SEP>) 9NotH
 <tb> <SEP> 1 day <SEP> 7 days <SEP> 1 day <SEP> 7 <SEP> days
 <tb> Azotobacter <SEP> indicus <SEP> 9540 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> Klebsiella <SEP> pneumoniae <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> Pseudomonas <SEP> fluorescens <SEP> 13430 <SEP> ATCC <SEP> + <SEP> + <SEP> + <SEP>
 <tb> Pseudomonas <SEP> testosteroni <SEP> 11 <SEP> 996 <SEP> ATCC <SEP> - <SEP> - <SEP> + <SEP> +
 <tb> Serratia <SEP> marcescens <SEP> i <SEP> + <SEP> + <SEP> +
 <tb> Vibrio <SEP> metschnikovii <SEP>

   7708 <SEP> ATCC <SEP> + <SEP> + <SEP> f <SEP> +
 <tb> The results can be summarized as follows:
EMI19.1


 <tb> transformation reaction <SEP> number <SEP> the <SEP> number <SEP> the <SEP> tribes
 <tb> <SEP> checked <SEP> - <SEP> + <SEP> +
 <tb> <SEP> tribes <SEP> no <SEP> required <SEP> less <SEP> at least
 <tb> <SEP> tes <SEP> product <SEP> as <SEP> 5 <SEP>% <SEP> 5 <SEP>% <SEP> product
 <tb> <SEP> proven <SEP> product
 <tb> <SEP> HO + oz <SEP> too <SEP> 11 <SEP> (15.5%) <SEP> 15 <SEP> (21.1%) <SEP> 45 <SEP> (63.4 o)
 <tb> <SEP>: <SEP> W <SEP> 74 <SEP> 0 <SEP> 48 (64.9%) <SEP> 26 (35.1%)
 <tb> <SEP> 0 /
 <tb> <SEP> 0 <SEP> 5,
 <tb>
Example 8 Preparation of the optically active half ester 2-methyl-3-ethoxycarbonyl-propionic acid
EMI19.2

Geotrichum candidum CBS 233.76 is grown as in Example 1.

  The transformation experiments are carried out as in Example 2. The sales of saturated half-ester (according to GC) are tabulated below for fermentation times of 1 3 and 7 days:
EMI20.1


 <tb> educt <SEP> educt contact <SEP> micro <SEP> in <SEP>%
 <tb> <SEP> centering <SEP> organis
 <tb> <SEP> in <SEP> g / l <SEP> mus <SEP> 1 <SEP> day <SEP> 3 <SEP> days <SEP> 7 <SEP> days
 <tb> <SEP> K <SEP> 1 <SEP> press <SEP> 70 <SEP> 92 <SEP> 76
 <tb> <SEP> yeast
 <tb> <SEP> cn
 <tb> <SEP> D <SEP> 1 <SEP> 72 <SEP> 82 <SEP> 78
 <tb> <SEP> V1
 <tb> <SEP> 1 <SEP>) < <SEP> 1 <SEP> 14 <SEP> 38
 <tb> <SEP> 0
 <tb>> <SEP> + <SEP> 2 <SEP> U <SEP> 7 <SEP> 66 <SEP> 84
 <tb> <SEP> ii
 <tb> <SEP> 1 <SEP> like this <SEP> 75 <SEP> 75
 <tb> <SEP> o <SEP> O
 <tb> <SEP> o <SEP> C3
 <tb> <SEP> 2 <SEP> 2 <SEP> 41 <SEP> 77 <SEP> 79
 <tb>
Example 9 Conversion of (S) -2-methyl-3-ethoxycarbonyl

   propionic acid in (S) -dihydro-4-methyl-2 (3H) -furanone
819 mg of the optically active half-ester obtained according to Example 7 are dissolved in 6 ml of absolute tetrahydrofuran in an argon atmosphere and cooled to 0 to -10 ° C.



  After adding 3 ml of trimethyl borate and 10.3 ml of BH3-S (CH3) 2 in tetrahydrofuran (dropwise), the reaction mixture is stirred at -10 to 0O for 50 minutes in an argon atmosphere. After carefully adding 20 ml of methanol, the mixture is evaporated at 40O under reduced pressure. The residue is dissolved in dichloromethane and shaken successively with 30 ml of saturated aqueous sodium bicarbonate solution and twice with 20 ml of aqueous sodium chloride solution. The aqueous phase is extracted three times with dichloromethane. The combined organic phases are dried over magnesium sulfate, filtered and evaporated under reduced pressure. 586 mg of ethyl (S) -3 methyl-4-hydroxybutyrate are obtained as a colorless oil.



   The ethyl (S) -3-methyl-4-hydroxybutyrate obtained is heated with a trace of p-toluenesulfonic acid in methanol for 3 hours under reflux conditions. (S) -Dihydro4-methyl-2 (3H) -furanone is obtained, which boils at about 500 / 0.1 torr.

 

   Example 10
Transformation of trans-1,1, 4-trimethoxy-2-methyl-2-butene, trans-4-methoxy-2-methyl-2-buten-1-ol, or 1-ol acetate, trans-2- (3-methoxy-1-methylpropenyl) - 1,3-dioxolane or trans-4-methoxy-2-methyl-crotonaldehyde by pressing yeast or Geotrichum candidum
Compressed yeast or Geotrichum candidum CBS 233.76 (cultivated as in Example 1) are used as microorganisms. 0.2% of the following are used as substrates
Educts a, b, c, d and e used. The transformation experiments are carried out as in Example 2.

  Depending on the choice of the microorganism, the following main products are produced:
EMI20.2
  
The sales of these products (according to GC = gas chromatogram) are tabulated below for fermentation times of 1, 3 and 7 days:
EMI21.1


 <tb> <SEP> CH3 <SEP> CH3
 <tb> educt <SEP> micro <SEP> Fermenta- \ Ot <SEP> \ <SEP> OtOH
 <tb> <SEP> organism <SEP> time <SEP> 0 <SEP> H
 <tb> <SEP> in <SEP> days <SEP> in <SEP>% <SEP> It.

   <SEP> GC <SEP> in <SEP>% <SEP> lot <SEP> GC
 <tb> <SEP> press yeast <SEP> 1 <SEP> 17 <SEP> 0
 <tb> <SEP> 3 <SEP> 36 <SEP> 7
 <tb> <SEP> OH <SEP> 7 <SEP> 26 <SEP> 32
 <tb> <SEP> 0 <SEP> Geotrichum <SEP> 1 <SEP> 1 <SEP> 42
 <tb> <SEP> candidum <SEP> 3 <SEP> 0 <SEP> 51
 <tb> <SEP> CBS <SEP> 233.76 <SEP> 7 <SEP> 0 <SEP> 64
 <tb> <SEP> press yeast <SEP> 1 <SEP> 13 <SEP> 0
 <tb> <SEP> 3 <SEP> 32 <SEP> 5
 <tb> 7 <SEP> < <SEP> 7 <SEP> 24 <SEP> 25
 <tb> <SEP> 0
 <tb> <SEP> <SEP> Geotrichum <SEP> 1 <SEP> 5 <SEP> 34
 <tb> <SEP> candidum <SEP> 3 <SEP> 1 <SEP> 39
 <tb> <SEP> CBS <SEP> 233.76 <SEP> 7 <SEP> 0 <SEP> 45
 <tb> <SEP> press yeast <SEP> 1 <SEP> 32 <SEP> 0
 <tb> <SEP> 3 <SEP> 54 <SEP> 2
 <tb> <SEP> 0¯ <SEP> 7 <SEP> 7 <SEP> 49 <SEP> 27
 <tb> <SEP> 0o <SEP> Geotrichum <SEP> 1 <SEP> 0 <SEP> 86
 <tb> <SEP> candidum <SEP> 3 <SEP> 0 <SEP> 91
 <tb> <SEP> CBS <SEP> 233.76 <SEP> 7 <SEP> 0 <SEP>

   91
 <tb> <SEP> press yeast <SEP> 1 <SEP> 34 <SEP> 0
 <tb> <SEP> 3 <SEP> 48 <SEP> 0
 <tb> <SEP> 7 <SEP> 7 <SEP> 7 <SEP> 38 <SEP> 36
 <tb> <SEP> Geotrichum <SEP> 1 <SEP> 20 <SEP> 66
 <tb> <SEP> candidum <SEP> 3 <SEP> 0 <SEP> 94
 <tb> <SEP> CBS233.76 <SEP> 7 <SEP> 0 <SEP> 97
 <tb> <SEP> press yeast <SEP> 1 <SEP> 21 <SEP> 0
 <tb> <SEP> 3 <SEP> 43 <SEP> 0
 <tb> <SEP> 7 <SEP> H <SEP> 7 <SEP> 31 <SEP> 39
 <tb> <SEP> Geotrichum <SEP> 1 <SEP> 37 <SEP> 0
 <tb> <SEP> candidum <SEP> 3 <SEP> 1 <SEP> 71
 <tb> <SEP> CBS <SEP> 233.76 <SEP> 7 <SEP> 0 <SEP> 82
 <tb>
Example 11
To 110 ml of a freshly prepared ethanolic bromine hydrogen solution (about 8n) with stirring at 0 "11.0 g (0.11 mol) of (S) -dihydro-4-methyl-2 (3H) -furanone within 10 minutes. The reaction mixture is then stirred for a further two hours at room temperature.

  For working up, the reaction mixture is poured onto ice, diluted to about 1 liter with water and extracted twice with chloroform. The combined organic phases are washed first with water, then with saturated sodium bicarbonate solution until neutral and dried over sodium sulfate. After removal of the chloroform on a rotary evaporator, the colorless ethyl (S) -4-bromo-3-methylbutyrate distilled in a water jet vacuum at 90 to 920. Yield: 18.5 g (80%); [(tJ025 = -2.30 (c = 4.0, CHCl3).



   204 ml (0.204 mol) of a 1 m solution of diisobutylaluminum hydride are added dropwise in an argon atmosphere at 0OC with 17.8 g (0.085 mol) of ethyl- (S) -4-bromo-3-methylbutyrate with stirring within 15 minutes. The excess of the reducing agent is decomposed by dropwise addition of methanol at 0O. The reaction mixture is then poured onto ice and acidified by adding 2N aqueous sulfuric acid, the precipitated aluminum hydroxide partially dissolving. The (S) -4-bromo3-methyl-1-butanol formed is taken up in ether by repeated shaking. The combined ether phases are washed first with saturated sodium bicarbonate solution, then with saturated saline until neutral and dried over sodium sulfate.

  After removing the solvent on a rotary evaporator, the yield is 14.0 g (98%); the product can be used without distillation. For analytical purposes, a sample is distilled on the Kugelrohr at 600 / 0.04 Torr; [a] 20 = -2.00 (c = 3.3, CHCb).



   14 g (0.084 mol) of (S) -4-bromo-3-methyl-1-butanol are added dropwise at 00 with 50 ml of freshly distilled 3,4-dihydro-2H-pyran. The reaction mixture is then stirred at 0 for 1 hour. Excess 3,4-dihydro-2Hpyran is removed on a rotary evaporator at 35o. In order to remove the 3,4-dihydro-2Hpyran as completely as possible, chloroform is added repeatedly, in each case followed by distillation on a rotary evaporator. The crude product [(S) -4-bromo-3-methylbutoxyj-tetrahydro-2H-pyran is distilled at 75O / 0.03 torr. Yield; 17.5 g (83%) [a] D = + 3.40 (c = 4.0, CHCl3).



   In a 3-neck flask gassed with argon and equipped with a calcium chloride tube, 4.7 g (0.202 mol) of magnesium chips are activated by adding 2.0 ml of methyl iodide. After 5 minutes, the methyl iodide is suctioned off and the magnesium is washed several times with absolute tetrahydrofuran. A solution of 44 g (0.175 mol) of 2 - [(S) -4-bromo-3 methylbutoxy] tetrahydro-2H-pyran in 50 ml of absolute tetrahydrofuran is added dropwise to the activated magnesium at such a rate that the solvent is just boils. If the Grignard reaction does not start automatically, the oil bath is heated to 85 o.



  After the addition of 2 - [(S) -4-bromo-3-methylbutoxy] - tetrahydro-2H-pyran has ended, the mixture is stirred at 85 "(0 to 15 min.) Until, according to gas chromatographic analysis, only traces of 2- [(S) -4-bromo-3-methylbutoxy] tetrahydro-2Hpyran are present. The resulting solution is cooled to -780 and added dropwise with 21.2 g (0.0875 mol) of 3 methyl-1-butanol-p- toluenesulfonate, followed by 3.6 ml of a 0.1 molar solution of Li2CuCl4 in absolute tetrahydrofuran The reaction mixture is stirred at -780 for 10 min, then at 0 for 2 hours and then at room temperature for a further 14 hours.

  To work up the 2 [(R) -3,7-dimethyloctanoxy] tetrahydro-2H-pyran obtained, the mixture is poured onto ice and brought to pH 5 to 6 with 2N sulfuric acid. After extraction three times with ether and removal of the solvent on a rotary evaporator, a mixture of (R) -3,7-dimethyl-1-octanol and 2 [(R) -3, 7-dimethyloctanoxy] tetrahydro-2H-pyran is obtained. This mixture is mixed in portions in 100 ml of methanol at 0 with a total of 100 ml of 7N aqueous hydrochloric acid. After stirring for 2 hours at room temperature, the mixture is neutralized with dilute aqueous sodium hydroxide solution and extracted with ether.



  Chromatography on silica gel deactivated with 0.5% ammonia with pentane / ether (4: 1) gives 10.6 g (76% based on 3-methyl-1-butanol-p-toluenesulfonate) of pure (R) -3 , 7-dimethyl-1-octanoL bp 58-590 / 0.05 torr; [a] 20 = +4.0 (c = 1.03, CHCl3).



   The (R) -3,7-dimethyl-1-octanol can also be obtained as follows:
To a Grignard solution (prepared in analogy to the magnesium derivative of 2 - [(S) -4-bromo-3 methylbutoxyl-tetrahydro-2H-pyran) described above from 0.3 g (12.4 mmol) of magnesium and 1.51 g ( 10 mmol) of 1-bromo-3-methylbutane in 25 ml of absolute tetrahydrofuran, 1.26 g (5 mmol) of 2 - [(S) -4-bromo-3-methylbutoxy] tetrahydro at 00 under argon and with stirring -2H-pyrine added dropwise within one minute. Then 0.3 ml of a 0.1 m solution of Li2CuCl4 in tetrahydrofuran is added. The reaction mixture is continued at 00 for 3 hours.

  The working up and the hydrolysis of 2 - [(R) -3,7-dimethyl-octanoxy] -tetrahydro-2H-pyran to (R) -3,7-dimethyl-1-octanol is carried out in the manner indicated above. The yield of (R) -3,7-dimethyl-1octanol after Kugelrohr distillation at 950/14 Torr is 0.56 g (71%); [(k] 020 = +4.0 (c + 1.03, CHCl3).



   A solution of 6.5 g (41 mmol) of (R) -3,7-dimethyl-octanol and 7.8 g (41 mmol) of p-toluenesulfochloride in 50 ml of absolute chloroform is mixed at 0 "with 7.9 g (0 , 1 mol) of absolute pyridine is added, the mixture is then stirred for 20 hours at 40 ° C. For working up, the mixture is poured onto 500 g of ice and extracted three times with chloroform. The combined organic phases are washed first with cold in hydrochloric acid, then with saturated aqueous sodium bicarbonate solution and then After drying over potassium carbonate and removing the solvent under reduced pressure, the crude product is chromatographed on 300 g of silica gel with benzene.



  11.4 g (89%) of (R) -3,7-dimethyl-1-octanol ptoluenesulfonate are obtained; [a] 20 = +2.0 (c = 4.0, benzene).



   A Grignard solution of 2 g of magnesium and 18.3 g (72.6 mmol) of 2 - [(S) -4-bromo-3-methylbutoxyj-tetrahydro-2H-pyran in 50 ml abs. Tetrahydrofuran, prepared in the manner given below, is at -78 "with 11.4 g (36.3 mmol) of (R) -3,7-dimethyl-1-octanol-p-toluenesulfonate and 3 ml of a 0.1- molar solution of Li2CuCl4 in tetrahydrofuran added.



  Working up and hydrolysis of tetrahydro-2 {[(3R, 7R) -3,7, 1 1-trimethyldodecyl} -oxy} -2H-pyrans to (3R, 7R) -3,7,11-trimethyl-l- Dodecanol is carried out in analogy to the working up and hydrolysis of 2 [(R) -3,7-dimethyl-octanoxy] tetrahydro-2H-pyran described below. The (3R, 7R) -3,7,11-trimethyl-1-dodecanol distilled at 114 / 0.04 Torr with a yield of 7.0 g (84% based on (R) -3,7-dimethyl-1 octanol p-toluenesulfonate); [(t] o2t = + 3.7 O (c = 1.015, n-octane).



   The mixture of tetrahydro-2- {[(3R, 7R) -3,7,11-trimethyldodecyl] -oxy} -2H-pyran and (3R, 7R) -3,7,11-trimethyl- 1-dodecanol can be separated chromatographically [with silica gel deactivated by 0.5% aqueous ammonia; Eluent: toluene]. The tetrahydro-2- {[(3R, 7R) -3 7,11 -trimethyldodecyl] -oxy} -2H-pyran has an optical rotation of [cx] D = + 2.330 (c = 4.0, chloroform).



   The (3R, 7R) -3,7,11-trimethyl-1-dodecanol can also be prepared as follows:
To a solution of 5 g (31.6 mmol) of (R) -3,7-dimethyl-1octanol and 9.05 g (34.5 mmol) of triphenylphosphine in 20 ml of methylene chloride are added 5.65 g (31.8 mmol) N-Bromosuccinimide was added in portions with stirring. The temperature is kept below 25 ° by occasional cooling of the reaction vessel. After a stirring time of 30 minutes at room temperature, the solvent is removed on a rotary evaporator. The residue is washed out several times with n hexane, then filtered and washed again with n-hexane. The combined n-hexane phases are concentrated on a rotary evaporator. The crude product is chromatographed on 200 g of silica gel with n-hexane.

  Distillation in a bulb tube at 105/15 torr gives 6.1 g (87%) of (R) -1-bromo3,7-dimethyloctane; [(2t = -5.0 (c = 0.82, CHCl3).

 

   A Grignard solution of 0.3 g (12.4 mmol) of magnesium and 2.54 g (11.5 mmol) of (R) -1-bromo-3,7-dimethyloctane in 10 ml of absolute tetrahydrofuran is described in the above in Example 10 prepared manner. 1.45 g (5.57 mmol) of 2 - [(S) -4-bromo.3-methylbutoxy] tetrahydro-2H-pyran are added dropwise to this solution at 0O, followed by 0.3 ml of a 0 , 1 molar solution of Li2CuCl4 in tetrahydrofuran. The reaction mixture is stirred at 0 for 3 hours. Working up and hydrolysis of the tetrahydro-2 ([(3R, 7R) -3,7,11 -trimethyldodecyl] -oxy -2H-pyrans to (3R, 7R) -3,7,1 1-trimethyl-1- Dodecanol is carried out in the manner described in Example 10 above.

  The yield of (3R, 7R) 3,7,11 -trimethyl-1-dodecanol after Kugelrohr distillation at 90 to 950.05 Torr is 0.5 g (43% based on 2 - [(S) -4 bromo-3 -methylbutoxyj-tetrahydro-2H-pyran); [a] 200 = +3.9s (c = 1.05, n-octane).



   3.1 g (13.6 mmol) (3R, 7R) -3,7,11 -trimethyl-l-dodecanol are mixed with 4.02 g (15.3 mmol) N-bromosuccinimide and 2.62 g (14, 7 mmol) of triphenylphosphine in 13 ml of methylene chloride by the method described in Example 10 above. After chromatography and distillation, one obtains
3.54 g (90%) (3R, 7R) -bromo-3,7,11-trimethyldodecane. Sdp.



     90O / 0.05 Torr; [(t] 20 = -3.6 (c = 1.005, n-octane).



   The (3R, 7R) -1-bromo-3,7,1-trimethyldodecane obtained can be obtained according to Helv. Chem. Acta, Volume 46 (1963), pages 650
675 in (2R, 4'R, 82) - (t-tocopherol or (2R, 4'R, 8'R) -ct
Tocopheryl acetate can be converted.



   Example 12
250 ml of about 11 N ethanolic hydrochloric acid are added dropwise with 25.8 g of (S) -dihydro-3-methyl-2 (3H) -furanone within half a minute. The rapidly blackening solution is stirred at room temperature for 10 seconds and then extracted with methylene chloride. The organic phase is washed successively with saturated, aqueous bicarbonate solution and water, dried and under reduced pressure
Pressure reduced. After distilling the residue at 15
Torr gives 39.7 g (94%) of (S) -4-chloro-2-methyl-butteric acid ethyl ester as a colorless oil; Boiling point: 77-780. [(t] 20 = +26.90 (4.15% in ethanol).



   A solution of 4.9 g (30.0 mmol) of (S) -4-chloro-2-methyl butyric acid ethyl ester in 50 ml of n-hexane is removed at -70
Argon was added dropwise with 39 ml of a 20% solution of di-isobutylaluminum hydride in n-hexane. After finished
The reaction mixture is mixed with 5 ml of methanol at - 300, then hydrolyzed with ice-cooled 1N hydrochloric acid, extracted with ether, washed neutral with water, dried and evaporated under reduced pressure. (30 bath temperature). The residue is distilled at 55 to 58/15 torr. 2.3 g (64%) of (S) -4-chloro-2-methylbutyraldehyde are obtained as a colorless liquid, [ct] 20 = -33.4 "(4.21%, CHCl3).



   A solution of 72.0 g triphenylphosphine and 41.5 g
Isoamyl bromide in 250 ml of dimethylformamide is heated under reflux conditions for 2 hours. The solvent is distilled off under reduced pressure, the residue is crystallized twice from methanol / ether and finally dried for 48 hours at 110 ° under greatly reduced pressure. There remain 70.3 g (61.9%) of 3-methylbutyl triphenylphosphonium bromide, mp. 153 to 1540 (uncorrected).



   23.5 ml of n-butyllithium (about 2 m in n-hexane) are added dropwise to a suspension of 20.4 g of 3-methylbutyltrimethylphosphonium bromide in 80 ml of dry ether, with a reaction temperature of about 20 being maintained by gentle cooling . Then it is stirred for a further 2 hours at room temperature. The mixture is cooled to -10 ° and a solution of 5.4 g of (S) -4-chloro-2-methylbutyraldehyde in 5 ml of dry ether is added dropwise. The precipitate is stirred for one hour without a cooling bath, after which 100 ml of dimethylformamide are added dropwise at room temperature. The mixture is stirred at room temperature for 16 hours, hydrolyzed by adding ice and extracted with ether. The ether phase is washed with water, dried and concentrated under reduced pressure.



  The residue is purified by adsorption on silica gel and on neutral aluminum oxide of activities III and II (eluent: n-hexane). After distillation under reduced pressure (13 mm Torr / 90 "), 4.13 g (53%) (S) -cis 1-chloro-3,7-dimethyl-4-octene [(t] 20 = +32, 6 (2.15% in chloroform).



   A solution of 6.0 g of (S) -cis-1-chloro-3,7-dimethyl-4-octene and 10.8 g of triphenylphosphine in 40 ml of dimethylformamide is heated under reflux conditions for 20 hours. The mixture is concentrated under reduced pressure and washed with dry ether until only traces of triphenylphosphine are visible in the thin-layer chromatogram. The residue is dried under reduced pressure first at 50O and later under greatly reduced pressure at room temperature overnight. There remain 9.0 g (60.0%) of a light brown glassy mass. 20 ml of dry ether are added and 10.5 ml of 2 m n-butyllithium solution are slowly added dropwise.

  After stirring for a few hours under argon, a dark red color appears
Suspension formed, which is added dropwise at -10 with a solution of 2.45 g of (S) -4-chloro-2-methylbutyraldehyde in 3 ml of dry ether. The mixture is stirred for 1 hour without cooling, then 50 ml of dry dimethylformamide are added dropwise at room temperature and stirring is continued overnight.



  The reaction mixture is hydrolyzed by adding ice and extracted with ether. The extract is freed of dimethylformamide by washing, dried and evaporated under reduced pressure. The residue is purified by adsorption on silica gel (eluent: n-hexane) and distilled in a Kugelrohr (85o / 0.05 Torr. This gives (3S, 7S) -1-chloro-3,7,11-trimethyl-4-cis -trans-8-cis-dodecadiene. Yield 2.7 g (55%). [ct] o20 = -6.5o (4.07% in CHCl3).



  The cis / trans ratio and thus the rotation value varies from batch to batch.



   The compound obtained is used to prove the optical purity of prehydrogenated platinum dioxide to (3R, 7R) -1-chlorine
3,7,11 -trimethyldodecane hydrogenated; Boiling point 90 / 0.07 torr.



     [(ti205 = -1.20 (4.08% in CHCl3). The hydrogenation product is converted into the corresponding ester by heating with dry potassium acetate in dimethylformamide (4 hours at 170). The ester is adsorbed on silica gel (eluent: n -Hexane / ether 4 + 1), then further converted to (3R, 7R) -3,7,1 1-trimethyl-1-dodecanol by stirring with 4N aqueous potassium hydroxide solution (30 min. At room temperature).

  It is extracted, washed neutral, dried, concentrated, distilled in a bulb tube (1150 / 0.20 torr) and obtained (3R, 7R) 3,7,11-trimethyl-1-dodecanol [yield 94% based on (3R, 7R) -1-chloro-3,7,11-trimethyl-dodecane]. The product has an optical rotation value of [U] 20 = + 3.76 (4.14% in n-octane) and is therefore identical to authentic material.



   The (3S, 7S) -1-chloro-3,7,11-trimethyl-4-cis / trans-8-cisdodecadiene can also be prepared as follows:
A solution of 4.4 g of (S) -1-chloro-3,7-dimethyl-4-octene in 30 ml of isobutyl methyl ketone is stirred with 7.55 g of sodium iodide at 130 for 37 hours. Another 3.75 g of sodium iodide are added and stirring is continued for 4 hours. After cooling to room temperature, the mixture is diluted with ether, washed with water, dried and concentrated under reduced pressure. The residue is pre-cleaned by adsorption on silica gel (eluent: n-hexane). The pure fractions are combined and distilled at 1300/14 torr. 5.6 g (83.5%) of (S) -cis-1-iodo-3,7-dimethyl-4-octene are obtained as a colorless liquid. [U] DO = + 14.30 (2.21% in CHCl3).

 

   A solution of 6.1 g of (S) -cis-1-iodo-3, 7-dimethyl-4-octene and 7.2 g of triphenylphosphine in 40 ml of toluene is kept under reflux for 24 hours, then concentrated under reduced pressure. The residue is digested with absolute ether until only traces of triphenylphosphine are visible in the thin layer chromatogram. 11.8 g (97.4%) of (S) -cis-3,7-dimethyl-4-octene-1-triphenylphosphonium iodide are obtained. This is overlaid with 20 ml of dry ether and 22.2 ml of a 2 m solution of n-butyllithium in n-hexane are added dropwise in an argon atmosphere with stirring and gentle cooling to room temperature.

  The mixture is stirred at room temperature for 20 seconds, cooled to -10 and a solution of 2.8 g of (S) -4-chloro-2-methylbutyraldehyde in 3 ml of dry ether is added dropwise. The mixture is stirred for 1 hour without cooling, concentrated to a small volume under reduced pressure, 80 ml of dimethylformamide are added to the residue while cooling to room temperature, and the solution is then stirred at room temperature for 20 hours. The reaction mixture is poured onto ice, extracted with ether, freed from traces of dimethylformamide by washing with water, dried and concentrated under reduced pressure.



  The residue is purified by adsorption on silica gel (eluent: n-hexane) and distilled at 1000 / 0.2 torr.



  (3S, 7S) -1-chloro-3,7,11-trimethyl-4-cis / trans-8-cisdodecadiene is obtained in a yield of 1.7 g (30.1%). [a] 20 = 5.2 (2.11% in chloroform).



   2.0 g (3S, 7S) -1-chloro-3,7, 1 1-trimethyl-4-cis / trans-8-cis-dodecadiene are mixed with 2.5 g triphenylphosphine in 20 ml dry dimethylformamide under reflux conditions for 24 hours heated. The solvent is removed under reduced pressure and the residue is digested with dry ether until only traces of triphenylphosphine are visible in the thin layer chromatogram. The residue dried in a fine vacuum weighs 1.85 g (44.5% yield) and consists of (3S, 7S) -3,7,11-trimethyl-4-cis / trans-8-cis-dodecadiene-1-triphenylphosphonium chloride. 10 ml of dry ether are then added under argon, followed by 1.8 ml of 2 m of n-butyllithium in n-hexane.

  A brown-red suspension is formed after 5 hours. At -10, a solution of 900 mg (S) -6-acetoxy-2-formyl-2,5,7,8-tetramethylchroman in 5 ml of dry ether is added dropwise in 15 minutes, the mixture is stirred for 1 hour without cooling and mixed then with 30 ml of dry dimethylformamide. The mixture is stirred for a further 18 hours at room temperature, hydrolyzed with ice, extracted with ether after saturation with sodium chloride, washed free of dimethylformamide, dried and concentrated under reduced pressure. It is purified by adsorption on silica gel, concentrated and the residue is reacted in 2 ml of dry pyridine with 2 ml of acetic anhydride (17 hours at room temperature). The reaction product is hydrolyzed with 3N hydrochloric acid, water and aqueous sodium bicarbonate solution.

  After drying and concentrating under reduced pressure, the residue is purified by adsorption on silica gel (eluent: chloroform), concentrated under reduced pressure and dried to constant weight under greatly reduced pressure. 1.29 g (75%) (2S) -2,5,7,8-tetramethyl) -2 - [(4S, 8S) -4,8,12-trimethyl-1-cis / trans-5 are obtained -cis / trans-9-cis-tridecatrienyl] -6-chromanyl acetate as a pale yellow, viscous oil; [ttio2t = -60.7 "(2.11% in CHCl3).



   The product is a cis / trans isomer mixture, which has a different composition in each batch and also has different rotation values.



   A solution of 321 mg (2S) -2,5,7,8-tetramethyl-2 [(4S, 8S) -4, 8,12-trimethyl-1-cis / trans-5-cis / trans-9-cis -tridecatrienyl] -6-chromanyl acetate in 5 ml of pure ethyl acetate is hydrogenated with the help of 35 mg of hydrogenated platinum dioxide. After 25 minutes, 60.0 ml of hydrogen are taken up. The catalyst is filtered off, the filtrate is concentrated under reduced pressure and the residue is dried under greatly reduced pressure. 319 mg (98.1% (2R, 4'R, 8iR) - (t-tocopheryl acetate as a colorless, viscous oil [] DO = +2.20 (2.09% in cyclohexane), identical to authentic material.



   Starting from (3S, 7S) -1-chloro-3,7,11-trimethyl-4-cis / trans-8-cis-dodecadiene, one can (2R, 4'R, 8'R) -cr-tocopheryl acetate also win in the following ways:
A solution of 1.6 g (3S, 7S) -1-chloro-3,7,11-dimethyl-4cis / trans-8-cis-dodecadiene in 20 ml of isobutyl methyl ketone is mixed with 3.0 g of sodium iodide for 48 hours at 1300 ( Bath temperature) stirred. The reaction mixture is cooled to room temperature, taken up in ether, washed with water and evaporated under reduced pressure. The residue is purified by adsorption on silica gel (eluent: n-hexane).

  After distillation at 100o / 0.07 Torr, 1.98 g (90%) (3S, 7S) -1-iodo-3,7, 1 1-trimethyl-4-cis / trans-8-cis-dodecadiene is obtained colorless product; [a] D = -7.3O (4.11% in chloroform).



   The product is a cis / trans isomer mixture, which has a different composition in each batch and also has different rotation values.



   The optical purity of the product obtained is checked as follows:
It is hydrogenated on prehydrogenated platinum oxide and the iodine atom is exchanged for the hydroxyl group by heating with potassium acetate in dimethylformamide and saponifying with 4N aqueous sodium hydroxide solution. The resulting product, (3R, 7R) 3,7,11-trimethyl-1-dodecanol, has a rotation of [(4025 = +40 (4.14% in n-octane) and is therefore identical to authentic, optically pure Material.



   A solution of 1.1 g (3S, 7S) -1-iodo-3,7'11-trimethyl-4 cis / trans-8-cis-dodecadiene, 1.0 g triphenylphosphine and 10 ml xylene is refluxed for 24 hours heated, then concentrated under reduced pressure. The residue is digested with dry ether until only traces of triphenylphosphine are visible in the thin-layer chromatogram. The dried residue consists of (3S, 7S) -3,7,11trimethyl-4-cis / trans-8-cis-dodecadien-1-triphenylphosphonium iodide and weighs 1.9 g (97%).



   The phosphonium salt is overlaid with 10 ml of dry ether and 1.6 ml of 2 m of n-butyllithium in n-hexane are added dropwise and the mixture is stirred at room temperature for 2 hours. At -15, a solution of 880 ml (S) -6 acetoxy-2-formyl-2,5, 7,8-tetramethyl-chroman in 10 ml dry ether is added dropwise in 15 minutes, the mixture is stirred for 1 hour without cooling and mixed then with 30 ml of dry dimethylformamide. The mixture is stirred for 20 hours at room temperature, then hydrolyzed by adding ice and extracted with ether after saturation with sodium chloride. The ether extract is washed with water, dried and concentrated under reduced pressure.

  It is purified by adsorption on silica gel (eluent: chloroform), concentrated and the residue is re-acetylated in 2 ml of dry pyridine with 2 ml of acetic anhydride (17 hours at room temperature). The product obtained is hydrolyzed with ice, extracted with ether and the ether phase is washed with 3N hydrochloric acid, water and aqueous sodium bicarbonate solution. After drying and concentrating under reduced pressure, the residue is purified again by adsorption on silica gel (eluent: chloroform), concentrated under reduced pressure and then dried to constant weight under greatly reduced pressure.

  459 mg (31%) (2S) -2,5,7,8-tetramethyl-2 [(4S, 8S) -4,8,12-trimethyl-1-cis / trans-5 -cis / trans- are obtained. 9-cis-tridecatrienyl] -6-chromanyl acetate as a pale yellowish oil. [] 20 = -41.30 (0.75% in CHCb).



   The product is a cis / trans isomer mixture, which has a different composition in each batch and also has different rotation values.

 

   A solution of 400 mg (2S) -2,5,7,8-tetramethyl-2 [(4S, 8S) -4,8,12-trimethyl-I-cis / trans-5-cis / trans-9-cis -tridecatrienyl] -6-chromanyl acetate in 5 ml of pure ethyl acetate is hydrogenated with the help of 50 mg of prehydrogenated platinum dioxide. After 40 minutes, 68 ml of hydrogen are taken up. The catalyst is filtered off, the filtrate is concentrated under reduced pressure and the residue is then dried under greatly reduced pressure. 396 mg (97.7%) (2R, 4'R, 8'R) - (t-tocopheryl acetate in 99.7% purity are obtained. [(T] 20 = + 2.20 + 0.5O (1, 07% in cyclohexane), identical to authentic (2R, 4'R, 8'R) - tt - tocopheryl acetate.



   Example 13
300 mg (2.3 mmol) of ethyl acetoacetate and then 671 mg (2.3 mmol) (3R, 7R) are added dropwise to 2.3 ml of a solution of 1.01N sodium ethylate in ethanol in a flask equipped with a reflux condenser and calcium chloride tube. -1-bromo-3,7,11-trimethyldodecane in 2 ml of ethanol.



  The mixture is heated to boiling under reflux conditions for 15 hours with stirring. After cooling, add enough ice water to dissolve the separated salt, separate the organic phase in a separatory funnel and shake it four times with methylene chloride. After drying the combined organic phases over magnesium sulfate and distilling off the solvent, 755 mg of crude (5R, 9R) -2-acetyl-5,9,13-trimethyl-tetradecanoic acid ethyl ester are obtained.

  For analytical purposes, a sample of the crude (5R, 9R) -2-acetyl-5,9,13-trimethyl-tetradecanoic acid ethyl ester is chromatographed on a preparative silica gel plate with toluene / hexane / ethyl acetate = 30: 10: 3 and the resultant is obtained pure (5R, 9R) -2-acetyl-5,9,13-trimethyl-tetradecanoic acid ethyl ester distilled at 150 / 0.03 torr; [(t] 36 5 - = -4.00 (c = 1.04; CHCl3).



   A solution of 154 mg (5R, 9R) -2-acetyl-5,9,13-trimethyl-tetradecanoic acid ethyl ester in 5 ml of ethanol is added dropwise at a temperature of 80 with 1 ml of 10% sodium hydroxide solution. The reaction mixture is then heated to boiling under reflux conditions for 3.5 hours. The cold reaction mixture is neutralized with aqueous hydrochloric acid and extracted four times with methylene chloride. The combined organic phases are first washed with saturated saline and then dried over magnesium sulfate.

  After removal of the solvent under reduced pressure and distillation at 1500 / 0.03 torr, 118 mg (95%) (6R, 10R) - (6R, 10R) -14-trimethylpentadecanone-2 (hexahydrofarnesylacetone) are obtained. [(Liit3s = +7.55 (c = 1.57; n-octane). The ORD spectrum is identical to that of (6R, IOR) -14-trimethylpentadecanon-2 made from natural phytol (Helv. Chem. Acta , 47, 1964, pages 221 ff).



   The (6R, 1OR) -14-trimethylpentadecanone-2 obtained can e.g. according to J. Chem. Soc. (C), 1966, pages 2144-2176 and



  Helv. Chim. Acta, 48, 1965 pages 1332-1347 can be converted into natural vitamin Kl.


    

Claims (10)

    PATENT REQUESTS 1. Process for the preparation of tertiary optically active aliphatic compounds of the general formula EMI1.1  in which one of the radicals X 'and Y' represents optionally esterified carboxy and the other represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl, where X 'and Y' have different functional meanings and furthermore free carboxy and hydroxymethyl groups with one another to form the group -CO-O-CH2- or -CH2-O-CO- are lactonized, and if X 'represents alkyl-etherified hydroxymethyl, Y' can also optionally be esterified hydroxymethyl, characterized in that an aerobic or facultative aerobic microorganism containing one in hydrogenates an aliphatic chain between CH and methylated C double bond,  to a compound of the general formula EMI1.2  in which X represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl and Y optionally esterified carboxy, optionally acetalized formyl or optionally etherified or esterified hydroxymethyl, where X and Y have different functional meanings and furthermore, if X is optionally esterified hydroxymethyl, Y is esterified Represents carboxy or optionally acetalized formyl and, if Y is optionally esterified hydroxymethyl, X is esterified carboxy or alkyl-etherified hydroxymethyl, can act in an aqueous medium.  2. The method according to claim 1, characterized in that one carries out the fermentative hydrogenation under aerobic conditions with a microorganism in a non-growing (stationary) phase.  3. The method according to claim 1 or 2, characterized in that one carries out the fermentative hydrogenation in the presence of an assimilable carbon source.  4. The method according to claim 3, characterized in that a sugar is used as the carbon source in an amount of about 10 to 100 g per liter of aqueous medium.  5. The method according to any one of claims 1 to 4, characterized in that one carries out the fermentative hydrogenation at a pH of 2 to 10, in particular 3 to 8.  6. The method according to any one of claims 1 to 5, characterized in that one carries out the fermentative hydrogenation at a temperature of 20 to 35 C.  7. The method according to any one of claims 1 to 6, characterized in that the starting material of formula I in a concentration of 0.1 to 5.0%, in particular 0.5 to 2.5%, based on the aqueous medium, starts.  8. The method according to any one of claims 1 to 7, characterized in that one uses a starting compound of formula I in which X is optionally esterified carboxy and Y optionally acetalized formyl or X is esterified carboxy and Y is optionally etherified or esterified hydroxymethyl, and as a microorganism Saccharomyces cerevisiae (press yeast; baker's yeast) is used to obtain a fermentation product of the formula II in which X 'represents optionally esterified carboxy and Y / optionally etherified or esterified hydroxymethyl, free carboxy and hydroxymethyl groups being formed together to form the group -CO-O -CH2- are lactonized.  9. The method according to claim 8, characterized in that one uses a starting compound of the formula I in which X is optionally esterified carboxy and Y is optionally acetalized formyl.  10. The method according to claim 9, characterized in that the starting compound of the formula I is ethyl trans4,4-dimethoxy-3-methylcrotonate, the main fermentation product of the formula II being (S) -3 methyl-4-hydroxy- Obtains ethyl butyrate, which is converted into (S) -dihydro-4-methyl-2 (3H) -furanone by acid hydrolysis.  The invention relates to a process for the preparation of tertiary, optically active aliphatic compounds of the general formula EMI1.3  in which one of the radicals X 'and Y' represents optionally esterified carboxy and the other represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl, where X 'and Y' have different functional meanings and furthermore free carboxy and hydroxymethyl groups with one another to form the group -COSCH2- or -CH2-OCO are lactonized, and if X 'represents alkyl-etherified hydroxymethyl, Y' can also optionally be esterified hydroxymethyl.  The process according to the invention is characterized in that an aerobic or facultative aerobic microorganism which hydrogenates a double bond located in an aliphatic chain between CH and methylated C, on a compound of the general formula EMI1.4  in which X represents optionally esterified carboxy or optionally etherified or esterified hydroxymethyl and Y optionally esterified carboxy, optionally acetalized formyl or optionally etherified or esterified hydroxymethyl, where X and Y have different functional meanings and furthermore, if X is optionally esterified hydroxymethyl, Y is esterified Represents carboxy or optionally acetalized formyl and if Y optionally represents esterified hydroxymethyl, X represents esterified carboxy or alkyl-etherified hydroxymethyl,  can act in an aqueous medium.  The process according to the invention opens up a new advantageous route to intermediates in the synthesis of natural, optically active vitamin E, vitamin E ester and vitamin Kl, compounds which previously had to be obtained in an uneconomical manner from natural products or by complex synthetic methods. Because of the new method according to the invention, new, optically active compounds of the formula II are obtained in a particularly simple manner, which are advantageous to the ** WARNING ** End of CLMS field could overlap beginning of DESC **.