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Statistical Optimization of Process Parameters for Bioplastic (PHA) Production by Bacillus subtilis NCDC0671 Using Orange Peel-Based Medium

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Abstract

Systematic and sequential optimization strategy based on statistical methodology involving one factor at a time approach was used to enhance PHA production by Bacillus subtilis NCDC0671 using orange peel hydrolysate-based medium in shake flask cultures. Medium components exerting a significant influence in PHA production were screened out using the Plackett–Burman design. Following Box–Behnken method, three significant factors influencing PHA production were identified as beef extract, sodium chloride and incubation period. The optimum concentration and interaction effect of these three components were determined by RSM based on Box–Behnken design. The model developed was validated, and the optimum medium containing 2.23 (g/L) beef extract, 0.72 (g/L) sodium chloride and 48 h of incubation period led to maximum PHA production of 5.09 g/L, which was 2.49-fold higher than the unoptimized medium. The ANOVA results established the significance of the model (p < 0.05). The present investigation innovatively developed a low-cost fermentation medium utilizing orange peels to produce considerable level of PHA.

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References

  • AndreeBen B, Lange AB, Robenek H, Steinbuchel A (2010) Conversion of glycerol to poly 3 hdroxy propionate in recombinant E. coli. Appl Environ Microbiol 76:622–626

    Article  Google Scholar 

  • Anjum MF, Tasadduq I, Al-Sultan K (1997) Response surface methodology: a neural network approach. Eur J Oper Res 101:65–73

    Article  Google Scholar 

  • AOAC (2005) Official methods of analysis of A.O.A.C., 18th edn. In: Helirich K (ed) Association of Official Analytical Chemists, Inc., Arlington, VA

  • APHA, AWWA, WEF (2005) Standard methods for the examination of water and waste water, 21st edn. Washington, D.C

  • Benoit TG, Wilson GR, Baugh CL (1990) Fermentation during growth and sporulation of Bacillus thuringiensis HD-1. Lett Appl Microbiol 10:15–18

    Article  Google Scholar 

  • Borah B, Thakur PS, Nigam JN (2002) The influence of nutritional and environmental conditions on the accumulation of poly-hydroxybutyrate in Bacillus mycoides RLJ B-017. J Appl Microbiol 92:776–783

    Article  Google Scholar 

  • Box GEP, Behnken DW (1960) Some new three level designs for the study of quantitative variables. Technometrics 2:455–475

    Article  MathSciNet  Google Scholar 

  • Egbuonu ACC, Omodamiro OD, Odo CE, Uroko RI (2016) Some antinutritive and antioxidative properties of pulverized Citrus sinensis (sweet orange) peels and seeds. J Sci Res Rep 10:1–9

    Google Scholar 

  • Elsayed NS, Aboulwafa M, Aboshanab K, Hassouna N (2013) PHB production in Azomonas, Acinteobacter and Bacillus species: isolation, screening and identification. Arch Clin Microbiol 4:5025–5035

  • Hedge JE, Hofreiter BT (1962) Carbohydrate chemistry. In: Whistler RL, Be Miller JN (eds) vol 17, Academic Press, New York

  • Hegazy AE, Ibrahium MI (2012) Antioxidant activities of orange peel extracts. World Appl Sci J 18:684–688

    Google Scholar 

  • Karoui IJ, Marzouk B (2013) Characterization of bioactive compounds in Tunisian bitter orange (Citrus aurantium L.) peel and juice and determination of their antioxidant activities. Bio Med Res Int 1:1–12

  • Khuri AI, Cornell JA (1987) Response surfaces: designs and analyses. Marcel Dekker Inc., New York

    MATH  Google Scholar 

  • Law JH, Slepecky RA (1961) Assay of poly-β-hydroxybutyric acid. J Bacteriol 82:33–36

    Google Scholar 

  • Li XY, Liu ZQ, Chi ZM (2008) Production of phytase by a marine yeast Kodamaea ohmeri BG3 in an oats medium: optimization by response surface methodology. Bioresour Technol 99:6386–6390

    Article  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Mandalari G, Bennett RN, Bisignano G, Saija A, Dugo G, Faulds CB, Waldron KW (2006) Characterization of flavonoids and pectin from bergamot (Citrus bergamia Risso) peel, a major byproduct of essential oil extraction. J Agric Food Chem 54:197–203

    Article  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–427

    Article  Google Scholar 

  • Muralidharan R, Radha KV (2014) A kinetic study on polyhydroxybutyrate production on nitrogen limited medium using Bacillus subtilis MTCC9763 through a two stage cultivation strategy. J Environ Biol 36:537–542

    Google Scholar 

  • Myers RH, Montgomery DC (1995) Response surface methodology: process and product optimization using designed experiments. Wiley, New York

    MATH  Google Scholar 

  • Nam DH, Ryu DDY (1985) Relationship between butirosin biosynthesis and sporulation in Bacillus circulans. Antimicrob Agents Chemother 27:789–801

    Article  Google Scholar 

  • Oikeh EI, Oriakhi K, Omoregie ES (2013) Proximate analysis and phytochemical screening of citrus sinesis fruit waste. Bioscientist 1:164–170

    Google Scholar 

  • Omar S, Rayes A, Eqaab A, Steinbuchel A (2001) Optimization of cell growth and poly (3-hydroxybutyrate) accumulation on date syrup by a Bacillus megaterium strain. Biotechnol Lett 23:1119–1123

    Article  Google Scholar 

  • Plackett RL, Burman JP (1946) The design of optimum multifactorial experiments. Biometrica 33:305–325

    Article  MathSciNet  Google Scholar 

  • Prasanna T, Ajay Babu P, Dhanavara L, Chakrapani R, Ramachandra Rao CSV (2011) Production of poly (3-hydroxybutyrate) by Bacillus species isolated from soil. J Pharma Res Rev 1:15–18

    Google Scholar 

  • Pumiput P, Chuntranuluck S, Kitpreechavanich V, Punsuron V, Vaithanomstat P (2008) Production process of hydrolysate from steam explosion of oil trunk for xylitol fermentation. Kasetsart J (National Science) 42:73–78

    Google Scholar 

  • Rivas B, Torrado A, Torre P, Converti A, Domínguez JM (2008) Submerged citric acid fermentation on orange peel autohydrolysate. J Agric Food Chem 56:2380–2387

    Article  Google Scholar 

  • Ruchi G, Anshu G, Khare SK (2008) Lipase from solvent tolerant Pseudomonas aeruginosa strain: production optimization by response surface methodology and application. Bioresour Technol 99:4796–4802

    Article  Google Scholar 

  • Santhanam A, Sasidharan S (2010) Microbial production of PHA from Alcaligens spp. and Pseudomonas oleovorans using different carbon sources. Afr J Biotechnol 9:3144–3150

    Google Scholar 

  • Sathiyanarayanan G, Saibaba G, Kiran GS, Selvin J (2013) A statistical approach for optimization of polyhydroxybutyrate production by marine Bacillus subtilis MSBN17. Int J Biol Macromol 59:170–177

    Article  Google Scholar 

  • Shang L, Jiang M, Yun ZH, Yan HQ, Chang HN (2008) Mass production of medium-chain-length poly (3-hydroxyalkanoates) from hydrolyzed corn oil by fed-batch culture of Pseudomonas putida. World J Microbiol Biotechnol 24:2783–2787

    Article  Google Scholar 

  • Sing G, Kumari A, Mittal A, Goel V, Yadav A, Aggarwal NK (2013) Cost effective production of poly-β-hydroxybutyrate by Bacillus subtilis NG05 using sugar industry waste water. Bio Med Res Int l:1–10

  • Torrado AM, Cortes S, Salgado JM, Max B, Rodriguez N, Bibbins BP, Dominguez JM (2011) Citric acid production from orange peel wastes by solid-state fermentation. Braz J Microbiol 42:394–409

    Article  Google Scholar 

  • Tsuge T (2002) metabolic improvements and use of inexpensive carbon sources in microbial production of polyhydroxyalkanoates. J Biosci Bioeng 6:579–584

    Article  Google Scholar 

  • Valappil SP, Bocaccini AR, Bucke C, Roy I (2007) Polyhydroxyalkanoates in gram positive bacteria: insight fron the genera Bacillus and sstreptomyces. Antonie Van Leeuwenhoek 91:1–17

    Article  Google Scholar 

  • Vijyaendra SVN, Rastogi NK, Shyamala TR, Anil kumar PK, Kshama L, Joshi GJ (2007) Optimization of polyhydroxybutyrate production by Bacillus sp CFR256 with corn steep liquor as nitrogen source. Indian J Microbiol 47:170–175

    Article  Google Scholar 

  • Westers H, Dorenbos R, Van Dijil JM, Kabel J, Flanagan T, Devine KM, Jude F, Seror SJ, Beekman AC, Darmon E, Eschevins C, de Jong A, Bron S, Kuipers OP, Albertini AM, Antelmann H, Hecker M, Zamboni N, Sauer U, Bruand C, Ehrlich DS, Alonso JC, Salas M, Quax WJ (2003) Genome engineering reveals large dispensable regions in Bacillus subtilis. Mol Biol Evol 20:2076–2090

    Article  Google Scholar 

  • Yamane T, Fukunage M, Lee YW (1996) Increased PHB productivity by high cell density fed batch culture of Alcaligens latus a growth associated PHB producer. Biotechnol Bioeng 50:197–202

    Article  Google Scholar 

  • Yu JC, Liu Q, Liu XD, Sun Q, Yan JF (2008) Effect of liquid culture requirements on antifungal antibiotic production by Streptomyces rimousus MY02. Bioresour Technol 99:2087–2091

    Article  Google Scholar 

Download references

Acknowledgements

This research did not receive any specific Grant from funding agencies in the public, commercial, or not-for-profit sectors. We would like to acknowledge the Department of Microbial Biotechnology for providing necessary facilities for carrying out this research work.

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MU, VMM and TB have equally contributed to the planning, execution of work and preparation of manuscript. KP has served as supervisor and guide for the research work and manuscript preparation.

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Correspondence to Mridul Umesh.

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Umesh, M., Mani, V.M., Thazeem, B. et al. Statistical Optimization of Process Parameters for Bioplastic (PHA) Production by Bacillus subtilis NCDC0671 Using Orange Peel-Based Medium. Iran J Sci Technol Trans Sci 42, 1947–1955 (2018). https://doi.org/10.1007/s40995-017-0457-9

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