本論文為探討以半連續式(Fill-and-Draw)醱酵方式培養Streptococcus zooepidemicus ATCC 39920生產玻尿酸。研究發現半連續式操作中培養基組成為不可或缺的；當培養基組成缺乏必需胺基酸成分時，將導致半連續式實驗無法連續地操作。另外，從實驗結果顯示半連續式實驗可連續地操作，其選擇培養基置換之時機是重要的，吾人建議置換時機應選擇在對數生長期末期。

Hyaluronic acid production by Streptococcus zooepidemicus ATCC 39920 was performed in a batch fill-and-draw culture. Medium composition was prerequisite for performing fill-and-draw. The deficiency in essential amino acid would result in unrepeatable operation. The timing of medium replacing was important for performing fill-and-draw operation repeatedly. It was suggested that the medium should be replaced at the late log phase of cell growth.

Armstrong, D. C., Cooney, M. J. and Johns, M. R., ”Growth and amino acid requirements of hyaluronic-acid-producing Streptococcus zooepidemicus,“ Appl. Microbiol. Biotechnol., 47: 309~312 (1997)

Bothner, L. M. and Wilk, O., ”Rheology of hyaluronate,” Acta Otolaryn- gol., 44: 25~30 (1987)

Bentley, J. P., ”Mucopolysaccharide synthesis in healing wounds.” In: Dunphy, J.E., Van Winkle, W. Jr., eds. Repair and regeneration. New York: McGraw-Hill: 151~160 (1968)

Bitter, T. and Muir, H. M., ”A modified uronic acid carbazole reaction,” Anal Biochem, 4: 330~334 (1962)

Cooney, M. J., Goh, L. T., Lee, P. L. and Johns, M. R., ”Structure model-based analysis and control of the hyaluronic acid fermentation by streptococcus zooepidemicus: physiological implications of glucose and complex-nitrogen-limited growth,” Biotechnol., 15: 898~910 (1999)

Denki, K. K., ”Manufacture of hyaluronic acid with Streptococcus equi,” Japanese patent, 924: 93~195 (1993)

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)

Hascall, V. C. and Laurent, T. C., ”Hyaluronan: structure and physical properties,” http://www.glycoforum.gr.jp/science/hyalronan/HA01/HA0 1E.html (1997)

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)

Laurent, T. C., “Biochemistry of hyaluronan,” Acta Otolaryngol., 42: 7~24 (1987)

Laurent, T. C. and Fraser, JRE., ”Hyaluronan,” FASEB J, 6: 2397~2404 (1992)

Meyer, K. and Palmer, J. W., ”The polysaccharide of the vitreous humor,” J. Biol. Chem , 107: 629~634 (1934)

Ogston, A. G., Preston, B. N. and Wells, J. D., ”On the transport of compact particles through solutions of chain-polymers,” Proc. Roy. Soc., 333: 297~309 (1973)

O’Regan, M., Martini, I., Crescenzi, F., De Luca, L. and Lansing, M., ” Molecular mechanisms and genetics of hyaluronan biosynthesis,” Int J Biol Macromol, 16: 283~286 (1994)

Rapport, M. M., Weissman, B. and Linker, A., “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)

van de Rijn, I., ”Streptococcal hyaluronic acid: Proposed mechanisms of degradation and loss of synthesis during stationary phase,” Journal of Bacteriology, 156: 1059~1065 (1983)

Weigel, P. H., Forst, S. J. and LeBoeuf, R. D., ”The specific interaction between fibrin(ogen) and hyaluronan: possible consequences in haemostasis, inflammation and wound healing. In: Evered, D., Whelan, J. eds. The biology of hyaluronan, Ciba Foundation Symposium 143. Chichester: John Wiley & Sons: 248~261 (1989)

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)

三谷, 恗田薰, 須貝哲郎, ”玻尿酸之分子量的保濕能,” 香妝會志, 12: 50~59 (1988)

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)

王雲萍, ”生物科技的寵兒─玻尿酸”. 化工資訊, 5: 44~49 (1994)