COMMWMCATIONS TO THE EDITOR
3968 NdtO,, aqueous, % ’ NdzOs, al(!OhOl, % ’ Separation factor’
I
I1
53.7 63.2 1.48
46.1 57.0 1.55
Vol. 70
4.68; N, 8.49. Found: C, 69.19; H, 4.79; N, 8.33. Since this heptabenzoylstreptidne is identical with the one4 obtained from streptomycin it follows that in mannosidostreptomycin the mannosidostreptobiosamine is attached to Cq of streptidine, just as streptobiosamine is attached to streptidine in streptomycin. Furthermore, since this product has seven rather than six benzoyl groups, these findings confirm the earlier observation2that mannose is not attached to streptidine.
These are ten-fold better separations than those which Appleton and Selwood obtained with thiocyanates . A three-stage fractionation also was performed. The alcohol volume was ten times that of the aqueous phase. One to two hours, with agitation, were required for extraction; the washing of the yield ROBERT L. PECK out of the alcohol took thirty minutes. Each stage MERCK& Co.. INC. CHARLES E. HOFFHINE, JR. consisted of four extractions, after which all four RESEARCH LABORATORIES PAUL GALE NEWJERSEY KARLFOLKERS yields were combined to form the aqueous solu- RAHWAY, RECEIVED AUGUST 13, 1948 tion of the next stage. Between extractions the aqueous phase was partially evaporated to keep it near saturation. Starting with 50 g. of 50.7% NdzOs (in total oxides), the first stage yielded 27 g.; the second, 16 g.; and the third, 9 g., of ni- THE OCCURRENCE OF MANNOSIDOSTREPTOtrates. The third yield analyzed 69.0% Nd203. MYCINASE Thus the separation factor was 2.14 for three Sir : stages and averaged 1.29 for each stage. During the course of experiments on the produc(7) As defined by Appleton and Selwood. tion of streptomycin B (mannosidostreptomycin) by a number of cultures of Streptomyces griseus, it DEPARTMENT OF CHEMISTRY was observed that certain growing cultures were UNIVERSITY OF MICHIGAN CHARLES C. TEMPLETON ANNARBOR, MICHIGAN able to decompose added streptomycin B as shown OF CHEMISTRY DEPARTMENT by isolation of Streptomycin A from the fermented UNIVERSITY OF 1NISCONSIN medium and by chemical analysis of the isolated MADISON, WISCONSIN JOHN A. PETERSON product (by a method to be published.) Cultures RECEIVED OCTOBER 18,1948 of S. griseus which produce streptomycin A showed the ability to convert streptomycin B to streptomycin A, while this property was lacking STREPTOMYCES ANTIBIOTICS. XXI. LINKAGE OF in cultures of S. griseus which did not produce streptomycin A. This enzymatic activity was MANNOSIDOSTREPTOBIOSAMINETO STREPTIDINE IN MANNOSIDOSTREPTOMYCIN not found in other actinomyces cultures which do not produce streptomycin nor in cultures of PeniSir: Recent evid.ence1I2has shown that in mannosi- cillium chrysogenum and Aspergillus niger. Sevdostreptomycins the mannose (pyranoside) is eral commercial multi-enzyme preparations were linked to Cq of N-methyl-L-glucosamine. How- examined for the presence of the mannosidostrepever, there has been adduced no evidence to show tomycinase, and although two showed some indithe nature of the linkage of mannosidostreptobi- cation of activity, the degree of activity was much less than was observed in cell-free preparaosamine to streptidine. Benzoylation of mannosidostreptomycin using tions from a streptomycin A-producing strain of conditions previously described for preparation of S. griseus. Working with a cell-free preparation, undecabenzoyl~treptomycin~ gave amorphous te- evidence that the enzyme induces hydrolysis of tradecabenzoylmannosidostreptomycin; [a]2 6 ~ the streptomycin B at the point of attachment of the mannose moiety includes the observations + 5 . 5 O (c, 1.8 in chloroform). could be identified in the solution Anal. Ca1c:d. for C ~ ’ I H ~ ~ N ’ I O ~ ~ ( C B H C,~ Cthat O ) I ~mannose : as the phenylhydrazone and that the maltol assay 68.20; HI 4.81; N, 4.46. Found: C, 68.17; H, of the solution did not change while the antibiotic 4.72; N, 4.93. activity increased by a t least 70% of the amount Cleavage of tetradecabenzoylmannosidostrepto- expected. The p H optimum appears to be bemycin in chloroform solution with hydrogen bro- tween 7.5 and 8. Reducing conditions inhibit mide yielded a heptabenzoylstreptidine; m. p. hydrolysis as does the presence of Cu and Hg 259-260O; [a]””.+55’ (c, 1.0in chloroform). ions. As indicated by increase in biological Anal. Calcd. for C ~ ’ I H & ~ ~ IC, I : 69.15; H, activity, active cell-free preparations also appear to induce hydrolysis of dihydrostreptomycin B. (1) Fried and Stavely, Abstracts of Papers, 113th Meeting of the American Chemical Soaety, Chicago, Ill., April 20,1948, page 26C. (4) Fried and Stavely, THIO JOURNAL, 68, 1548 (1947). (8) Fried and Titus, J . B i d . Chum., 168,391 (1947). (4) Peck, Kuehl, Hoffhine, Peel and Folkers, THISJOURNAL, 70, at21 (1948).
DIVISION OF MICROBIOLOGICAL DEVELOPMENT E. R.SQUIBB AND SONS D. PERLMAN l l i ~ wBRUNSWICK, N. J. A. F. LANGLYKKE RECEIVED OCTOBER 20, 1948
