Characterization and large-scale production of recombinant Streptoverticillium platensis transglutaminase

Abstract

Recombinant Streptomyces platensis transglutaminase (MtgA) produced by the Streptomyces lividans transformant 25-2 was purified by ammonium sulfate fractionation, followed by CM-Sepharose CL-6B fast flow, and blue-Sepharose fast flow chromatography. The purification factor was approximately 33.2-fold, and the yield was 65%. The molecular weight of the purified recombinant MtgA was 40.0 KDa as estimated by SDS-PAGE. The optimal pH and the temperature for the enzyme activity were 6.0 and 55 degrees C, respectively, and the enzyme was stable at pH 5.0-6.0 and at temperature 45-55 degrees C. Enzyme activity was not affected by Ca(2+), Li(+), Mn(2+), Na(+), Fe(3+), K(+), Mg(2+), Al(3+), Ba(2+), Co(2+), EDTA, or IAA but was inhibited by Fe(2+), Pb(2+), Zn(2+), Cu(2+), Hg(2+), PCMB, NEM, and PMSF. Optimization of the fermentation medium resulted in a twofold increase of recombinant MtgA activity in both flasks (5.78 U/ml) and 5-l fermenters (5.39 U/ml). Large-scale productions of the recombinant MtgA in a 30-l air-lift fermenter and a 250-l stirred-tank fermenter were fulfilled with maximal activities of 5.36 and 2.54 U/ml, respectively.

Improving the active expression of transglutaminase in Streptomyces lividans by promoter engineering and codon optimization

“…To further increase the production level of recombinant TGase at a large scale, the optimization medium and culture conditions will be performed in fermentors [29, 30]. …”

Creation of endoglucanase-secreting Streptomyces lividans for enzyme production using cellulose as the carbon source

“…Secretion of human interleukin-1β and Mycobacterium tuberculosis glycoprotein using S. lividans as a host has also been reported (Lichenstein et al 1988;Lara et al 2004); however, production of useful proteins from cellulose using EG-secreting S. lividans has not been reported. In many cases, a large amount of sugars must be added to the culture medium to serve as carbon sources in order to obtain successful protein production by S. lividans (Ogino et al 2004;Lin et al 2008). In contrast, our multiple enzyme secretion system can be applied to the production of various useful compounds from cellulose or cellulosic biomass, resulting in more cost-effective bioprocessing.…”

Microbial Transglutaminase Production: Understanding the Mechanism

“…Characterization and large-scale production of recombinant Streptoverticillium platensis transglutaminase n/a 5.32 (Lin, et al, 2008) a n/a = not applicable.…”

“…An exception however is if Streptomcyes is used as the host. Recently, production of recombinant Streptoverticillium platensis transglutaminase has reached 5.32 U/ mL: for the first time a production of recombinant transglutaminase has been reported comparable with the wild strain (Lin, et al, 2008).…”