Effect of Various Physical Parameters and Statistical Medium Optimization on Production of Hyaluronic Acid Using S. Equi Subsp. Zooepidemicus ATCC 39920

V J Aroskar, S D Kamat, D V Kamat


It has been shown that initial conditions for bacterial cultivation are extremely important for the successful production of hyaluronic acid (HA) by fermentation. We investigated several physical parameters that affect productivity of HA under shake flask. i.e. transfer criteria of seed, agitation and aeration of fermentation flasks. Among the various physical parameters studied, inoculum age of 8-10 h, pH 6.4, optical density (600 nm) 2.0 and 3% level inoculum transfer found to be optimum. After inoculating with Streptococcus equi subsp. zooepidemicus ATCC 39920, the temperature 37 0C and 90 rpm found optimum during growth as well as for the HA production. The fractional factorial design of six factors with two levels showed yeast extract, potassium dihydrogen phosphate and sodium bicarbonate as significant model terms. The factor potassium dihydrogen phosphate was relatively more significant than yeast extract.



Streptococcus zooepidemicus, Hyaluronic acid, shake flask fermentation, full factorial design

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Akaska, H., Komasaki, H., and Avali, T. (1989). Fermentation method for producing HA . U. S. Patent No. 4,801,539.

Armstrong, D.C., and Johns, M.R. (1997). Culture conditions affect the molecular weight properties of HA produced by Streptococcus zooepidemicus. Appl. Microbiol. Biotechnol. 63(7): 2759–2764.

Bitter, T. and H. M. Muir. (1962). A modified uronic acid carbazole reaction. Anal. Biochem. 4:330-333

Chen, S.J., Chen, J.L., Huang, W.C., and Chen, H.L. (2009). Fermentation process development for HA production by Streptococcus zooepidemicus ATCC 39920. Korean J. Chem. Eng. 26(2): 428-432.

Chong, B.F., Blank, L.M., Mclaughlin, R., and Nielsen, L.K. (2005). Microbial HA production. Appl. Microbiol. Biotechnol. 66: 341-351.

Duan, Jie, X., Yang, Li., Zhang, Xu., and Tan.W.S. (2008). Effect of oxygen and shear stress on molecular weight of HA Produced by Streptococcus zooepidemicus. J. Microbiol. Biotechnol. 18(4): 718–724

Hiroshi M., and Masahiro F. (1991). Process for preparing HA . U.S. Patent No. 5,071,751.

Huang, W.C., Chen, S.J., and Chen, T.L. (2007). Modeling the microbial production of HA . J. Chinese Institute of Chemical Engineers. 38: 355–359.

Im, J.H., Song, J.M., Kang, J.H., and Kang, D.K. (2009). Optimization of medium components for high-molecular-weight HA production by Streptococcus sp. ID9102 via a statistical approach. J. Indus. Microbiol. Biotechnol. 36(11): 1337-1344.

Jagannath, S., and Ramachandran, K.B. (2010). Influence of competing metabolic processes on the molecular weight of HA synthesized by Streptococcus zooepidemicus. Biochem. Eng. J. 48: 148–158.

Kim, S.J., Park, S.Y., and Kim, C.W. (2006). A novel approach to the production of HA by Streptococcus zooepidemicus. J.

Microbiol. Biotechnol. 16(12): 1849-1855.

Liu, L., Du, G., Chen, J., Wang, M., and Sun, J. (2008A). Influence of modes on microbial production of HA by Streptococcus zooepidemicus. Biotechnol. Bioprocess eng. 13: 269-273.

Liu, C., Sun, Z., Du, J., and Wang, J. (2008B). Response surface optimization of fermentation conditions for producing xylanase by Aspergillus niger SL-05. J. Ind.Microbiol. Biotechnol. 35(7): 703-711.

Liu, L., Du, G., Chen, J., Wang, M., and Sun, J. (2008C). Enhanced HA production by two stage culture strategy based on the modeling of batch and fed batch cultivation of Streptococcus zooepidemicus. Biores. Technol. 99: 8532-8536.

Markovitz A.J.;Cifonello J.A. and Dorfman A,(1959) The biosynthesis of Hyaluronic acid by group A Streptococcus. Journal of biological chemistry. (234) 2343-2350.

Nimrod, A., Greenman, B., Kanner, D., and Landsberg, M. (1988). Ultrapure HA and the use thereof. US Patent No. 4141973.

O,Regan M, Martini I, Crescenzi F, De Luca C, Lansing M (1994). Molecular mechanism and genetics of hyaluronan biosynthesis. Int J Biol Macromol. 16: 283-286.

Kanchankumar P Patil,kirtee k Kamalja and Bhushan L Chaudhari (2011).Optimization of medium components for Hyaluronic acid production by Streptococcus zooepidemicus MTCC 3523 using a statistical approach.Carbohydrate polymers. 86:1573-1577.

Poli, S., Bocchiola, G., Casareto, E., Leoni, M., Mazni, A., and Ronzio, E. (1996). Process for preparation of HA by fermentation with Streptococcus. European Patent No. 0694616A2.

Rangaswamy, V., and Jain, D. (2008). An efficient process for production and purification of HA from Streptococcus equi subsp. zooepidemicus. Biotechnol. Lett. 30: 493–496.

Rangaswamy, V., and Jain, D. (2008B). Optimisation of culture conditions for production and process of purification of high molecular weight HA . United States Patent Application 20080138865.

Stahl, S. (2003). Methods and means for production of HA . US Patent No. 6537795.

Rangaswamy, V., and Jain, D. (2008A). Efficient process for purification of high molecular weight HA . Patent No. WO 2008/035372.

Stoolmiller A. C., and Dorfman A. (1969). The biosynthesis of HA by Streptococcus. J. Biol. Chem. 244: 236-346.

Swann D.A.and J.W.Kuo 1991.Hyaluronic acid p 286-305,in D.Byrom eds.,Biomaterials-novel materials from biological sources,stokton press,New york,N.Y.

Swann, D.A., Sullivan, B.P., Jamieson, G., Richardson, K., and Singh, T. (1990). Biosynthesis of Hyaluronic acid. US Patent No.: 4897349.

Trevelyan WE;Forrest RS and Harrison JS(1952)Determination of yeast carbohydrates with the anthrone reagent. Nature. 170:626-627.

Trinder P. (1969) Determination of blood glucose using an oxidase peroxidase system with a noncarcinogenic chromogen. Ann Clin Biochem. 6:24-30

Vazquez, J.A., Montemayor, M.I., Fraguas, J., Murado, M.A. (2009). High production of hyaluronic and lactic acids by Streptococcus zooepidemicus in fed-batch culture using commercial and marine peptones from fishing by-products. Biochem. Engg. J. 24(2-3): 125-130.

DOI: https://doi.org/10.5195/iioablett.2012.17


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