40.50
COMMUNICATIONS TO THE EDITOR
VOl. 70
rium is involved between a carbonyl of cobalt (possibly dicobalt octacarbonyl or the hydrocarbonyl), carbon monoxide and the olefin, the latter being able to substitute for some of the CO in the carbonyl. One possible scheme for the reaction is
vapors yielded values within estimated error of the atomic weights. The existence of polyatomic ionic species in the vapor of silver metal was recently established through direct observation of the ions in a 60" single direction focusing mass spectrometer of very high [ c o ( c o h l z f CsHio [COs(CO);.CsHiol f CO (1) sensitivity. I n one run a t various temperatures (A) above the melting point and below 1500" silver va2CO(CO);.CsHio f 2H2 ----f por was found to contain ionic species (presumably ~ C G H I I C H O [cO(co)~]4 (2) formed by thermal ionization) corresponding to the [Co(CO),], f 4CO If 2[C0(cO),]s (3) compositions Agf, Agz+, Ag3+ and Agdf in approxiI t is possible that the postulated intermediate mate relative intensities 15:7 : 30 : 1, respectively. Identification of these species was made on the (A) is formed from the decomposition of the complex [Co(C0)4.0lefin]+[Co(C0)4]- recently sug- basis of the mass numbers observed, which were gested by Orchin3 but further work is required, known from magnetic field calibration with a proespecially since the above tentative scheme does ton resonance fluxmeter, and on the basis of a characteristic mass spectrum for each species which renot assign a role to the hydrocarbonyl. sults from the isotopic composition of silver. It (3) M. Orchin, in "Advances in Catalysis," Vol. V, Academic Press, has not been possible as yet to search for higher Inc., New York, 1953, p 407. polymers which may also exist. DEPARTMENT OF INDUSTRIAL CHEMISTRY G. SATTA There is, of course, considerable uncertainty asPOLYTECHNIC INSTITUTE R. ERCOLI OF MILAS S. CASTELLANOsociated with the extension of these data to the ITALY F. H. BARBIERI relative abundances of neutral polyatomic species in RECEIVEDMAY3, 1954 the vapor state, but it may be noted that the above relative intensities would give an average molecular weight for silver vapor in good agreement with the POLYATOMTC SPECIES IN SILVER VAPOR' above molecular weight determination.
+
Sir :
Calculation of the molecular weight of silver vapor from the measured vapor velocity2 and success of the famous Stern-Gerlach experiment in yielding in an inhomogeneous field the splitting predicted for a beam of silver atoms3 apparently established beyond doubt that silver vapor is monoatomic a t low pressures. We have found, however, a high concentration of polyatomic species in silver vapor a t -IOp6 to l o p 4atm. pressure. By use of a method developed by Mayer4 and V ~ l m e rwe , ~ have measured the apparent molecular weight of silver vapor in a graphite box containing two small holes. Pressure inside the box can be calculated from the hole areas, rate of weight loss due to escaping vapor, temperature and the molecular weight of the vapor by use of Knudsen's equations6 The pressure can also be calculated from the angle 4 through which the force of the escaping vapor causes the box to rotate about a wire of torsion constant D. The equation used is P = 2D$/(qlal q2a2),where al and a2 are the areas of the holes and q1 and q 2 are distances from the axes of the holes to the wire. Simultaneous solution of the equations yields the molecular weight M of the vapor. Fifteen determinations a t 1310 to 1420°K. gave M = 278 (2.56 times the atomic weight) with an estimated maximum uncertainty of f90. This high average molecular weight cannot arise from systematic error because concurrent determinations of the molecular weights of tin' and germanium8
+
(1) Supported in part by the Office of Naval Research and in part by the Atomic Energy Commission. (2) 0. Stern. 2 . Physik, 3 , 417 (1920). (3) W. Gerlach and 0. Stern, ibid., 9, 319 (1922). (4) H. Mayer, ibid., 67,240 (1931). (5) hl. Volmer, i b i d . , Chem., Bodenstein Feslband, 863 (1931). (6) M. Rnudsen, A n n . Physik, [4] 28, 999 (1909). (7) A. W. Searcy and R . D. Freeman, TH!S JOURNAL, in press. (8) A. W. Seurcy and R,D, Freeman, tlnpuhlished dntn.
DEPARTMENT OF CHEMISTRY ALANW. SEARCY ROBERTD. FREEMAN PURDUE UNIVERSITY WESTLAFAYETTE, INDIANA RADIATIOX LABORATORY MAYNARD C. MICHEL UNIVERSITY OF CALIFORNIA BERKELEY, CALIFORNIA RECEIVED JULY 8, 1954 MICROBIOLOGICAL TRANSFORMATIONS OF STEROIDS. XII.' 17a-HYDROXYLATION
Sir: Preceding publications from our own as well as other laboratories have been concerned with the microbiological oxygenation of the steroid molecule a t various positions of the skeleton. The introduction of a hydroxyl group into positions 6/3,2.3 7 / 3 , 2 , 4 g(,Z 1 1 a , 1 , 2 , 3 , f i , 6 11/3,2.7,8,9 1 4 a , 2 , 1 0 15," and 16a,12*13 by the action of various genera of the filamentous fungi has been described. In some instances a simultaneous introduction of two hy(1) Paper XI of this series, THISJ O U R N A L , in press. (2) ~, H. C. Murrav a n d D. H. Peterson, U. S. Patent 2,602,709 (July 8, 1952). (3) Papers 111, IV, V, IX, and X of this series, THISJ O U R N A L , 7 6 , 408,412, 416, 5768 (1953); 76,3174 (1954). (1) F. W. Kahnt, e l d . ,Ezpeuicnfia, 8, 322 (1952). ( 5 ) See, besides ref. 3, also papers I, 11, VI and VI1 of this series, THISJ O U R N A L , 74, 5933 (1952); 76, 55,419, 421 (1953). (6) 1. Flied, e t at., ibid., 74, 3962 (1952). (7) D. R. Colingsworth, et al., ibid., 74, 2381 (1952); J . Biol. Chem., 203, 807 (1953). (8) F . R. Hanson, e t al., TAISJ O U R N A L , 76, 5369 (1953). (9) G. M. Shull, D. A. Kita and J. W. Davisson, U. S. Patent 2,G58,023 (Nov. 3, 1953). (10) P. D. Meister, el a l . , Abstracts of the 123rd hleeting of the American Chemical Society, Los Angeles, Calif., March 15-19 (19531, page 5C. (11) (a) Personal communication from Dr. J. Fried, T h e Squibb Institute for Medical Research, New Brunswick, N. J.; (b) u n p u b lished results, these laboratories. (12) D. Perlman, E. Titus and J. Fried, THISJ O U R N A L , 74, 2126 (1952). (13) E, Vischer, J. Schrnidlin and A. Wettstcin. Helv. Chim. Acta. 87, 821 11854).
Aug. 5, 1954
COMMUNICATIONS TO THE EDITOR
4051
droxyl groups into the substrate has been ob- Pratt, Jennie I. Mejeur and Henrietta Triemstra served.1~2~5~6~12 However, dihydroxylated products and Mr. J. R. Heald and G. Staffen for technical have been formed to any appreciable extent only assistance, to Dr. J. L. Johnson and associates for the spectra and to Mr. W. A. Struck and associwith p r o g e ~ t e r o n e ~or * ~ 3-ketobisnor-4-cholen.~~~~ ates for rotations and microanalyses. 22-all as a substrate. Since thus far no 17-hydroxylation by microbioP. D. MEISTER L. M. REINEKE logical means has been reported, i t is the purpose of LABORATORIES R. C. MEEKS this communication to describe the enzymatic RESEARCH H. C. MURRAY THEUPJOHNCOMPANY oxygenation of the 17a-position of Czl-steroids by KALAMAZOO, MICHIGAN S. H. EPPSTEIN Cephalothecium roseum Cda (A.T.C.C. 8685). Upon H. M. LEIGHOSBORN incubation of 11-desoxysteroids with C. roseum, A. WEINTRAUB D. H. PETERSON the 17a-hydroxyl group was introduced with or RECEIVED JULY 6, 1954 without concomitant hydroxylation of the Sp- or the 1la-position. When 11-keto or 1lp-hydroxy17-desoxysteroids were used as substrates only 17THE STEREOCHEMISTRY OF METAL hydroxylation took place. CHELATES OF A POLYDENTATE LIGAND When desoxycorticosterone was incubated with Sir: a 48-hour growth of C. roseum on a corn steep-gluMetal complexes with sexadentate ligands were cose medium for 48 hours, extraction of the fermentation liquor with methylene dichloride and chroma- first described by Dwyer and Li0ns.l Diehl and tography of the concentrates over Florisi13 yielded co-workers2 seemingly prepared some similar comtwo products: (a) 1la,l7a,21-Trihydroxy-4-preg-pounds with triethylenetetramine and salicylaldenene-3,2O-dione2t4t6+l4 (11-epi F). The identity of hyde (also substituted salicylaldehydes) by the this compound was established through these physi- reaction of cobalt salts during their investigation on cal constants: m.p. 206-211 O, m.p. in admixture with the oxygen-carrying synthetic chelates. They, howauthentic 11-epi F, 208-212'; [ a ] =+121" (metha- ever, did not isolate the pure compounds or study nol); ( [ a ] =+117O in methanol for authentic 11-epi their properties. A Schiff base (I)from triethyleneF); the infrared spectrum of the diacetate (in HOC6HaCHN(CH2)zNCHzCHzN(CHz)&:=CHCeH4OH chloroform solution) was identical to the spectrum \CH/ of the authentic 11-epi F diacetate. (b) 6P,17a,I 1 21-Trihydro~y-4-pregnene-3,20-dione~~~~~~ was also CsH4OH isolated; m.p. 234-236', [ a ] f~5 3 " ; A&. 238mp, E 12,900. Anal. Calcd. for CalHaoOs: C, 69.58; tetramine (11) and salicylaldehyde (111) has been ,~ gives a brown hyH, 8.34. Found: C, 69.58; H, 8.38. The infrared reported by M ~ k h e r j e e which spectrum of the diacetate, m.p. 190-193', was iden- groscopic substance with [Co(py)*Cl*]CI. This is soluble in water, insoluble in alcohol, and decomtical to the spectrum of an authentic specimen. Paper chromatography indicated that small poses a t 110'. The same substance is also said to amounts of Reichstein's compound S and epicorti- be obtained by the reaction of (111) on [Co trien C12]Cl. costerone had been formed. The nitrogen analog of Dwyer's Schiff base (IV) Fermentation of the following substrates with C. roseum gave the corresponding 17a-hydroxy HOCsH4CH=N( CHz)zNH(CHz)zNH(CHz)zN=CHCaH4OH derivatives : could not be prepared from (11) and (111),only (I) being obtained. We have, however, been able to Substrate Conversion Product Progesterone llor,l7or - Dihydroxyprogest- prepare a large number of compounds of the sexadentate ligand (IV) by the action of (11)on bis-salierone cylaldehyde compounds of cobalt, copper and other 11-Dehydrocorticosterone Kendall's Compound E Corticosterone Kendall's Compounds F and bivalent metals in about 80-85% yield. These can be prepared also by adding a mixture of (11) E and (111) in four equivalents of alkali (KOH) in a It is worthy of note that the enzyme systems of cold methanol solution to a solution of the metal salt Cephalothecium can introduce a hydroxyl group a t concerned. (I) also gives a poor yield (10-20%) carbon atom 17 without any interference from a of these compounds by careful manipulation. A hydroxyl group already present a t carbon atom 21 compound of the composition [Co trien CHOCa(as in desoxycorticosterone). This is in contrast t o H40]++ is obtained as the main product from [Co the performance of the mammalian adrenal which, trien C12]Cland (111). a t least under the in vitro conditions employed by When base (I) reacts with metal salts, one molethe Worcester group, l6 cannot oxygenate the 17- cule of salicylaldehyde is hydrolyzed from it, and position of desoxycorticosterone. derivatives of base (IV) result. For bivalent metal The authors are indebted to Misses Irene N. ions such as Fe'I and PdlI these have the composition intermediate between MTS2 and MTS3, (14) For chemical syntheses of this compound or its diacetate, cf. J. Romo, et al., Chcm. and Ind.. 783 (1952); H . L. Herzog, et al., THIS where TSP and TS3 represent the bi-negative ions JOURNAL, 74, 4470 (1952); A. Lardon a n d T. Reichstein, Phorm. of the bases (IV) and (I), respectively. Acta Hclu., 47, 287 (1952); J. Romo, e1 al., THISJOURNAL, 76, 1277 The complexes of (IV) will be asymmetric (1953). (16) For a chemical synthesis of this compound cf. I(.Florey e n d M. Ehrenstein, J . Org. Ckcm., in press. (In) H.T.evu, el a t , , I,Blal. Chrm., 108,4118 (196S),
(1) F. P. Dwyer a n d F. Lions, THISJOURNAL, 69,2917 (1947). (2) H. Diehl a n d co-workers, Iowa Slat# COIL1.Sci., 91, 108 (1947). (8) A. K.Mukbcrjee, Scicncr nnd CuNwrr, 107 (186s).
le,
