By T.K. Ghose ,A. Fiechter, N.Blakebrough
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Extra resources for Advances in Biochemical Engineering, Volume 003
I i I I I ! ~'_~ '" . . . . . . . . . . . . . . • Fig. 20. , 1971) a genetic analysis as loci responsible for the biosynthesis of the antibiotic. The important genes responsible for the formation of tetracyclines may thus belong to different, frequently independent, metabolic pathways. A common feature of these loci is the final effect of their action-provision of intermediates for the biosynthetic process. Van~k et al. e. the transformation of the hypothetical tricyclic nonaketide, 56 z.
Ura Ala; Arg; Cys; Leu; Met/Hcs; Ade; Gua; Gua + Met/Cys Mutations were induced by N-methyl-N'-nitro-N-nitrosoguanidine and by UV light. Both the delayed enrichment method and the total isolation were used for the detection of auxotrophs. b NRRL 2209, wild low-producing strain; 84/25, producing standard strain; the other parents represent different types of blocked mutants. arg-19 ser-3 met-4 H-56 E-14 S-10 Arg; Leu; Gly/Ser/Thr; Met/Thi; Met/Thi/Glu; Met/Cys/Thr/Glu; N ic Arg; Gly/Ser/Thr; Ade/Gua pur-I H-12 none Arg; Gly/Ser; Ura Arg Arg Arg Arg; Ile+Val; Met; Phe none Arg -arg-8 ilv-2 C-22 NL-1 - Arg; lle +Val 84/25 B-96 Arg; Ile; Ile+Val; Arg+Ile+Val; Leu; Lys/Pdx: Met; Thr: Ser/Gly: Guo his-4 R-6 Arg: His; Ile+Val; Leu; Lys; Met; Gly/Ser; Ade; Pdx Arg; Lys Additional requirement Genotype marker Requirement for Parent strain 2nd step auxotrophs 1st step auxotrophs NRRL 2209 Parent prototroph Table 10.
5 1 0 ( O 400 140--- 300 I ~ 450 0 . 5 - - 30 2 3 4 5 6 Compleorange mentary OTC group lag/ml Cosynthesis a Mutants were derived from S. rimosus LS-T 118 producing about 3000 lag OTC/ml. This strain formed a brown pigment under submerged conditions. b Production ( + ) or absence (--) of metabolites was detected by paper c h r o m a t o g r a p h y in system chloroform-nitromethane-pyridin (10:20:3) or butanol-acetic acid-H20 (16:4: 5). "Black" mutants. d "White" mutants. Number of murants Mutant group Table 6.
Advances in Biochemical Engineering, Volume 003 by T.K. Ghose ,A. Fiechter, N.Blakebrough