HYALURONIC ACID PRODUCTION BY CORYNEBACTERIUM GLUTAMICUM

Background and Aim:Hyaluronic acid(HA), is a glycosaminoglycan composed of repetitive disaccharide units of D-glucuronic acid and N-acetyl glucosamine, polymerized by hyaluronan synthase(HAS). HA has been shown to be an important biopolymer according to its unique properties such as high hygroscopicity, biocompatibility and non-immunogenicity that made it applicable to widespread use in medicine, pharmaceuticals and cosmetics. Due to the pathogenicity of the bacteria which make hyaluronic acids for capsule formation, microbial production of HA is tending to development of engineered bacteria which are generally recognized as safe(GRAS).Methods:hasA(HAS coding gene) was optimized and synthesized, followed by subcloning in pEKEx2 vector which is an expression shuttle vector for E. coli/ C. glutamicum. C. glutamicum was transformed by pEKEx2-hasA by electroporation. After the extraction of produced HA, production of developed strain was examined by CTAB turbidity method.Results:The result of double digestion of pEKEx2-hasA, showed a 1284 bp band on gel electrophoresis. colony PCR on C. glutamicum by designed primers, showed 750 bp band, amplified a region located at the middle of the gene, as expected. Comparison of the results of CTAB turbidity method, between the developed strain and the wild type, showed the presence of hyaluronic acid in extracted media.Conclusion:In the present study, C. glutamicum ability for HA production has been investigated. The results showed a successful production of hyaluronic acid after the expression of hyaluronan synthase. Recording to C. glutamicum advantages such as fast growth, inexpensive medium and being lack of hyaluronidase, C. glutamicum can be a suitable choice for this purpose.

کلیدواژه ها:

hyaluronic acid ، hyaluronan synthase ، corynebacterium glutamicum