Prerana Venkatachalam, Varalakshmi Kilingar Nadumane
Prerana Venkatachalam, Varalakshmi Kilingar Nadumane*
Department of Biotechnology, School of Sciences, JAIN (Deemed-to-be University), 18/3, 9th Main, 3rd block, Jayanagar, Bengaluru – 560011, India.
Volume - 14,
Issue - 1,
Year - 2021
As a pink coloured bioactive fraction, B5, produced by B. endophyticus JUPR15 was showing promising anti-cancer properties, response surface methodology (RSM) was employed to enhance its yield. Various carbon, nitrogen, amino acids, vitamins and metal ions along with varying pH, temperature and incubation time were screened independently for enhancing pigment and biomass yield. Central Composite Design (CCD) of RSM was applied to evaluate the optimal conditions for the chosen factors – glycerol, mannitol and malt extract, which were studied at five levels. The R2 value for biomass was 0.9874 and that for pigment yield was 0.9931. The observed responses were in close agreement with the predicted responses. Through CCD, a 14.31-fold (12.02 g/L) increase in biomass and 7.77-fold (202.2 mg/L) increase in pigment yields were achieved as opposed to the 0.84 g/L and 26.03 mg/L of biomass and pigment yields obtained through control conditions. The model was found to be significant (P<0.01) for both the responses. Besides this, the addition of mannitol alone gave 191.36 mg/L of pigment, closer to that obtained via optimized conditions. It can be concluded that both single factor and RSM can be employed to increase the anti-cancer pigment production.
Cite this article:
Prerana Venkatachalam, Varalakshmi Kilingar Nadumane. Enhanced Production of an Anti-Cancer Pigment from Bacillus endophyticus JUPR15: Single Factor System Vs RSM. Research J. Pharm. and Tech. 2021; 14(1):153-161. doi: 10.5958/0974-360X.2021.00027.5
Prerana Venkatachalam, Varalakshmi Kilingar Nadumane. Enhanced Production of an Anti-Cancer Pigment from Bacillus endophyticus JUPR15: Single Factor System Vs RSM. Research J. Pharm. and Tech. 2021; 14(1):153-161. doi: 10.5958/0974-360X.2021.00027.5 Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2021-14-1-27
1. Lyngwi NA and Joshi SR. Economically important Bacillus and related genera: a mini review. In: Sen A, editor. Biology of useful plants and microbes. New Delhi: Narosa Publishing House. 2014; pp. 33-43.
2. Lim SH, Choi JS and Park EY. Microbial production of riboflavin using riboflavin overproducers, Ashbya gossypii, Bacillus subtilis, and Candida famata: an overview. Biotechnol Bioprocess Eng. 2001; 6(2): 75-88.
3. Perez-Fons L, Steiger S, Khaneja R and Bramley PM. Identification and the developmental formation of carotenoid pigments in the yellow/orange Bacillus spore-formers. Biochim Biophys Acta. 2011; 1811(3): 177–185.
4. Ouhib O, Clavel T and Schmitt P. The production of Bacillus cereus enterotoxins is inﬂuenced by carbohydrate and growth rate. Curr Microbiol. 2006; 53(3): 222–226.
5. Iwasaki T, Ishibashi J, Tanaka H and Sato M. Selective cancer cell cytotoxicity of enantiomeric 9- mer peptides derived from beetle defensins depends on negatively charged phosphatidylserine on the cell surface. Peptides. 2009; 30(4): 660–668.
6. Berdy J. Bioactive microbial metabolites. J Antibiot (Tokyo). 2005; 58(1): 1–26.
7. Sen T, Barrow CJ and Deshmukh SK. Microbial pigments in the food industry-Challenges and the way forward. Front Nutr. 2019; 6: article 7.
8. Rao MPN, Xiao M and Li W-J. Fungal and bacterial pigments: Secondary metabolites with wide applications. Front Microbiol. 2017; 8:1113.
9. Manikprabhu D and Lingappa K. γ Actinorhodin a natural and attorney source for synthetic dye to detect acid production of fungi. Saudi J Biol Sci. 2013; 20(2):163–168.
10. Tuli HS, Chaudhary P, Beniwal V and Sharma AK. Microbial pigments as natural color sources: current trends and future perspectives. J Food Sci Technol. 2015; 52(8): 4669–4678.
11. Venil CK, Zakaria ZA and Ahmad WA. Bacterial pigments and their applications. Process Biochem. 2013; 48(7):1065–1079.
12. Wang H, Ren Z, Li P and Gu Y. Improvement of the production of a red pigment in Penicillium sp. HSD07B synthesized during co-culture with Candida tropicalis. Bioresource Technol. 2011; 102(10): 6082-6087.
13. Wang H, Jiang P, Lu Y and Ruan Z. Optimization of culture conditions for violacein production by a new strain of Duganella sp. B2. Biochem Eng J. 2009; 44(2-3): 119–124.
14. Kennedy M and Krouse D. Strategies for improving fermentation medium performance: A review. J Ind Microbiol Biotechnol. 1999; 23(6): 456-475.
15. Nasri NMR and Razavi SH. Use of response surface methodology
in a fed-batch process for optimization of tricarboxylic acid cycle intermediates to achieve high levels of canthaxanthin from Dietzia natronolimnaea HS-1. J Biosci Bioeng. 2010; 109(4): 361-368.
16. Haaland PD. Experimental Design in Biotechnology. CRC Press, Taylor and Francis group. 1989.
17. Hamidi M, Abdin MZ, Nazemyieh H and Hejazi MA. Optimization of total carotenoid production by Halorubrum sp. TBZ126 using response surface methodology. J Microb Biochem Technol. 2014; 6(5): 286-294.
18. Polak-Berecka M, Wasko A, Kordowska-Wiater M and Podlesny M. Optimization of medium composition for enhancing growth of Lactobacillus rhamnosus PEN using response surface methodology. Pol J Microbiol. 2010; 59(2): 113-118.
19. Aneja KR. Cultivation techniques for isolation and enumeration of microorganisms. In: Experiments in Microbiology, Plant Pathology and Biotechnology, 4th edn. New Delhi: New Age International Publishers. 2003; pp. 154-188.
20. Venkatachalam P and Nadumane VK. Overexpression of p53 and Bax mediating apoptosis in cancer cell lines induced by a bioactive compound from Bacillus endophyticus JUPR15. Process Biochem. 2018; 73: 170-179.
21. Montgomery DC. Design and analysis of experiments, 8th edn. Wiley, Hoboken; 2013.
22. Sarvamangala D and Aparna SSV. Microbial Pigments – A short review. IOSR-JESTFT. 2016; 10(8): 1-7.
23. Dufosse L. Pigments. Microbial Encylopedia Microbiol. 2009; 4:457-471.
24. Heer K and Sharma S. Microbial pigments as a natural color: A review. Int J Pharm Sci Res. 2017; 8(5):1913-1922.
25. Khaneja R, Perez-Fons L, Fakhry S and Baccigalupi L. Carotenoids found in Bacillus. J Appl Microbiol. 2010; 108(6):1889–1902.
26. Zur Y, Gitelson AA, Chivkunova OB and Merzlyak MN. The spectral contribution of carotenoids to light absorption and reflectance in green leaves. Proceedings of the 2nd International Conference Geospatial Information in Agriculture and Forestry, Buena Vista, FL, v. 2. 2000.
27. Rodriguez-Amaya DB. A guide to carotenoid analysis in foods. Washington, DC: ILSI Press. 2001.
28. Kim HO, Lim JM, Joo JH and Kim SW. Optimization of submerged culture condition for the production of mycelial biomass and exopolysaccharides by Agrocybe cylindracea. Bioresour Technol. 2005; 96(10):1175-1182.
29. Chen XH, Lou WY, Zong MH and Smith TJ. Optimization of culture conditions to produce high yields of active Acetobacter sp. CCTCC M209061 cells for anti-Prelog reduction of prochiral ketones. BMC Biotechnol. 2011; 11:110.
30. Ram MS, Kumar NP, Bhaskar CVS and Audipudi AV. Optimization and characterization of intracellular orange fluorescent pigment from Bacillus endophyticus (AVP-9(Kf527823)). Int J Curr Pharm Res. 2017; 9: 67-74.
31. Ryan FJ. Selected methods of Neurospora genetics. Meth Med Res. 1950; 3: 51-75.
32. De Felice M, Levinthal M, Iaccarino M and Guardiola J. Growth inhibition as a consequence of antagonism between related amino acids: effect of valine in Escherichia coli. Microbiol Rev. 1979; 43(1): 42–58.
33. Kajikawa H, Mitsumori M and Ohmomo S. Stimulatory and inhibitory effects of protein amino acids on growth rate and efficiency of mixed ruminal bacteria. J Dairy Sci. 2002; 85(8): 2015–2022.
34. Lin ES and Sung SC. Cultivating conditions influence exopolysaccharide production by the edible basidiomycete Antrodia cinnamomea in submerged culture. Int J Food Microbiol. 2006; 108(2):182-187.
35. Cai CG and Zheng XD. Medium optimization for keratinase production in hair substrate by a new Bacillus subtilis KD-N2 using response surface methodology. J Ind Microbiol Biotechnol. 2009; 36(7): 875-883.