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42 PROCEEDINGS OF INTERNATIONAL SCIENTIFIC CONFERENCE ON APPLIED BIOTECHNOLOGYbased on the modular organization of these genes [7,8]. Recently multifunctional DH domain has been reported from ambruticin biosynthesis gene cluster, having the function of dehydratase, epimerase and enoyl isomerase leading to the formation of THP ring [9].Figure 1. Reduction of last elongated chain in module 8 of herboxidiene PKS by KR and DH domain followed by chain termination from TE domain. The proposed mechanism of formation of THP ring between 3-4 enoyl functionThus, due to subtle curiosity about the mechanism of THP ring formation (Fig. 1) in herboxidiene, one of the factors responsible for formation of the ring, dehydratase domain of module 8 which forms double bond at 2-3 position of herboxidiene was made dysfunctional by changing the amino acid at the active site. Herein, we focused on engineering the 8th module of herboxidiene PKS for modification in the ring structure. The expected modifications into the final compound from the loss of function mutations of last module can be seen in the (Fig. 2).