Poly-β-hydroxybutyrate production and management of cardboard industry effluent by new Bacillus sp. NA10

Abstract

Background

In the present study, we aim to utilize the ecological diversity of soil for the isolation and screening for poly β-hydroxybutyrate (PHB)-accumulating bacteria and production of cost-effective bioplastic using cardboard industry effluent.

Results

A total of 120 isolates were isolated from different soil samples and a total of 62 isolates showed positive results with Nile blue A staining, a specific dye for PHB granules and 27 isolates produced PHB using cardboard industry effluent. The selected isolate NA10 was identified as Bacillus sp. NA10 by studying its morphological, biochemical, and molecular characteristics. The growth pattern for the microorganism was studied by logistic model and exactly fitted in the model. A maximum cell dry weight (CDW) of 7.8 g l⁻¹ with a PHB concentration of 5.202 g l⁻¹ was obtained when batch cultivation was conducted at 37°C for 72 h, and the PHB content was up to 66.6% and productivity was 0.072 g l⁻¹ h⁻¹ in 2.0 L fermentor. Chemical characterization of the extracted PHB was done by H₁NMR, Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), Gas chromatography–mass spectrometry (GC-MS) analysis to determine the structure, melting point, and molecular mass of the purified PHB. The polymer sheet of extracted polymer was prepared by blending the polymer with starch for packaging applications.

Conclusions

The isolate NA10 can be a good candidate for industrial production of PHB from cardboard industry waste water cost-effectively and ecofriendly.

Keywords

Polyhydroxybutyrate (PHB) / Bioplastic / Cardboard industry waste water / Bacillus sp.

References

[1] Lazarevic D, Aoustin E, Buclet N, Brandt N. Plastic waste management in the context of a European recycling society: comparing results and uncertainties in a life cycle perspective. Resour Conserv Recycling vol, 2010, 55: 246-259.

[2] Flechter A. Plastics from bacteria and for bacteria: PHA as natural, biodegradable polyesters, 1993, New York: Springer.

[3] Nath A, Dixit M, Bandiya A. Enhanced PHB production and scale up studies using cheese whey in fed batch culture of Methylobacterium sp. ZP 24. Bioresource Technol, 2008, 99: 5749-5755.

[4] Anderson AJ, Dawes EA. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev, 1990, 54: 450-472.

[5] Yu J, Stahl H. Microbial utilization and biopolyester synthesis of bagasse hydrolysates. Bioresource Technol, 2008, 99: 8042-8048.

[6] Lee IY, Chang HN, Park YH. A simple method for recovery of microbial poly-3-hydroxybutyrate by alkaline solution treatment. J Microbiol Biotechnol, 1995, 5 4 238-240.

[7] Gao X, Chen JC, Wu Q, Chen GQ. Polyhydroxyalkanoates as a source of chemicals, polymers and biofuels. Curr Opin Biotechnol, 2011, 22: 768-774.

[8] Choi JI, Lee SY. Process analysis and economic evaluation for poly (3-hydroxybutyrate) production by fermentation. Bioprocess Eng, 1997, 17: 335-342.

[9] Yamane T, Chen XF, Ueda S. Growth associated production of poly(3-hydroxyvalerate) from n-pentanol by a methylotrophic bacterium, Paracoccus denitificans. Appl Environ Microbiol, 1996, 62: 380-384.

[10] Grothe E, Moo-Young M, Chisti Y. Fermentation optimization for the production of poly (ß-hydroxybutyric acid) microbial thermoplastic. Enzyme Microb Technol, 1999, 25: 132-141.

[11] Wen Q, Chen Z, Tian T, Chen W. Effects of phosphorus and nitrogen limitation on PHA production in activated sludge. J Environ Scien, 2010, 22 10 1602-1607.

[12] Ceyhan N, Ozdemir G. Polyhydroxybutyrate (PHB) production from domestic wastewater using Enterobacter aerogenes 12Bi strain. Afr J Microbiol Res, 2011, 5 6 690-702.

[13] Wong HH, Lee SY. Poly-(3-hydroxybutyrate) production from whey by high- density cultivation of recombinant Escherichia coli. Appl Microbiol Biotechnol, 1998, 50: 30-33.

[14] Rebah FB, Prévost D, Tyagi RD, Belbahri L. Poly-β-hydroxybutyrate production by fast-growing Rhizobia cultivated in sludge and in industrial wastewater. Appl Biochem Biotech, 2009, 158 1 155-163.

[15] Reddy VST, Thirumala M, Mahmood KS. Production of P HB and P(3HB-co-3HV) biopolymers of Bacillus megaterium strain OU303A isolated from municipal sewage sludge. W J Microbiol Biotechnol, 2009, 25: 391-397.

[16] Koller MB, C hiellini E, Fernandes EG, Horvat P, K utschera C, Hesse P, Braunegg G. Polyhydroxyalkanoate production from whey by Pseudomonas hydrogenovora. Bioresour Technol, 2008, 99 11 4854-4863.

[17] Bengtsson S, Werker A, Welander T. Production of polyhydroxyalkanoates by glycogen accumulating organisms treating a paper mill wastewater. Water Scien Technol, 2008, 58: 323-330.

[18] Du G, Yu J. Green technology for conversion of food scraps to biodegradable thermoplastic polyhydroxyalkanoates. Environ Sci Technol, 2002, 36: 5511-5516.

[19] Juan ML, Gonzalez LW, Walker GC. A novel screening method for isolating exopolysaccharide-deficient mutants. Appl Environ Microbiol, 1998, 64: 4600-4602.

[20] Spiekermann P, Rehm BH, Kalscheuer R, Baumeister D, Steinbüchel A. A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Arch Microbiol, 1999, 171 2 73-80.

[21] Singh G, Mittal A, Kumari A, Goel V, Aggarwal NK, Yadav A. Optimization of poly-β-hydroxybutyrate production from Bacillus species. Eur J Biol Scien, 2011, 3 4 112-116.

[22] Law J, Slepecky RA. Assay of poly-hydroxybutyric acid. J Bacteriol, 1961, 82: 52-55.

[23] Lee YU, Yoo YJ. Kinetics for the growth of Alcaligenes eutrophus and the biosynthesis of poly-β-hydoxybutyrate. Korean J Appl. Microbiol. Biotechnol, 1991, 19: 186-92.

[24] Painter PR, Marr AG. Mathematics of microbial populations. Ann Rev Microbiol, 1963, 22: 219-221.

[25] Levasseur M, Thompson PA, Harrison Paul J. Physiological acclimation of marine phytoplankton to different nitrogen sources. J Phycol, 1993, 29 5 587-595.

[26] Wang F, Lee SY. Poly(3-hydroxybutyrate) production with high productivity and high polymer content by a fed-batch culture of Alcaligenes latus under nitrogen limitation. Appl Environ Microbiol, 1997, 63 9 3703-3706.

[27] Du G, Yu J, Chen J, Lun S. Continuous production of poly-3-hydroxybutyrate by Ralstonia eutropha in a two stage culture system. J Biotechnol, 2001, 88: 59-65.

[28] Steinbüchel A. Perspectives for biotechnological production and utilization of biopolymers: metabolic engineering of polyhydroxyalkanoats biosynthesis pathways as a successful example. Macromol Bioscien, 2001, 1: 1-24.

[29] Teeka J, Cheng I, Xuehang T, Alissara R, Takaya H, Koichi Y, Masahiko S. Screening of PHA-producing bacteria using biodiesel-derived waste glycerol as a sole carbon source. J Water Environ Technol, 2010, 8: 371-381.

[30] Ramachandran H, Abdullah AA (2010) Isolation of PHA-producing bacteria from Malaysian Environment. Proceedings of the 7th IMT-GT UNINET and The 3rd International PSUUNS Conferences on Bioscience 178–179 Ramachandran H, Abdullah AA (2010) Isolation of PHA-producing bacteria from Malaysian Environment. Proceedings of the 7th IMT-GT UNINET and The 3rd International PSUUNS Conferences on Bioscience 178–179

[31] Kitamura S, Doi Y. Staining method of poly (3-hydroxyalkanoic acid) producing bacteria by Nile Blue. Biotechnol Tech, 1994, 8: 345-350.

[32] Salehizadeh H, Van Loosdrecht MCM. Production of polyhydroxyalkanoates by mixed culture: recent trends and biotechnological importance. Biotechnol Adv, 2004, 22: 261-279.

[33] Dias JM, Lemos PC, Serafim LS, Oliveira EC, Albuquerque M, Ramos M, Oliveira R, Reis MA. Recent advances in polyhydroxyalkanoate production by mixed aerobic cultures: from the substrate to the final product. Macromol Biosci, 2006, 6: 885-906.

[34] Ganzeveld KJ, Van Hagen A, Van Agteren MH, Koning DW, Uiterkamp AMJS. Upgrading of organic waste: production of the copolymer poly-3-hydroxybutyrate-co-valerate by Ralstonia eutrophus with organic waste as sole carbon source. J C lean Prod, 1999, 7: 413-420.

[35] Mockos GR, Loge FJ, Smith WA, Thompson DN. Selective enrichment of a methanol- utilizing consortium using pulp & paper mill waste streams. http://www.ncbi.nlm.nih.gov/pubmed/18418753" \o "Applied biochemistry and biotechnology. Appl Biochem Biotechnol, 2008, 148 1–3 211-26.

[36] Rao MN, Datta AK. Waste water treatment, 2007, New Delhi: 702 Co. Pvt. Ltd.

[37] Holt John G, Krieg NR, Peter S, Staley HA, Williams Satnley T, James T. Bergey's manual of determinative bacteriology, 2009, Baltimore, United States: Lippincott Williams & Wilkins.

[38] Mirac Y, Haluk S, Yavuz B. Determination of poly-β-hydroxybutyrate (PHB) production by some Bacillus spp. World J Microbiol Biotechnol, 2004, 21: 565-566.

[39] Quagliano JCF, Amarilla G, Fernandes DM, Miyazaki SS. Effect of simple and complex carbon sources, low temperature culture and complex carbon feeding policies on poly-3-hydroxybutyric acid (PHB) content and molecular weight (Mw) from Azotobacter chroococcum 6B. World J Microbiol Biotechnol, 2001, 17: 9-14.

[40] Pozo C, Martı’nez-Toledo MV, Rodelas B, Gonza’lez-Lo’pez J. Effects of culture conditions on the production of polyhydroxyalkanoates by Azotobacter chroococcum H23 in media containing a high concentration of alpechín (wastewater from olive oil mills) as primary carbon source. J Biotechnol, 2002, 97: 125-131.

[41] Yüksekdağ ZN, Aslim B, Beyatli Y, Mercan N. Effect of carbon and nitrogen sources and incubation times on polybeta-hydroxybutyrate (PHB) synthesis by Bacillus subtilis 25 and Bacillus megaterium 12. African J Biotechnol, 2004, 3 1 63-66.

[42] Khanna S, Srivastava A. Recent advances in microbial polyhydroxyalkanoates. Process Biochem, 2005, 40: 607-619.

[43] Page WJ, Manchak J, Rudy B. Formation of poly(hydroxybutyrate-co-hydroxyvalerate) by Azotobacter vinelandii UWD. Appl Environ Microbiol, 1992, 58: 28-66.

[44] Klu¨ttermann K, Tauchert H, Kleber HP. Synthesis of poly-beta-hydroxybutyrate by Agrobacterium radiobacter after growth on D-Carnitine. Acta Biotechnol, 2002, 22: 261-269.

[45] Singh G, Mittal A, Kumari A, Goyal V, Yadav A, Aggarwal NK. Cost effective production of poly-β-hydroxybutyrate by Bacillus subtilis NG 05 using sugar industry waste water. J Polym Environ, 2013, 21: 441-44.

[46] Tamdoğan N, Sidal U. Investigation of poly-β-Hydroxybutyrate (PHB) production by Bacillus subtilis ATCC 6633 under different conditions. Kafkas Univ Vet Fak Derg, 2011, 17 Suppl A 173-176.

[47] Aslim B, Yüksekdağ ZN, Beyatli Y (2001) Determination of growth quantities of certain Bacillus species isolated from soil. Turk Electr J Biotechnol (Sp. Issue):24–30

[48] Hamieh A, Olama Z, Holail H. Microbial production of polyhydroxybutyrate, a biodegradable plastic using agro-industrial waste products. G Adv Res J Microbiol, 2013, 2 3 054-064.

[49] Sindhu R, Ammu B, Parameswaran B, Deepthi SK, Ramachandran KB, Soccol CR, Pandey A. Improving its thermal properties by blending with other polymers. Brazilian J Microbiol, 2011, 54 4 783-794.

[50] Flora G, Bhatt K, Tuteja U. Optimization of culture conditions for poly-β-hydroxybutyrate production from isolated Bacillus species. J Cell Tissue Res, 2010, 10: 2235-2242.

[51] Shivakumar S. Optimization of process parameters for maximum poly-β-hydroxybutyrate production by Bacillus thuringiensis IAM 12077. Pol J Microbiol, 2009, 58 2 149-154.

[52] Tanamool V, Danvirutai P, Thanonkeo P, Imai T, Kaewkannetra P (2009) Production of poly-β-hydroxybutyric acid (PHB) from sweet sorghum juice by Alcaligenes eutrophus TISTR 1095 and Alcaligenes latus ATCC 29714 via batch fermentation. The 3th International Conference on Fermentation Technology for Value Added Agroculture Products 1–6

[53] El-Sayed AA, Abdelhady HM, Abdel Hafez AM, Khodair TA. Batch production of polyhydroxybutyrate (PHB) by Ralstonia eutropha and Alcaligenes latus using bioreactor different culture strategies. J Appl Sci Res, 2009, 5 5 556-564.

[54] Khanna S, Srivastava A. Optimization of nutrient feed concentration and addition time for production of poly(β-hydroxybutyrate). Enzyme Microbiol Technol, 2006, 39: 1145-1151.

[55] Tanamool V, Imai T, Danvirutai P, Kaewkannetra P. Biosynthesis of polyhydroxyalkanoate (PHA) by Hydrogenophaga sp. isolated from soil environment during batch fermentation. J Life Sci, 2011, 5: 1003-1012.

[56] Yezza A, Halasz A, Levadoux W, Hawari J. Production of poly-β-hydroxybutyrate (PHB) by Alcaligenes latus from maple sap. Apply Microbiol Biotechnol, 2007, 77: 269-274.

[57] Pachekoski WM, Agnelli JAM, Belem LP. Thermal, mechanical and morphological properties of poly (hydroxybutyrate) and polypropylene blends after processing. Material Res, 2009, 12: 159-164.

[58] He W, Tian W, Zhang G, Chen GQ, Zhang Z. Production of novel polyhydroxyalkanoates by Pseudomonas stutzeri 1317 from glucose and soybean oil. FEMS Microbiol Lett, 1998, 169: 45-49.

[59] Galego N, Rozsa C, Sánchez R, Fung J, Vázquez A, Tomás JS. Characterization and application of poly (β-hydroxyalkanoates) family as composite biomaterials. Polym Test, 2000, 19: 485-492.

[60] Choi JY, Lee JK, You Y, Park WH. Epoxidized polybutadiene as a thermal stabilizer for poly(3-hydroxybutyrate). II. Thermal stabilization of poly(3-hydroxybutyrate) by epoxidized polybutadiene. Fiber Polym, 2003, 4: 195-198.

[61] Parra D, Rosa F,D, Rezende SP, Ponce J, Luga˜o AB. Biodegradation of irradiated poly-3-hydroxybutyrate (PHB) films blended with poly (ethyleneglycol). J Polym Environ, 2011, 19: 918-925.