Nov. 30, 1950 Dear Joshua - - Re tested liquid medium : I regret that we have no personal experience since the sort of problem you're interested in has never arisen. However, Seymour Hutner (who has chided me for the inadequacy of our synthetic media) recommends (Proc. Am. Phil. Soc. 94, 152, 1950) : Ethylemediamine tetraacetic acid 0.05g. K2HPo4 0.02 g. 14g So4 7H2O 0.08g. NH4Cl (NH4NO3 preferable, I think) 0.02 g. 2n 5.0 mg. Fc 1.0 mg. Ca 2.0 mg. Mo 0.6 mg. B 2.0 mg. Cu 0.4 mg. Mn 1.4 mg Co 0.4 mg I would recommend a [ . . . ] ph : 7.5, if precipitation is not a problem To pH 6.6 - 6.9 with KOH : dist. H20 to 100 ml. He says this gives abundant growth, with a suitable C-Source, of many strains that grow badly in the sort of crude media we use. No chance of a paper on UV / adaptation at the moment, as I am up to my ears in tryptophane metabolism with Hayaishi, who leaves for Kornberg's lab in January. Also, I want to see what Cyiese[?] has in prog on UV / adaptation in yeast : he may have anticipated most of our points. Shirley[?] Gunter[?] has been using it to study the sequence of adaptation in metabolic pathways, with dull as expected results ; the intermediates are adapted to at much the same rate as the primary methods. All this shows is that v. small amounts of subtrate are needed to start induction. [END PAGE ONE] [BEGIN PAGE TWO] The hyptophon story is fascinating. In pseuderminades[?], there are 2 radically distinct pathways, diverging at Kynmere [TABLE="Aromatic pathway"] [TABLE="Quinotive pathway."] probable from Japanese work, but not yet proven by us Some stigs[?] on the aromatic pathway have a block below anthramidie, which accumulates in yields of +- 80%. Despite this very [ . . . ] block, they are fully adapted to catrechol[?], the small kak thru the block being sufficient to give complete activation of the later adaptive systems. In 25 tested strs the pathway is either/ or ; no "mixed" races. But we have one extraordinary strain which accumulates in quantitative yield from tryptophan a mixture (ration variable) of kynmeric[?] and authranilie[?] acids, there being an apparent absolute double block. There is also one other aberrant stain with an early absolute block that accumulates an indole derivative, probably g-OH indole, and is thus fairly close to a reversal of the biosynthetic pathway. So much for Kees' idea that degradation pathways should be the reverse of synthetic ones. We have many enzymes cell- free kynmteninase[?] (a pryiodoxal - P enzyme that cleaves to alanine and antiramilie), the synthin oxidizing trypt -> kymnermine[?], or the catriehol -oxidizing system. Mcilwain's technique of alumina grinding has proved a godsend in making preparations of the enzymes, since some [END PAGE TWO] [BEGIN PAGE THREE] notably hyptophan oxidose, are completely wreaked by sloppy drying et al. Alumina guiding is quick, easy and miraculous - - see description in J. Gen. Microbiol. 1948. Did I write to you about the possibility of a visit to Madison in Feb? I'd like to come after the program committee meeting of the SAB in Iowa City. It would help if you could dig up a small honorarium for a seminar, but this won't be vital unless I'm broker than I expect to be by then Best wishes Roger