Improved Yield of High Molecular Weight Hyaluronic Acid Production in a Stable Strain of Streptococcus zooepidemicus via the Elimination of the Hyaluronidase-Encoding Gene

Molecular Biotechnology
Navid Pourzardosht, Mohammad Javad Rasaee

Abstract

Despite the significant potential of Streptococcus zooepidemicus for hyaluronic acid (HA) production with high molecular weight (MW), the HA degrading properties of hyaluronidase prevents the bacteria to achieve enhanced HA yield with high MW. In the present study, we aim to knockout the hyaluronidase enzyme and assess its effects on the yield and MW of the produced HA. The kanamycin resistance gene between the left and right arms of hyaluronidase gene was inserted into pUC18 plasmid to construct pUC18-L-kanar-R as a recombinant suicide plasmid. The construct was then transferred into S. zooepidemicus to induce the homologous recombination between the hyaluronidase gene and the kanamycin resistance gene. Gene deletion was confirmed by PCR and enzyme assay. The product was cultured on selectable medium in which the MW of HA was increased from 1.5 to 3.8 MDa. The yield of HA production using the mutant strain was higher in all different concentrations of glucose from 40 to 120 g/l. Moreover, glucose increase results in higher HA production within both wild-type and recombinant strains. However, the growth rate of HA concentration (the slope of the plot), as a consequence of increased glucose concentration, is always higher for th...Continue Reading

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Citations

Jan 18, 2018·The Journal of Biological Chemistry·Lisa A AndersonKristala L J Prather
Jul 14, 2019·Metabolic Engineering·Fangyu ChengGregory Stephanopoulos
Apr 30, 2021·Carbohydrate Polymers·Zhi-Yuan YaoJin-Song Shi
Jun 21, 2019·International Journal of Biological Macromolecules·Ehsan DehnaviKhosro Khajeh

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