This work studied the production of Transglutaminase (TGase) using wheat bran as carbon source. The medium components and culture conditions were optimized by statistical Box-Behnken response surface methodology. The release of active Transglutaminase was enhanced by adding (i) protease to remove pro-region to make inactive enzyme to active form, (ii) Cetyl trimethyl ammonium bromide (CTAB) which facilitated more secretion. Under finally optimized conditions viz. 5 g wheat bran, protease: 39.14 U, magnesium chloride (MgCl