COMMUNICATIONS TO THE EDITOR
Sept., 1949
is correct, and if the molecular configuration of the tetramethylplatinum tetramer is unaltered,’ yparameters of all platinum positions are determined. Intensities of (OkO) reflections can then be calculated, and are in good agreement with observation (Table I). One could not achieve this agreement if the molecular structure of the tetramer were seriously altered by complexing with benzene. TABLE I INTENSITIES OF (OkO) REFLECTIONS k
>
d
1 6
8 10
12 14
Intensity Obsd. Calcd.
S M’ 0
\v
s vs s+
1475 64 1 0
173 1178 1576 722
Intensity Obsd. Calcd.
k
M0 0 MMi M+
16 18 20 22 24 26
75 0 12 325 573 630
3265 TABLE I
Reaction mixture no. Culture filtrate diluted (Isomaltose hydr./ml. culture Two hr. filt., mg. Hydrolysis, % Isomaltose hydr./ml. culture Five hr. filt., mg. Hydrolysis, % ‘ “Fermentable” glucose per ml. cult. filt., mg.
{
i
1 Un-
2 1:l 50%
3 1:3 25%
27.4 63.4
19.4 44.9
14.3 33.1
39.0
35.1
90.2
81.2
29.0 67.1
37.5
33.0
25.7
It will be noted that the undiluted culture filtrate hydrolyzed 90.2% or 39.0 mg. of the isomaltose per ml. of culture filtrate in five hours. The amount of fermentable glucose produced, 37.5 mg., per ml. of culture filtrate checks reasonably well with this figure. (The hydrolysis of isomaltose proceeds more slowly than the hydrolysis of maltose as indicated by the fact that 38.2 mg. of maltose is hydrolyzed per ml. of culture filtrate in one hour a t 30”). I n addition, the sugar formed in a reaction mixture identical to No. 1 in Table I, but incubated for six hours, was isolated as crystalline a-D-glucose monohydrate; [(Y]*~’D $47.8‘ (c, 4 in water); m. p., 82.8’4; yield, 83.4%. Further work on characterization of the enzyme system responsible for the hydrolysis of isomaltose is in progress.
1)EPARTMENT OF CHEMISTRY IOWA STATECOLLEGE
(4) Cf.Nat. Bur. Stand. Circ. No. C440, p. 728. R. E. RUNDLE AMES,IOWA E. JEANNE HOLMAN CORNINDUSTRIES RESEARCH FOUNDATION NEW YORK,N. Y. RECEIVED JULY 25, 1949 NORTHERN REGIONAL RESEARCH LABORATORY BUREAUOF AGRIC.AXD IND.CHEM. HENRYM. TSUCHIYA UNITED STATES DEPT. OF AGRIC.EDNAM. MONTGOMERY HYDROLYSIS OF THE a-l,6-GLUCOSIDICTINKAGE JULIAN CORMAN IN ISOMALTOSE BY CULTURE FILTRATE OF PEORIA,ILLINOIS RECEIVED MAY31, 1949 ASPERGILLUS NIGER NRRL 330
Sir : DERIVATIVES OF DIBORINE The a- 1,B-glucosidic linkage in the amylopectin fraction of starch is resistant to hydrolysis by the Sir : Although tetrachlorodiborine, B2C14, first preaction of the more common amylolytic enzymes. Inasmuch as the linkage between the two glucose pared by Stock, Brandt and Fischer (Ber., 58, residues in isomaltose is of the a-1,6 type, i t was 855 (1925)) by striking an arc across zinc elecof interest to test the ability of a culture filtrate of trodes immersed in liquid boron trichloride, is a Aspergillus niger NRRL 330 to hydrolyze this compound of considerable potential interest, the disaccharide. Isomaltose is now available in original preparative method gave such poor yields crystalline form. The sample of sugar used in (about 1%) of material of such low purity (less these experiments was prepared from starch by than 90%) that detailed study of the compound the method of iMontgomery, Weakley and Hilbert‘ was not feasible. and characterized as 6-(a-D-glucopyranosyl)- a - ~ - Much better results have now been obtained by glucose monohydrate, rotating 120’ in water passing gaseous boron trichloride a t 1 to 2 mm. (anhydrous). This particular strain of A . niger through a glow discharge established between was selected because i t elaborates, when grown in mercury electrodes. The mercurous chloride submerged culture, an enzyme capable of hydro- and other non-volatile products remain in the discharge tube ; the volatile material is passed lyzing maltose in large amounts. T o 20 ml. of 0.06 M isomaltose monohydrate, through a -78.5’ trap, which retains the tetrabuffered a t PH 4.1 (acetate buffer, 0.3 AT), was chlorodiborine and allows the unchanged tri:tdded IO ml. of culture filtrate. The reaction chloride to pass. The latter is repeatedly submixture was held a t 50’. The extent of hydroly- jected to the action of mercury in the discharge sis was measured a t two and five hours by the tube, since the amount converted per pass is increase in reducing power as determined by the small. The yield, however, is approximately method of Somogyi.2 The glucose formed in the 50%. There is evidence that other less volatile reaction mixture after five hours was identified boron-chlorine compounds are obtained. Tetrachlorodiborine is completely hydrolyzed biologically by “fermentation” a t PH 8.8 with an excess of yeast by the procedure of S ~ m o g y i . ~by aqueous sodium hydroxide a t 70’. A collection of some of our experimental data is B2CL 6NaOH +2NaB02 4NaC1 f 2H20 + Hz given in Table I. Determination of the amounts of hydrogen, (1) Edna M. Montgomery, F. B. Weakley and G. E. Hilbert, boron and chlorine in the hydrolysate of an unTHISJOURNAL, 71,1682 (1949). weighed sample by the usual methods gave the (2) M. Somogyi, J . B i d . Chem., 160, 61 (1845) 13) M.Somogyi. ibid , 119, 741 (19371. following results: H, 74.64 cc.; R , 0.0727 g.;
+
+
+
3266
COMMUNICATIONS TO THE EDITOR
Vol. 71
C1, 0.481 g., corresponding to the atomic ratio related to diethylstilbestrol, apparently irrespecH:B:CI = 1.98:2.00:4.03. A 0.0995-g. sample tive of configuration, Schoeller, Inhoffen, Steingave 0.0131 g. of boron and 0.0878 g. of chlorine, ruck and Hoss, U. s. Patent 2,392,864 (Jan. 15, a total of 0.1009 g. The vapor density corre- 1948)]. sponds to a molecular weight of 163 (calcd., 163.5). Since 1941 we have directed efforts toward the The vapor tensions, measured between - 63.5 synthesis of analogs of the non-aromatic steroid and 22.5', are satisfactorily reproduced by the hormones lacking ring C, and wish to report now the synthesis of two such analogs of the androgens : equation: log P ( m m . ) = - 1753/T f 8.057. Liquid tetrachlorodiborine a t room temperature I , comparable to testosterone, and 11, to androfor seventy-two hours undergoes 2lY0 decomposi- stenedione. tion, yielding boron trichloride, a very slightly volatile red substance and a white solid. The latter two must be hitherto unknown boron chlorides of as yet undetermined composition. At 0' the decomFosition is much slower. The following reactions of tetrachlorodiborine have been studied in preliminary fashion. (1) I I1 It yields the hitherto unknown tetrabromodiborine by treatment with boron tribromide, but does not These a,p-unsaturated ketones were synthesized react with boron trifluoride. ( 2 ) With dimethyl using the Robinson-Mannich base method [du or diethyl ether it forms a liquid monoetherate Feu, McQuillin and Robinson, J. Chem. SOC.,53 and a solid dietherate. The latter has an ap(1937) ] with some modifications. Hydrogenation preciable dissociation pressure and is somewhat of 4,4'-dihydroxybiphenyl gave the three possible soluble in the ether. (3) It yields the correperhydro derivatives (m. p.'s 215-216', 176.5sponding tetramethoxy and tetraethoxy deriva177' and 194-195', all m. p.'s cor.). Half-oxidatives on treatment with the appropriate alcohols. of the first two stereoisomers with chromium tion (4) It reacts with lithium borohydride and alumitrioxide afforded the same 4-(4'-hydroxycyclonum borohydride to give boron hydrides among hexyl)-cyclohexanone (111, probably trans), m. p. which diborane and dihydrotetraborane (B4H10) 129-130'. Anal. Calcd. for ClzHzoOz: C, 73.4; have been definitely identified ; pentaboranes H, 10.3. Found: C, 73.6; H, 10.1. When and decaboranes are probably present. It does sufficient diol becomes available, i t will prenot seem to react with lithium aluminum hydride sumably 195' lead to the other possible isomer. (5) It absorbs hydrogen a t room tem- More complete oxidation gave 4-(4'-ketocycloa t 0'. perature, by which reaction all of the chlorine is hexyl)-cyclohexanone (IV), m. p. 115-116'. converted to boron trichloride, and the residue is a Anal. Calcd. for ClzHlsOz: C, 74.2; H, 9.3. compound or mixture of the approximate composi- Found: C, 74.3; H, 9.2. tion BH1.3, (B10H14?). (6) It reacts with ammonia Formylation of the keto alcohol (111) with in complex fashion. These and other reactions, ethyl formate and sodium methoxide, reaction as well as the preparative method, will receive of the derivative with the methiodide of l-diethylfurther intensive study. aminobutanone-3 and cyclization with ca. 4% We gratefully acknowledge the interest and methanolic potassium hydroxide at room temfinancial support of the Office of Naval Research perature gave 6-(4'-hydroxycyclohexy1)-A1-9-octaand the Naval Research Laboratory. lone-2 (I), purified by chromatography on alumina DEPARTMENT OF CHEMISTRY THOMAS WARTIK and recrystallization from cyclohexane-ethyl aceTHEUNIVERSITY OF CHICAGO R.MOORE tate, m. p. 127-127.5'; max. 238.5 mp (Emoiar = H. I. SCHLESIXGER16,870). Anal. Calcd. for CloH2402: C, 77.4; CHICAGO, ILLINOIS RECEIVED JULY 18, 1949 H, 9.7. Found: C, 77.7; H, 9.8. The semicarbazone melted a t 235-236' (dec.). Anal. Calcd. for Cl,H2,N302: C, 66.9; H, 8.9. Found: SYNTHESIS OF PHYSIOLOGICALLY ACTIVE C, 67.0; H, 8.7. ANALOGS OF TESTOSTERONE Similarly, monoformylation of the diketone Sir: (IV), reaction with the Mannich base methiodide The discovery of several hundred compounds and cyclization gave 6- (4'-ketocyclohexyl) possessing the physiological activity of the estro- octalone-2 (11), also obtained in small yield by genic hormones has prompted a number of investi- oxidation of I (as the dibromide), m. p. 88.5gators to search for synthetic analogs having 89.5'. max. 238 mp (Emolar = 16,400). Anal. activity similar to the other steroid hormones. Calcd. for Cl6HZ2O2:C, 78.0; H, 9.0. Found: While some claims of activity for such analogs C, 77.9; H, 8.7. The disemicarbazone decomhave been made, particularly in patents, so far posed a t 256-257'. Anal. Calcd. for Cl~HzsNsOz: as we are aware no such claim has yet been sub- C, 60.0; H, 7.8. Found: C, 59.9; H, 7.5. stantiated [cf., for example, the reported androPreliminary assays in day-old chicks, under the genic activity for certain reduction products direction of Drs. R. K. Meyer and Elva G. Ship-
