Abstract
Microbial hyaluronic acid (HA) production has been preferred rather than extraction from animal tissue for medical and cosmetic applications. In this context, to obtain an economically competitive HA production by Streptococcus zooepidemicus, culture conditions were studied to improve the polymer production in sugarcane molasses. The highest HA production by S. zooepidemicus ATCC 39920 achieved was 2.825 g. L−1 in a 4.5 L bioreactor with controlled pH (8.0) and medium containing molasses (85.35 g.L−1 total sugar) pretreated with activated charcoal and yeast extract (50 g.L−1). The HA produced exhibited a high molecular weight of 1.35 × 103 kDa and the DPPH radical scavenging activity of the polymer at 1 g.L−1 was 41 %. The FTIR and UV-Vis spectra showed no substantial differences in the spectral pattern between produced and standard HA. This study is a promising strategy for sugarcane molasses application by producing high value-added products such as hyaluronic acid.
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Chong, B. F., Blank, L. M., Mclaughlin, R., & Nielsen, L. K. (2005). Microbial hyaluronic acid production. Applied Microbiology and Biotechnology, 66(4), 341–351.
Choi, J., Kim, J.-K., Kim, J.-H., Kweon, D.-K., & Lee, J.-W. (2010). Degradation of hyaluronic acid powder by electron beam irradiation, gamma ray irradiation, microwave irradiation and thermal treatment: a comparative study. Carbohydrate Polymers, 79(4), 1080–1085.
Yu, C.-J., Ko, C.-J., Hsieh, C.-H., Chien, C.-T., Huang, L.-H., Lee, C.-W., & Jiang, C.-C. (2014). Proteomic analysis of osteoarthritic chondrocyte reveals the hyaluronic acid-regulated proteins involved in chondroprotective effect under oxidative stress. Journal of Proteomics, 99, 40–53.
Ong, K. L., Anderson, A. F., Niazi, F., Fierlinger, A. L., Kurtz, S. M., & Altman, R. D. (2016). Hyaluronic acid injections in medicare knee osteoarthritis patients are associated with longer time to knee arthroplasty. The Journal of Arthroplasty, 31(8), 1667–1673.
Kretz, F. T. A., Limberger, I.-J., & Auffarth, G. U. (2014). Corneal endothelial cell coating during phacoemulsification using a new dispersive hyaluronic acid ophthalmic viscosurgical device. Journal of Cataract and Refractive Surgery, 40(11), 1879–1884.
Stead, R. E., Juma, Z., Turner, S., Jones, L. D., & Sung, V. C. T. (2016). A novel use of reticulated hyaluronic acid (Healaflow) for hypotony eyes in patients with uveitis. British Journal of Ophthalmology, 100(6), 727–730.
Su, Z., Ma, H., Wu, Z., Zeng, H., Li, Z., Wang, Y., Liu, G., Xu, B., Lin, Y., Zhang, P., & Wei, X. (2014). Enhancement of skin wound healing with decellularized scaffolds loaded with hyaluronic acid and epidermal growth factor. Materials Science and Engineering C, 44, 440–448.
Wu, Z., Tang, Y., Fang, H., Su, Z., Xu, B., Lin, Y., Zhang, P., & Wei, X. (2015). Decellularized scaffolds containing hyaluronic acid and EGF for promoting the recovery of skin wounds. Journal of Materials Science: Materials in Medicine, 26(1), 1–10.
Wiest, L., & Kerscher, M. (2008). Native hyaluronic acid in dermatology – results of an expert meeting. Journal der Deutschen Dermatologischen Gesellschaft, 6(3), 176–180.
Sundaram, H., Mackiewicz, N., Burton, E., Peno-Mazzarino, L., Lati, E., & Meunier, S. (2016). Pilot comparative study of the topical action of a novel, crosslinked resilient hyaluronic acid on skin hydration and barrier function in a dynamic, three-dimensional human explant model. Journal of Drugs in Dermatology, 15(4), 434–441.
Pires, A. M. B., Macedo, A. C., Eguchi, S. Y., & Santana, M. H. A. (2010). Microbial production of hyaluronic acid from agricultural resource derivatives. Bioresource Technology, 101(16), 6506–6509.
Yamada, T., & Kawasaki, T. (2005). Microbial synthesis of hyaluronan and chitin: new approaches. Journal of Bioscience and Bioengineering, 99(6), 521–528.
Liu, L., Liu, Y., Li, J., Du, G., & Chen, J. (2011). Microbial production of hyaluronic acid: current state, challenges, and perspectives. Microbial Cell Factories, 10, 99.
Vázquez, J. A., Montemayor, M. I., Fraguas, J., & Murado, M. A. (2010). Hyaluronic acid production by Streptococcus zooepidemicus in marine by-products media from mussel processing wastewaters and tuna peptone viscera. Microbial Cell Factories, 9, 46.
Pires, A. M. B., & Santana, M. H. A. (2010). Metabolic effects of the initial glucose concentration on microbial production of hyaluronic acid. Applied Biochemistry and Biotechnology, 162(6), 1751–1761.
Shah, M. V., Badle, S. S., & Ramachandran, K. B. (2013). Hyaluronic acid production and molecular weight improvement by redirection of carbon flux towards its biosynthesis pathway. Biochemical Engineering Journal, 80, 53–60.
Vázquez, J. A., Pastrana, L., Piñeiro, C., Teixeira, J. A., Pérez-Martín, R. I., & Amado, I. R. (2015). Production of hyaluronic acid by Streptococcus zooepidemicus on protein substrates obtained from Scyliorhinus canicula discards. Marine Drugs, 13(10), 6537–6549.
Pan, N. C., Vignoli, J. A., Baldo, C., Pereira, H. C. B., Silva, R. S. S. F., & Celligoi, M. A. P. C. (2015). Agroindustrial byproducts for the production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920. International Journal of Scientific & Technology Research, 4(04), 114–118.
Oliveira, A. H., Ogrodowski, C. C., Macedo, A. C., Santana, M. H. A., & Gonçalves, L. R. B. (2013). Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production. Brazilian Journal of Microbiology, 44(4), 1097–1104.
Amado, I. R., Vázquez, J. A., Pastrana, L., & Teixeira, J. A. (2016). Cheese whey: a cost-effective alternative for hyaluronic acid production by Streptococcus zooepidemicus. Food Chemistry, 198, 54–61.
UNICA (2016). Sugarcane, ethanol and sugar production - 2015/2016 harvest season. In Brazilian Sugarcane Industry Association - UNICA. Retrieved May 23, 2016, from http://www.unicadata.com.br
Li, H., Jiang, Z., Yang, X., Yu, L., Zhang, G., Wu, J., & Liu, X. (2015). Sustainable resource opportunity for cane molasses: use of cane molasses as a grinding aid in the production of Portland cement. Journal of Cleaner Production, 93, 56–64.
Xu, S., Hao, N., Xu, L., Liu, Z., Yan, M., Li, Y., & Ouyang, P. (2015). Series fermentation production of ornithine and succinic acid from cane molasses by Corynebacterium glutamicum. Biochemical Engineering Journal, 99, 177–182.
Tyagi, N., & Suresh, S. (2016). Production of cellulose from sugarcane molasses using Gluconacetobacter intermedius SNT-1: optimization & characterization. Journal of Cleaner Production, 112, 71–80.
Ai, H., Liu, M., Yu, P., Zhang, S., Suo, Y., Luo, P., Li, S., & Wang, J. (2015). Improved welan gum production by Alcaligenes sp. ATCC31555 from pretreated cane molasses. Carbohydrate Polymers, 129, 35–43.
Ruiz, S. P., Martinez, C. O., Noce, A. S., Sampaio, A. R., Baesso, M. L., & Matioli, G. (2015). Biosynthesis of succinoglycan by Agrobacterium radiobacter NBRC 12665 immobilized on loofa sponge and cultivated in sugar cane molasses. Structural and rheological characterization of biopolymer. Journal of Molecular Catalysis B: Enzymatic, 122, 15–28.
Oliveira, M. R., Silva, R. S. S. F., Buzato, J. B., & Celligoi, M. A. P. C. (2007). Study of Levan production by Zymomonas mobilis using regional low-cost carbohydrate sources. Biochemical Engineering Journal, 37(2), 177–183.
Treichel, H., Mazutti, M. A., Maugeri Filho, F., & Rodrigues, M. I. (2009). Technical viability of the production, partial purification and characterisation of inulinase using pretreated agroindustrial residues. Bioprocess and Biosystems Engineering, 32(4), 425–433.
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28(3), 350–356.
Pan, N. C., Vignoli, J. A., Baldo, C., Pereira, H. C. B., Silva R. S. dos, S. F., & Celligoi, M. A. P. C (2015). Effect of fermentation conditions on the production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920. Acta Scientiarum. Biological Sciences, 37(4), 411–417.
Pires, A. M. B., Eguchi, S. Y., & Santana, M. H. A. (2010). The influence of mineral ions on the microbial production and molecular weight of hyaluronic acid. Applied Biochemistry and Biotechnology, 162(8), 2125–2135.
Tlapak-Simmons, V. L., Baron, C. A., & Weigel, P. H. (2004). Characterization of the purified hyaluronan synthase from Streptococcus equisimilis. Biochemistry, 43(28), 9234–9242.
Gomaa, E. Z. (2014). Production of polyhydroxyalkanoates (PHAs) by Bacillus subtilis and Escherichia coli grown on cane molasses fortified with ethanol. Brazilian Archives of Biology and Technology, 57(February), 145–154.
Chong, B. F., & Nielsen, L. K. (2003). Aerobic cultivation of Streptococcus zooepidemicus and the role of NADH oxidase. Biochemical Engineering Journal, 16(2), 153–162.
Armstrong, D. C., & Johns, M. R. (1997). Culture conditions affect the molecular weight properties of hyaluronic acid produced by Streptococcus zooepidemicus. Applied and Environmental Microbiology, 63(7), 2759–2764.
Lai, Z.-W., Rahim, R. A., Ariff, A., & Mohamad, R. (2011). Medium formulation and impeller design on the biosynthesis of high molecular weight hyaluronic acid by Streptococcus zooepidemicus ATCC 39920. African Journal of Microbiology Research, 5(15), 2114–2123.
Im, J.-H., Song, J.-M., Kang, J.-H., & Kang, D.-J. (2009). Optimization of medium components for high-molecular-weight hyaluronic acid production by Streptococcus sp. ID9102 via a statistical approach. Journal of Industrial Microbiology & Biotechnology, 36(11), 1337–1344.
Amrane, A., & Prigent, Y. (1994). Lactic acid production from lactose in batch culture : analysis of the data with the help of a mathematical model; relevance for nitrogen source and preculture assessment. Applied Microbiology and Biotechnology, 40, 644–649.
Haaland, P. D. (1989). Experimental design in biotechnology. (Marcell Dekker, Ed.). NY: CRC Press.
Armstrong, D. C., Cooney, M. J., & Johns, M. R. (1997). Growth and amino acid requirements of hyaluronic-acid-producing Streptococcus zooepidemicus. Applied Microbiology and Biotechnology, 47(3), 309–312.
Gao, H.-J., Du, G.-C., & Chen, J. (2006). Analysis of metabolic fluxes for hyaluronic acid (HA) production by Streptococcus zooepidemicus. World Journal of Microbiology and Biotechnology, 22(4), 399–408.
Gamboa-Suasnavart, R. A., Marín-Palacio, L. D., Martínez-Sotelo, J. A., Espitia, C., Servín-González, L., Valdez-Cruz, N. A., & Trujillo-Roldán, M. A. (2013). Scale-up from shake flasks to bioreactor, based on power input and Streptomyces lividans morphology, for the production of recombinant APA (45/47 kDa protein) from Mycobacterium tuberculosis. World Journal of Microbiology and Biotechnology, 29(8), 1421–1429.
Izawa, N., Hanamizu, T., Sone, T., & Chiba, K. (2010). Effects of fermentation conditions and soybean peptide supplementation on hyaluronic acid production by Streptococcus thermophilus strain YIT 2084 in milk. Journal of Bioscience and Bioengineering, 109(4), 356–360.
Liu, L., Wang, M., Du, G., & Chen, J. (2008). Enhanced hyaluronic acid production of Streptococcus zooepidemicus by an intermittent alkaline-stress strategy. Letters in Applied Microbiology, 46(3), 383–388.
Sun, X., Wang, Z., Bi, Y., Wang, Y., & Liu, H. (2015). Genetic and functional characterization of the hyaluronate lyase HylB and the Beta-N-Acetylglucosaminidase HylZ in Streptococcus zooepidemicus. Current Microbiology, 70(1), 35–42.
Rangaswamy, V., & Jain, D. (2008). An efficient process for production and purification of hyaluronic acid from Streptococcus equi subsp. zooepidemicus. Biotechnology Letters, 30(3), 493–496.
Lai, Z.-W., Rahim, R. A., Ariff, A. B., & Mohamad, R. (2012). Biosynthesis of high molecular weight hyaluronic acid by Streptococcus zooepidemicus using oxygen vector and optimum impeller tip speed. Journal of Bioscience and Bioengineering, 114(3), 286–291.
Jeong, E., Shim, W. Y., & Kim, J. H. (2014). Metabolic engineering of Pichia pastoris for production of hyaluronic acid with high molecular weight. Journal of Biotechnology, 185, 28–36.
Guillaumie, F., Furrer, P., Felt-Baeyens, O., Fuhlendorff, B. L., Nymand, S., Westh, P., Gurny, R., & Schwach-Abdellaoui, K. (2010). Comparative studies of various hyaluronic acids produced by microbial fermentation for potential topical ophthalmic applications. Journal of Biomedical Materials Research - Part A, 92(4), 1421–1430.
Gilli, R., Kacuráková, M., Mathlouthi, M., Navarini, L., & Paoletti, S. (1994). FTIR studies of sodium hyaluronate and its oligomers in the amorphous solid phase and in aqueous solution. Carbohydrate Research, 263(2), 315–326.
Wu, Y. (2012). Preparation of low-molecular-weight hyaluronic acid by ozone treatment. Carbohydrate Polymers, 89(2), 709–712.
El-Safory, N. S., & Lee, C.-K. (2010). Cytotoxic and antioxidant effects of unsaturated hyaluronic acid oligomers. Carbohydrate Polymers, 82(4), 1116–1123.
Kim, J. K., Srinivasan, P., Kim, J. H., Choi, J., Park, H. J., Byun, M. W., & Lee, J. W. (2008). Structural and antioxidant properties of gamma irradiated hyaluronic acid. Food Chemistry, 109(4), 763–770.
Ke, C., Sun, L., Qiao, D., Wang, D., & Zeng, X. (2011). Antioxidant acitivity of low molecular weight hyaluronic acid. Food and Chemical Toxicology, 49(10), 2670–2675.
Campo, G. M., Avenoso, A., Campo, S., D’Ascola, A., Ferlazzo, A. M., & Calatroni, A. (2004). The antioxidant and antifibrogenic effects of the glycosaminoglycans hyaluronic acid and chondroitin-4-sulphate in a subchronic rat model of carbon tetrachloride-induced liver fibrogenesis. Chemico-Biological Interactions, 148(3), 125–138.
Acknowledgements
The authors thank Coordination for the Improvement of Higher Education Personnel (CAPES – Brazil) for financial support, Dr. Dionisio Borsato from Londrina State University for support with the statistical analysis, and the Laboratory of Spectroscopy (SPEC) – State University of Londrina for the analyses.
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Pan, N.C., Pereira, H.C.B., da Silva, M.d.L.C. et al. Improvement Production of Hyaluronic Acid by Streptococcus zooepidemicus in Sugarcane Molasses. Appl Biochem Biotechnol 182, 276–293 (2017). https://doi.org/10.1007/s12010-016-2326-y
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DOI: https://doi.org/10.1007/s12010-016-2326-y