Microbial Transglutaminase Fermentation And Its Application Of Wound Healing In Rat

Transglutaminase (TGase, protein-glutamineγ-glutamyltransferase, EC. 2.3.2.13) is an enzyme that catalyses the acyl transfer reaction by introducing covalent cross-links between proteins, peptides and various primary amines. The reaction catalyzed by transglutaminase can be used to modify the function of proteins, thus transglutaminase has been widely used in food, cosmetic, pharmacy and other industries. Traditionally, most sources of TGase come from animal organs, but in view of economic value, but by the economics, it is necessary to look for more convenient and economical method for production. Fermentation has the advantage of low cost, which makes it a most promising method in large-scale production of transglutaminase.The influence of vitamin for Streptoverticillium SG 215 was systematically studied with central composite design. Firstly, the prime factors affecting MTG activity were selected by Plackett-Burrman; secondly, the prime factors were optimized by steepest ascent design and Central Composite Design. The results showed that Calcium Pantothenate and Para-amino benzoic acid had great influence on the growth of Streptoverticillium SG 215 and MTG activity, and the optimum fermentation medium were Vitamin B1 20 mg/L,Vitamin B2 0.5 mg/L, Niacin 6 mg/L, Calcium pantothenate 0.028 mg/L, Vitamin B3 0.5 mg/L, Pyridoxal hydrochloride 0.2 mg/L, Folic Acid 0.4 mg/L, Biotin 20 mg/L, Para-amino benzoic acid 0.044 mg/L. The maximum activity of MTG under optimum fermentation medium was 2.42 U/ml which was 41.77% higher than that of the original medium (1.58 U/ml).The fermentation kinetics of glutathione with SG 215 was studied. The kinetic models were obtained. The cell growth model was showed in following equation:(?),the MTG biosynthesis model was showed in followingequation:(?);the glucoseConsumption mode was showed in following equation:(?).The kineticmodel provided a reasonable description for biomass, product and substrate variation along with the fermentation process.Effect of Microbial Transglutaminase from SG215 on cutaneous wound healing in rats was evaluated using excision wound model. The closure time and volume of the skin wounds in the experimental groups significantly decreased when compared to that in the control group (P<0.05).Wound contraction, dry(or wet) weight of granulation tissues and content of protein, Hydroxyproline, Hexuranic, Hexosamines in the wound tissues in the experimental groups were higher than that in the control group (P<0.05).These results clearly substantiate the beneficial effects of microbial transglutaminase in wound healing.