Separation and Purification of Hyaluronic Acid from Fermentation Broth
The objective of this research was to compare different methods for the separation and purification of hyaluronic acid from Streptococcus zooepidemicus ATCC39920 for their ease of use and reliability. The most appropriate conditions of pretreatment stage
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Separation and Purification of Hyaluronic Acid from Fermentation Broth Yunshuang Wang, Jian Zhang and Hao Liu
Abstract The objective of this research was to compare different methods for the separation and purification of hyaluronic acid from Streptococcus zooepidemicus ATCC39920 for their ease of use and reliability. The most appropriate conditions of pretreatment stage were heated to 70 °C for 1 h and kept at room temperature for 5 h. The fermented broth was diluted with the equal volume deionized water. Ten milligrams chitosan was slowly added to 1 L of solution with stirring for 30 min, followed by the addition 20 g diatomaceous earth-type filter aid. The mixture was stirring for 1 h. Then, the mixture was filtrated with diatomite, microfiltration (MF) membranes (5 lm in pore diameter), and ultrafiltration (UF) membranes (MWCO 50KD) to achieve the separation stage.The retentate was washed by the equal volume deionized water and when the volume concentration ratio (VCR) reached 4, the UF stage ended. Finally, the purification procedure adopted Sevag process to remove protein. The overall yield of HA could reach 62.53 % and the removal rate of protein could reach 95.22 %. Keywords Hyaluronic acid Separation process
Fermentation broth
Pretreatment process
162.1 Introduction Hyaluronic acid (HA), is a high-molecular weight linear polysaccharide, composed of D-glucuronic acid and N-acetylglucosamine linked alternately by b-(1-3)and b-(1-4)-glycosidic bonds. HA is typically found in the connective tissues of animals as well as in the capsules of streptococcal bacteria. The highest content of Y. Wang J. Zhang H. Liu (&) Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, People’s Republic of China e-mail: [email protected]
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 251, DOI: 10.1007/978-3-642-37925-3_162, Ó Springer-Verlag Berlin Heidelberg 2014
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HA is found in rooster combs. Its molecular mass in human normal synovial fluid has been estimated to be 6–7 9 106 and in rheumatoid fluid 3–5 9 106 Dalton [1]. Traditionally, HA has been extracted from bovine eyes and rooster combs [2]. However due to limited tissue sources, risks of viral infection and high cost, HA production from microbial sources through the fermentation process has received increased attention especially when using the gram-positive bacterium Streptococcus zooepidemicus [3–7]. The separation and purification of HA involves the precipitation of HA from fermentation broth by repeatedly using large amounts of organic solvents such as ethanol, acetone, isopropanol, etc. [8–12]. However, the process is complicated and time-consuming, which leads to high cost. In order to improve the efficiency of the purification of HA a better understanding of each steps in the purification process is needed. The purpose of this study was to comp
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