Relative and absolute stereochemistry of diols obtained from microbial oxidation of 3-methylcyclohexene

Printed in Qreat Brihin.

TetrahedronLetter4 No. 25, pp 2137 - 2138, 1975. Dergttmon Prom.

RGATIVEANDABSOLUTESTEREtXIiEHIS~

OFDIOLS OBTAINEDFROHlUCROBIALOXIDATION

ol?3&ErEUfcUZExENE Herman Zlffer* and D.T. Gib4on+ *NationalInstituteof Arthritis, Netabolkm, and Mge4tive Mseasee, National IMtituk4 of Heelth, Bethe4da,Maryland 20014 end +Universityof Tern,, Au4tin, Texae (Receivedin USA 15 April 1975; receivedin UK for publication6 May 1975) Althou& a considerableliteraturi, exi4ts on the microbialoxidationof aliphaticend oomparativelyfew 4tudie4 have exaednedmicrobialoxidation4of aromatio compotlnd4*(') 4imple alicyellchydrooerboas.(2)In order to determinethe stereoohemi4tryof the trewformationof a carbon-carbondouble bond into a formedby

the acti@nof amutant

glycol, we exemlned the product4

rici.~Z

of Peeudomonaeputida (P. putlda39/D) on(i) 3-

etr4in

methylcyolohexene.A portion of the crude oxidationproduct4obtainedwith P. putida 39/D grown in the preeenceof (2) 3-methylcyclohexene wae convertedto bill-trlmethybilyl ethers, with hexamethyldi4ilazauein p@dine, where authentic4aaple4

and examinedby r.p.c. under conditions(4ee Table I)

(3) of the four immerlc

1,2-dihydro~-3-methylcyelohexane4 oouldbe

4eparated. The only volatile compound4observedunder the4e oonditioxwwere Thor of the bi4-trimethyl4ilylether4of the -ii4

diol4~and2&

TablO c.f.3. ha4d4

of m0lrr aa TrimethylSi41

ow tran4, --

1.25 a4oertain

whether

one

or

dized, the crude dial fractionwae reacted and the mixture (lb, 2b, &and21 va4 confirmedby i4olatlonof 2

tran4

Bi

(24

1.34

tran4

1.77

both lnantiomer4of )_wthylayolohexenewere odwith one equivalentof 3&dini~benzoyl

separated. The and g

T-75'; I$ W-1 Colporrnd

1.00

In order to

Ethar4

Relative retentiontime (R,)

Ccapopnd

end

analy4i4

4tereoche4l4try

of their

a44igmd

to the dio3s

mu 4pectra. Each compound

wa4 opticallyactive, e4tablkshingthat P. putida ~44 etereo4electivein ox%dia lnzutiomer4. In order

chloride

both

to determinewhether one or both enantiomer4were oxidized the

lute 4tereoehemi4tryof each diol VU determinedwing a combinationof ehemlcaland

abrro-

plty4ical

uthod4.

The

dlol&i (aD31.7Oi CRC+

orI.

prepzredby

hydroly4i4

of 2

uaa convertedto the di-

~~~~o,~~~~lchloridminplri~~~ea.d.~eoi~~~d~~ oak

dotemined.

222m.

The c.d. curve exhibitedanegati~e band at237 naud

We have 4howJ4) larUer that@&

ad.

&tehof&

2137

requir44

apo4ltive band at the

ab4dute

4ter4o-

2138

No. 25

R

CH

CH3ee

&

0 R'

(a)

(a) R - R' - H

1

R - R' = H

(b) R = H.

2

(b)R- H,

R' - 3,%dinitrobenzoyl (c) R- R' - benzoyl

= 3,5-dinitrobenzoyl (c) :' - 3,5-dinitrobenzoyl R' = H (d) R = R' = 3,5-dinitrobenzoyl

chemistry be to 3(R)-methyl-l(S),2(R)-dihydroxycyclohexane. The diol -2a (aD-35.2', CHC13, c=1.2) was prepared by hydrolysis of -2d and the absolute stereochemistry

of the methyl group established by oxidation to (+)-2(S)-methyl adipic acid

of known absolute stereochemistry. (5)

This result, in conjunction with the known relative

stereochemistry of Za, establishes -2a as 3(S)-methyl-l(S),2(R)-dihydroxycyclohexane. The observation that the absolute stereochemistry at the carbons bearing hydroxyl groups in each case is l(S),Z(R) also suggests that either the same or a very similar enzyme is involved.

These results represent one of the few authenticated

cases where microbial oxi-

dation of an olefin results exclusively in the formation of a -cis diol. REFERENCES (1)

D. T. Gibson, -Crit. Revs. Microbial. 1, 199 (1971).

(2)

B. B. Mukherjer, G. Kraidman and I. D. Hill, Applied Microb. 25, 447 (1973).

(3)

H. Ziffer, J. I. Seeman, R. J. Highet

(4)

H. Ziffer, D. M. Jerina, D. T. Gibson and V. M. Kobal, 2. ---Am. Chem. Sot. 95, 4048

and E. A. Sokoloski, J. Q.

(1973). (5)

C. F. Wong, E. Auer and R. T. LaLonde, 2. s.

Chem. 35, 517 (1970).

Chem. in press.