Cloning of a Transglutaminase Gene from an Indonesian Streptomyces Strain

Abstract

Microbial Transglutaminase (MTGase, EC 2.3.2.13) is an enzyme that catalyzes the transfer of acyl group. It has tremendous applications in binding meats together in texture improvement process. This research was aimed at cloning MTGase gene from Streptomyces thioluteus TTA 02 SDS 14. A genomic library with the average of 40 kb insert size was initially generated from the total genomic DNA of S.thioluteus TTA 02 SDS 14, and was subsequently screened using an MTGase-targeting primer pair. The genomic library construction was based on the fosmid pCC1FOS TM hosted in E.coli EPI300-TI R . The isolated positive clone was verified by enzymatic digestion with NotI, EcoRV, and BamHI, followed by PCR-amplification of MTGase fragment. The PCR product result of 360 bp was sequenced. The sequence analysis strongly suggests that the positive clone with 40-kb insert harbors an MTGase gene.