• Natarajan K. T John College of Pharmacy, Department of Pharmaceutical Biotechnology, Bengaluru.
  • Vineeth Chandy T John College of Pharmacy, Department of Pharmaceutical Biotechnology, Bengaluru.
  • Sirajudeen M. A. Ultra College of Pharmacy, Department of Pharmaceutical Biotechnology, Madurai


Objective: This study was done to optimize the production parameters involved in the isolation of hyaluronan (HA) from UV mutated a clinical strain of Klebsiella pneumonia (M 3020).

Methods: Glucose and nitrogen enriched media (D-glucose, L-glutamic acid, and peptone) were utilized to cultivate the clinical isolate K. pneumoniae. The strain was Ultra Violet (UV) radiation mutated (254 nm, 25 min) and HA production was optimized by parameters such as pH and temperature. The isolated HA from the fermented broth was subjected to purification by isopropyl alcohol and silica gel and further dried by lyophilization. Produced HA was confirmed with UV and Fourier Transform Infra-Red (FT-IR) spectroscopy.

Results: UV treated strain at 254 nm for 25 min predominantly produce a high quantity of HA (3.5 g/l) in 37 °C, 300 rpm and pH 6.8 at 24 h run. UV and IR spectrum of produced HA showed strong similarity with the standard hyaluronan.

Conclusion: To conclude, high quantity and quality of HA can be isolated from mutated clinical strains of K. pneumoniae.

Keywords: HA, Klebsiella pneumoniae, Mutation, Enriched media


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1. Scott JE, Christine C, Andy B, Yuan Chen. Secondary and tertiary structures of HA in aqueous solution, investigated by rotary shadowing electron microscopy and computer simulation. Biochem J 1991;274:699-705.
2. Abigail C, Fabien G, Jerome FLD. The major surface associated saccharides of Klebseilla pneumoniae contribute to host cell association. PloS One 2008;3:381-7.
3. Fedelic Ashish Toppo, Rajesh Singh Pawar. Novel drug delivery strategies and approaches for wound healing management. J Crit Rev 2015;2:12-20.
4. Santhi Priya Nagam A, Naga Jyothi J, Poojitha, Santhosh Aruna, Rama Rao Nadendla A. Comprehensive review on hydrogels. Int J Curr Pharm Res 2016;8:19-23.
5. Necas J, Bartosikova L, Brauner P, J Kolar. Hyaluronic acid: a review. Veterina Med 2008;53:397-11.
6. Pan NC, Vignoli JA, Baldo C. Effect of fermentation conditions on the production of HA by Streptococcus zooepidemicus ATCC 39920. Acta Sci 2015;37:411-7.
7. Chong BF, Blank M, Mclaughlin R, Nielsen K. Microbial HA production. Appl Microbiol Biotechnol 2005;66:341-35.
8. David C, Johns R. Culture conditions affect the molecular weight properties of HA produced by Streptococcus zooepidemicus. Appl Environ Microbiol 1997;63:2759–64.
9. Bill W, Regine B, Steve V, Maria T, Tia H, Alan S, et al. HA production in Bacillus subtilis. Appl Environ Microbiol 2005;3:747–52.
10. Mandeep Kaur, Guhan Jayaraman. HA production and the molecular weight is enhanced in pathway-engineered strains of lactate dehydrogenase-deficient Lactococcus lactis. Metabol Eng Comm 2016;3:15–23.
11. Takahashi M, Yoshida K, San Clemente CL. Relation of colonial morphologies in soft agar to morphological and biological properties of the K-9 strain of Klebsiella pmeumoniae and its variants. Canadian J Microbiol 1977;23:448–51.
12. Bitter T, Muir H. A modified uronic-acid-carbazole reaction. Anal Biochem 1962;4:320-34.
13. Eric Fournier. Colorimetric quantification of carbohydrates. Curr Protoc Food Anal Chem 2001;E1:1-8.
14. Thonard JC, Migliore XA, R Blustein. Isolation of HA from broth cultures of Streptococci. J Biol Chem 1964;239:726-8.
15. Lee HG, Cowman MK. An agarose gel electrophoretic method for analysis of HA molecular weight distribution. Anal Biochem 1994;219:278-87.
16. Johns MR, Goh LT, Oeggerli A. Effect of pH, agitation and aeration on HA production by Streptococcus zooepidemicus. Biotechnol Lett 1994;16:507-12.
17. Shardul Bhilocha, Ripal Amin, Monika Pandya. Agarose and polyacrylamide gel electrophoresis methods for molecular mass analysis of 5–500 kDa HA. Anal Biochem 2011;417:41–9.
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How to Cite
K., N., V. Chandy, and S. M. A. “HYALURONAN-A NOVEL POLYMER ISOLATED FROM MUTATED CLINICAL BACTERIAL ISOLATE”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 1, Jan. 2018, pp. 208-10, doi:10.22159/ijpps.2018v10i1.22624.
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