本論文探討以Streptococcus zooepidemicus ATCC 39920醱酵生產玻尿酸之程序中，攪拌速率對菌體生長以及玻尿酸生產之影響。研究首先發現，在相同攪拌速率下，溶氧值保持在飽和溶氧值的25%以上時，即不會影響玻尿酸的生產。至於攪拌速率對玻尿酸生產的影響可分為兩部分來說明：(一) 當攪拌速率從200 rpm增加到600 rpm時，玻尿酸的產量隨著攪拌速率的提高而增加，然而氧氣傳送係數與細胞攝取養分的速率並不影響玻尿酸生產；顯然地，玻尿酸產量之提高是因為細胞受到剪應力刺激所致。(二) 但是，當攪拌速率從600 rpm增加至800 rpm時，玻尿酸的產量反而下降；推測其原因乃在於剪應力過大，反而不利於玻尿酸之生產。此外，當攪拌速率在200至800 rpm這範圍內，所得到之玻尿酸分子量介於1.21-1.34×106 Da之間，此數據顯示，攪拌速率對玻尿酸的分子量並無顯著之影響。綜合以上結果，本研究認為攪拌產生之剪應力是鏈球菌醱酵生產玻尿酸製程放大的關鍵因素。

The effect of agitation rate on the production of hyaluronic acid (HA) by Streptococcus zooepidemicus ATCC 39920 was investigated in this study. The result first showed that under a specified agitation rate, the production of HA was not influenced by the level of dissolved oxygen (DO) when DO was kept above 25% of saturation. The influence of agitation rate on HA production could be described as follows: (1) The yield of HA increased with agitation rate in the range from 200 to 600 rpm. Since the results showed that both oxygen transfer coefficient and nutrient assimilation rate did not affect the production of HA, the increase in the yield of HA could result from the increase in shear force. (2) The yield of HA decreased when agitation rate increased from 600 to 800 rpm, which might attribute to the inhibition in HA production by extremely high shear force. In addition, the molecular weight of the obtained HA was between 1.21×106 Da and 1.34×106 Da in the range of 200 to 600 rpm. The result indicates that the molecular weight of HA is not significantly affected by agitation rate. Based on the above findings, agitation shear force is thought to be the critical factor for scale up of HA fermentation.

Armstrong, D.C. and Johns, M.R., “Effect of culture conditions on molecular weight of hyaluronic acid produced by Streptococcus zooepidemicus,” Appl. Environ. Microbiol., 63: 2759-2764 (1997).
Bailey, J.E. and Ollis, O.F., Biochemical Engineering Fundamentals, 2nd Ed., McGraw-Hill, New York, Transport Phenomena in Bioprocess System., Pp. 459-460 (1986).
Balazs, E., “The physical properties of synovial fluid and the specific role of hyaluronic acid,” Curr. Ther. Res., 47: 437–443 (1990).
Bitter, T. and Muir, H.M., “A modified uronic acid carbazole reaction,” Anal Biochem, 4: 330-334 (1962).
Bothner, H. and Wik, O., “Rheology of hyaluronate,” Acta Otolaryngol., 44: 25-30 (1987).
Blank, L.M., McLaughlin, R.L., Nielsen, L.K., “Stable production of hyaluronic Acid in Streptococcus zooepidemicus chemostats operated at high dilution rate,” Biotechnol. and Bioeng., 90: 685-693 (2004).
Drobink, J., “Hyaluronan in drug delivery,” Adv. Drug Delivery Rev., 7: 295–308 (1991).
Fong Chong, B. and Nielsen, L.K., “Aerobic cultivation of Streptococcus zooepidemicus and the role of NADH oxidase,” Biochem. Eng. J., 16: 153–162 (2003).
Fong Chong, B., Blank, L.M., Mclaughlin, R. and Nielsen, L.K., “Microbial hyaluronic acid production,” Appl. Environ. Microbiol., 66: 341–351 (2005).
Fouissac, E., Milas, M., and Rinaudo, M., “Shear rate, concentration, molecule weight and temperature viscosity dependencies of hyaluronate, a wormlike polyelectrolyte,” Macromolecules, 26: 6945-6951 (1993).
Harry, M., James, G., and Michael, R., “Hyaluronic acid capsule and the role of Streptococcal entry into Keratinocytes in invasive skin infertion,” J. Clin. Invest., 98: 1954-1958 (1996).
Hascall, V.C. and Laurent, T.C., “Hyaluronan : structure and physical properties,”http://www.glycoforum.gr.jp/science/hyaluronan/HA01/HA01E.html, 1997.
Johns, M.R., Goh, L.T., and Oeggerli, A., “Effect of pH, agitation and aeration on hyaluronic acid produced by Streptococcus zooepidemicus,” Biotechnol. Lett., 16: 507-512 (1994).
Kendall, F.E., Heidelberger, M., and Dawson, M.H., “A serologically inactive polysaccharide elaborated by mucoid stains of group A hemolytic streptococcus,”J. Biol. Chem.,118: 61-69 (1937).
Kim, J. H., Yoo, S.T., Kweon, Y.G., Oh, D.K., Park, D.W., Lee, C.H., and Gil, G.H., “Selection of a Streptococcus equi mutant and optimization of culture conditions for the production of high molecular weight hyaluronic acid,” Enzyme and Microb. Technol., 19: 440-445 (1996).
Laurent, T.C. and Fraser, J.R., “Hyaluronan,” FASEB J., 6: 2397–2404 (1992).
Laurent, T.C., “Biochemistry of hyaluronan,” Acta Otolaryngol., 442: 7-24 (1987).
Laurent, T.C., Ryan, M., and Pietruszkiewicz, A., “Fraction of hyaluronic acid. The polydispersity of hyaluronic acid from the bovine vitreous body,” Biochim. Biophys., 42: 476-485 (1960).
Lowry, K.M. and Beavers, E.M., “Thermal stability of sodium hyaluronate in aqueous solution,” J. Biomed. Mater. Res., 28: 1239-1244 (1994).
Meyer, K. and Palmer, J.W., “The polysaccharide of the vitreous humor,” J. Biol. Chem., 107: 629-634 (1934).
Meyer, K., “The biological significance of hyaluronic acid and hyaluronidase,” Physiol. Rev., 27: 335-359 (1947).
Nimrod, A., Greenman, B., Kanner, D., and Landsberg, M., “High molecular weight sodium hyaluronate,” U.S. Patent. 4784990 (1988).
Nimrod, A., Greenman, B., Kanner, D., and Landsberg, M., “Method of producing high molecular weight sodium hyaluronate by fermentation of Streptococcus,” U.S. Patent. 4780414 (1988).
Rapport, M.M., Weissman, B., Linker, A., and Meyer, K., “Isolation of a crystalline disaccharide, hyalobiuronic acid, from hyaluronic acid,” Nature, 168: 996-997 (1951).
Scott, J.E., Heatley, F., and Hull, W., “Secondary structure of hyaluronate in solution,” Biochem. J., 220: 197-205 (1984).
Scott, J.E., “Secondary and tertiary structures of hyaluronan in aqueous solution,” http://www.glycoforum.gr.jp/science/hyaluronan/HA02/HA 02E.html, 1998.
Tribe, L.A., Briens, C.L., and Margaritis, A.A., “Determination of the volumetric mass transfer coefficient using the dynamic "Gas Out-Gas In" method: analysis of errors caused by dissolved oxygen probes,” Biotechnol. and Bioeng., 46: 388-392 (1995).
Woolcock, J.B., “The capsule of Streptococcus equi.,” J. Gen. Microbiol., 85: 372-375 (1974).
Yamamoto, H., “Cosmetic effects of dietary hyaluronic acid (extracellular matrix extract),” New food ind., 40: 33-41 (1998).
王雲萍，生物科技的寵兒—玻尿酸，化工資訊，5: 44-49 (1994)。
陳士軒，透明質酸之製備與傷口癒合應用，陽明大學碩士論文 (1996)。
凌沛學，賀豔麗，郭學平，侯麗君，楊曉虹和張天民，透明質酸，中國輕工業出版社 (2000)。
黃定國，透明質酸之開發與應用，菌種保存及研究簡訊 3:1-9 (2001)。