Agroindustrial Byproducts For The Production Of Hyaluronic Acid By Streptococcus Zooepidemicus ATCC 39920
Nicole Caldas Pan, Josiane Alessandra Vignoli, Cristiani Baldo, Hanny Cristina Braga Pereira, Rui Sérgio dos Santos Ferreira da Silva, Maria Antonia Pedrine Colabone Celligoi
Index Terms: hyaluronic acid, organic acid, soy protein, Streptococcus zooepidemicus, sugarcane molasses.
Abstract: Agroindustrial derivatives are alternative nutritional sources employed in bioprocesses that reduce costs and corroborate with social sustainability. In this study, alternative carbon (sugarcane juice, sugarcane molasses and soy molasses) and nitrogen sources (corn steep liquor, soy protein and whey protein) were evaluated for hyaluronic acid production by Streptococcus zooepidemicus ATCC 39920. The medium containing sugarcane molasses archived high yield of hyaluronic acid (0.066 g.g-1), when compared to the medium composed of glucose or sucrose. The replacement of yeast extract by soy protein was also effective for the production of the polymer resulting in 0.219 g.L-1. In general, the organic acids production was also evaluated and the results showed that the main metabolic products were lactate. In contrast, the acetate synthesis was detected only in the medium containing yeast extract. This study showed that sugarcane molasses is a promising carbon source for the hyaluronic acid production. This is the first study in which a culture media containing sugarcane molasses, a cheap substrate extensively produced in Brazil, has been successfully used for the microbial hyaluronic acid production.
 P.M. Meyer, P.H.M. Rodrigues and D.D. Millen, “Impact of Biofuel Production in Brazil on the Economy, Agriculture, and the Environment,” Anim. Front., vol. 3, pp. 28–37, 2013.
 C.R. Soccol and L.P.S. Vandenberghe, “Overview of Applied Solid-State Fermentation in Brazil,” Biochem. Eng. J., vol. 13, pp. 205–218, 2003.
 D. Zhang, X. Feng, Z. Zhou, Y. Zhang and H. Xu, “Economical Production of Poly(γ-glutamic acid) Using Untreated Cane Molasses and Monosodium Glutamate Waste Liquor by Bacillus subtilis NX-2,” Bioresour. Technol., vol. 114, pp. 583–588, 2012.
 E.J Vandamme, “Agro-Industrial Residue Utilization for Industrial Biotechnology Products,” Biotechnology for Agro-Industrial Residues Utilization, P. S. Nigam and A. Pandey, eds., Netherlands, Dordrecht: Springer, pp. 3 – 11, 2009.
 L.J. Benedini and M.H.A. Santana, “Effects of Soy Peptone on the Inoculum Preparation of Streptococcus zooepidemicus for Production of Hyaluronic Acid,” Bioresour. Technol., vol. 130, pp. 798–800, 2013.
 G.-Y. Lee, “Effect of Non-Animal-Derived Nitrogen Sources on the Production of Hyaluronic Acid by Streptococcus sp. KL0188,” J. Korean Soc. Appl. Biol. Chem., vol. 52, pp. 283–289, 2009.
 B.F. Chong, L.M. Blank, R. Mclaughlin and L.K Nielsen., “Microbial Hyaluronic Acid Production,” Appl. Microbiol. Biotechnol., vol. 66, pp. 341–351, 2005.
 G. Kogan, L. Soltés, R. Stern and P. Gemeiner, “Hyaluronic Acid: a Natural Biopolymer with a Broad Range of Biomedical and Industrial Applications,” Biotechnol. Lett., vol. 29, pp. 17–25, 2007.
 A.M.B. Pires, A.C. Macedo, S.Y. Eguchi and M.H.A. Santana, “Microbial Production of Hyaluronic Acid from Agricultural Resource Derivatives,” Bioresour. Technol., vol. 101, pp. 6506–6509, 2010.
 E. Marcellin, W. Chen and L.K. Nielsen “Microbial Hyaluronic Acid Biosynthesis,” Microbial Production of Biopolymers and Polymer Precursors: Applications and Perspectives, B.H.A. Rehm, ed., Norfolk: Caister Academic Press, pp. 163 –180, 2009.
 D.C. Armstrong, M.J. Cooney and M.R. Johns, “Growth and Amino Acid Requirements of Hyaluronic-Acid-Producing Streptococcus zooepidemicus,” Appl. Microbiol. Biotechnol., vol. 47, pp. 309–312, 1997.
 L. Liu, Y. Liu, J. Li, G. Du and J. Chen, “Microbial Production of Hyaluronic Acid: Current State, Challenges, and Perspectives,” Microb. Cell Fact., vol. 10:99, 2011.
 J.A. Vázquez, M.I. Montemayor, J. Fraguas and M.A. Murado, “Hyaluronic Acid Production by Streptococcus zooepidemicus in Marine By-Products Media from Mussel Processing Wastewaters and Tuna Peptone Viscera” Microb. Cell Fact., vol. 9:46, 2010.
 AOAC (Association of Official Agricultural Chemists), “Official Methods of Analysis,” AOAC, Washington DC, 1995.
 O.H. Lowry, N.J. Rosebrough, A.L. Farr and R.J. Randall, “Protein Measurement with the Folin Phenol Reagent,” J. Biol. Chem., vol. 193, pp. 265–275, 1951.
 T.M.C.C. Filisetti-Cozzi and N.C. Carpita, “Measurement of Uranic Acids without Interference from Neutral Sugars,” Anal. Biochem., vol. 197, pp. 157–162, 1991.
 M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers and F. Smith, “Colorimetric Method for Determination of Sugars and Related Substances,” Anal. Chem., vol. 28, pp. 350–356, 1956.
 M.R. Oliveira, R.S.S.F. Silva, J.B. Buzato and M.A.P.C. Celligoi, “Study of Levan Production by Zymomonas mobilis Using Regional Low-Cost Carbohydrate Sources,” Biochem. Eng. J., vol. 37, pp. 177–183, 2007.
 K. Xu and P. Xu, “Efficient Production of L-lactic Acid Using Co-feeding Strategy Based on Cane Molasses/Glucose Carbon Sources,” Bioresour. Technol., vol. 153, pp. 23–29, 2014.
 A.C. Macedo and M.H.A. Santana, “Hyaluronic Acid Depolymerization by Ascorbate-Redox Effects on Solid State Cultivation of Streptococcus zooepidemicus in Cashew Apple Fruit Bagasse,” World J. Microbiol. Biotechnol., vol. 28, pp. 2213–2219, 2012.
 F.B. Chong and L.K. Nielsen, “Aerobic Cultivation of Streptococcus zooepidemicus and the Role of NADH Oxidase,” Biochem. Eng. J., vol. 16, pp. 153–162, 2003.
 K. Abbe, S. Takahashi and T. Yamada, “Involvement of Oxygen-Sensitive Pyruvate Formate-Lyase in Mixed-Acid Fermentation by Streptococcus mutans Under Strictly Anaerobic Conditions,” J. Bacteriol., vol. 152, pp. 175–182, 1982.
 M. V. Shah, S.S. Badle and K.B. Ramachandran, “Hyaluronic Acid Production and Molecular Weight Improvement by Redirection of Carbon Flux Towards its Biosynthesis Pathway,” Biochem. Eng. J., vol. 80, pp. 53–60, 2013.
 A. Amrane and Y. Prigent “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,” Appl. Microbiol. Biotechnol., vol. 40, pp. 644–649, 1994.
 Z.-W. Lai, R.A. Rahim, A. Ariff and R. Mohamad, “Medium Formulation and Impeller Design on the Biosynthesis of High Molecular Weight Hyaluronic Acid by Streptococcus zooepidemicus ATCC 39920,” African J. Microbiol. Res., vol. 5, pp. 2114–2123, 2011.