Deciphering operation of tryptophan-independent pathway in high indole-3-acetic acid (IAA) producing Micrococcus aloeverae DCB-20.

Genus Micrococcus is considered a high IAA producer. However, interestingly, there is no report on the tryptophan- independent pathway operation in this genus. Consequently, the present study was undertaken to evaluate high IAA production by Micrococcus aloeverae DCB-20 and generate reasonable evidence for the occurrence of the tryptophan-independent pathway. Strain DCB-20 produced a high quantity of 880.51 µM or 154.3 µg/mL IAA in LB broth supplemented with L-tryptophan. The tryptophan-independent pathway operation was supported by IAA production in Tris-minimal broth (TM broth) medium supplemented with acid hydrolyzed casein hydrolysate (casein acid hydolysate), which lacks tryptophan. The HPLC analysis showed the absence of tryptophan either from exogenous or endogenous sources in TM broth in the presence of casein acid hydrolysate inoculated with M. aloeverae DCB-20. The absence of tryptophan was further confirmed by the appearance of non-pigmented colonies of Chromobacterium violaceum strain TRFM-24 on Tris-minimal agar (TM agar) containing acid-hydrolyzed casein. This is probably the first report on IAA biosynthesis by M. aloeverae DCB-20 employing tryptophan-independent pathway. This simple technique can also be adapted to detect operation of the tryptophan-independent pathway in other bacteria.