Polyhydroxyalkanoates (PHAs) are polymers with biodegradable and biocompatible properties accumulated in a wide variety of bacterial strains. In the present study, active sludge, wheat starch wastewater (WSW), and oil wastewater were used for the isolation and screening of PHA-accumulating bacteria. WSW was then implemented as a cheap and economical culture medium for the production of PHAs by the selected isolate. The extracted PHA was characterized, and the capability of produced biopolymer for preparing nanoparticles was evaluated. Based on the results, 96 different bacterial isolates were obtained, of which the strains isolated from WSW demonstrated the highest PHA-accumulation capability. The maximum PHA content of 3.07 g/l (59.50% of dry cell weight) was obtained by strain N6 in 21 h. The selected strain was identified by molecular approaches as Bacillus cereus. Afterward, the physicochemical characterization of an accumulated biopolymer was specified as a PHBV copolymer. Finally, spherical homogenous PHBV nanoparticles with a size of 137 nm were achieved. The PHBV nanoparticles showed a suitable small size and good zeta potential for medical applications. Hence, it can be concluded that isolated wild strain (B. cereus) has the potential exploitation capability for cost-effective PHBV production using the WSW.
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All isolated strains are preserved in Persian Type Culture Collection (PTCC) and data are available by N. Sinaei.
Luengo JM, Garcia B, Sandoval A, Naharro G, Olivera ER (2003) Bioplastics from microorganisms. Curr Opin Microbiol 6:251–260. https://doi.org/10.1016/s1369-5274(03)00040-7
Akaraonye E, Keshavarz T, Roy I (2010) Production of polyhydroxyalkanoates: the future green materials of choice. J Chem Technol Biotechnol 85:732–743. https://doi.org/10.1002/jctb.2392
Peña C, López S, García A, Espín G, Romo-Uribe A, Segura D (2014) Biosynthesis of poly-βhydroxybutyrate (PHB) with a high molecular mass by a mutant strain of Azotobacter vinelandii (OPN). Ann Microbiol 64:39–47. https://doi.org/10.1007/s13213-013-0630-0
Batcha AFM, Prasad DMR, Khan MR, Abdullah H (2014) Biosynthesis of poly(3- hydroxybutyrate) (PHB) by Cupriavidus necator H16 from jatropha oil as carbon source. Bioprocess Biosyst Eng 37:943–951. https://doi.org/10.1007/s00449-013-1066-4
Song R, Murphy M, Li C, Ting K, Soo C, Zheng Z (2018) Current development of biodegradable polymeric materials for biomedical applications. Drug Des Dev Ther 12:3117–3145. https://doi.org/10.2147/DDDT.S165440
Mohan S, Oluwafemi OS, Kalarikkal N, Thomas S, Songca SP (2016) Biopolymers– application in nanoscience and nanotechnology. Recent Adv Biopolymers 47. https://doi.org/10.5772/62225
Ozdil D, Aydin HM (2014) Polymers for medical and tissue engineering applications. J Chem Technol Biotechnol 89(12):1793–1810. https://doi.org/10.1002/jctb.4505
Philip S, Keshavarz T, Roy I (2007) Polyhydroxyalkanoates: biodegradable polymers with a range of applications. J Chem Technol Biotechnol 82:233–247. https://doi.org/10.1002/jctb.1667
Kerketta A, Vasanth D (2019) Madhuca indica flower extract as cheaper carbon source for production of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) using Ralstonia eutropha. Process Biochem 87:1–9. https://doi.org/10.1016/j.procbio.2019.09.013
Chen GQ, Wu Q (2005) The application of polyhydroxyalkanoates as tissue engineering materials. Biomaterials 26(33):6565–6578. https://doi.org/10.1016/j.biomaterials.2005.04.036
Rivera-Briso AL, Serrano-Aroca Á (2018) Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate): enhancement strategies for advanced applications. Polymers 10(7):732–759
Halami PM (2008) Production of polyhydroxyalkanoate from starch by the native isolate Bacillus cereus CFR06. World J Microbiol Biotechnol 24:805–812. https://doi.org/10.1007/s11274-007-9543-z
Sharma P, Bajaj B (2015) Cost-effective substrates for production of poly-β-hydroxybutyrate by a newly isolated Bacillus cereus PS-10. J Environ Biol 36:1297–1304
Naheed N, Jamil N (2014) Optimization of biodegradable plastic production on sugar cane molasses in Enterobacter sp. SEL2. Braz J Microbiol 45:417–426. https://doi.org/10.1590/s1517-83822014000200008
Bhattacharyya A, Pramanik A, Maji SK, Haldar S, Mukhopadhyay UK, Mukherjee J (2012) Utilization of vinasse for production of poly-3-(hydroxybutyrate-co-hydroxyvalerate) by Haloferax mediterranei. AMB Express 9:2(1):34. https://doi.org/10.1186/2191-0855-2-34
Sathiyanarayanan G, Kiran GS, Selvinc J, Saibaba G (2013) Optimization of polyhydroxybutyrate production by marine Bacillus megaterium MSBN04 under solid state culture. Int J Biol Macromol 60:253–261. https://doi.org/10.1016/j.ijbiomac.2013.05.031
Haas R, Jin B, Zepf FT (2008) Production of poly(3-hydroxybutyrate) from waste potato starch. Biosci Biotechnol Biochem 72:253–256. https://doi.org/10.1271/bbb.70503
Dahman Y, Ugwu CU (2014) Production of green biodegradable plastics of poly(3-hydroxybutyrate) from renewable resources of agricultural residues. Bioprocess Biosyst Eng 37:1561–1568. https://doi.org/10.1007/s00449-014-1128-2
Novackova I, Kucera D, Porizka J, Pernicova I, Sedlacek P, Koller M, Kovalcik A, Obruca S (2019) Adaptation of Cupriavidus necator to levulinic acid for enhanced production of P(3HB-co-3HV) copolyesters. Biochem Eng J 151:107350. https://doi.org/10.1016/j.bej.2019.107350
Farrag Y, Montero B, Rico M, Barral L, Bouza R (2018) Preparation and characterization of nano and micro particles of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) via emulsification/solvent evaporation and nanoprecipitation techniques. J Nanopart Res 20:71. https://doi.org/10.1007/s11051-018-4177-7
Wei YH, Chen WC, Huang CK, Wu HS, Sun YM, Lo CW, Janarthanan OM (2011) Screening and evaluation of Polyhydroxybutyrate-producing strains from indigenous isolate Cupriavidus taiwanensis strains. Int J Mol Sci 12:252–265. https://doi.org/10.3390/ijms12010252
Miller GL (1959) Use of dinitrosaIicyIic acid reagent for determination of reducing sugar. Anal Chem 31:426–428. https://doi.org/10.1021/ac60147a030
Singh G, Kumari A, Mittal A, Yadav A, Aggarwal NK (2013) Poly β-Hydroxybutyrate production by Bacillus subtilis NG220 using sugar industry waste water. BioMed Research International Article ID 952641. https://doi.org/10.1155/2013/952641
Law J, Slepecky RA (1961) Assay of poly-beta-hydroxybutyric acid. J Bacteriol 82(1):33–36. https://doi.org/10.1128/JB.82.1.33-36.1961
Hahn SK, Chang YK, Kim BS, Lee KM, Chang HN (1993) The recovery of poly(3-hydroxybutyrate) by using dispersions of sodium hypochlorite solution and chloroform. Biotechnol Tech 7(3):209–212. https://doi.org/10.1007/BF02566149
Shrivastava A, Mishraa SK, Shethia B, Pancha I, Jain D, Mishraa S (2010) Isolation of promising bacterial strains from soil and marine environment for polyhydroxyalkanoates (PHAs) production utilizing Jatropha biodiesel byproduct. Int J Biol Macromol 47:283–287. https://doi.org/10.1016/j.ijbiomac.2010.04.007
Preethi R, Sasikala P, Aravind J (2012) Microbial production of polyhydroxyalkanoate (PHAs) utilizing fruit waste as a substrate. Res in Biotech 3(1):61–69
Contreras AR, Koller M, Dias MMS, Monfort MC, Braunegg G, Calvo MSM (2013) High production of poly (3-hydroxybutyrate) from a wild Bacillus megaterium bolivian strain. J Appl Microbiol 114(5):1378–1387. https://doi.org/10.1111/jam.12151
Öner M, Çöl AA, Pochat-Bohatier C, Bechelany M (2016) Effect of incorporation of boron nitride nanoparticles on the oxygen barrier and thermalproperties of poly(3-hydroxybutyrate-cohydroxyvalerate). RSC Adv 6:90973–90981. https://doi.org/10.1039/C6RA19198C
Vilos C, Morales FA, Solar PA, Herrera NS, Gonzalez-Nilo FD, Aguayo DA, Mendoza HL, Comer J, Bravo ML, Gonzalez PA, Kato S, Cuello MA, Alonso C, Bravo EJ, Bustamante EI, Owen GI, Velasquez LA (2013) Paclitaxel-PHBV nanoparticles and their toxicity to endometrial and primary ovarian cancer cells. Biomaterials 34(16):4098–4108. https://doi.org/10.1016/j.biomaterials.2013.02.034
Liu H, Pancholi M, Stubbs J III, Raghavan D (2010) Influence of hydroxyvalerate composition of polyhydroxy butyrate valerate (PHBV) copolymer on bone cell viability and in vitro degradation. J Appl Polym Sci 116:3225–3231. https://doi.org/10.1002/app.31915
Bayar S, Severcan F (2005) FTIR study of biodegradable biopolymers: P(3HB), P(3HB-co-4HB) and P(3HB-co-3HV). J Mol Struct 744:529–534. https://doi.org/10.1016/j.molstruc.2004.12.029
Moorkoth D, Nampoothiri KM (2016) Production and characterization of poly(3-hydroxy butyrate-co-3hydroxyvalerate) (PHBV) by a novel halotolerant mangrove isolate. Bioresour Technol 201:253–260. https://doi.org/10.1016/j.biortech.2015.11.046
Mohapatra S, Mohanta PR, Sarkar B, Daware A, Kumar C, Samantaray DP (2015) Production of Polyhydroxyalkanoates (PHAs) by Bacillus strain isolated from waste water and its biochemical characterization. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 87(2):459–466. https://doi.org/10.1007/s40011-015-0626-6
Bhuwal AK, Singh G, Aggarwal NK, Goyal V, Yadav A (2014) Poly-β-hydroxybutyrate production and management of cardboard industry effluent by new Bacillus sp. NA10. Bioresour. Bioprocess. 1, 9. https://doi.org/10.1186/s40643-014-0009-5
Aslim B, Yüksekdağ ZN, Beyatli Y (2002) Determination of PHB growth quantities of certain bacillus species isolated from soil. Turk.E.J. Biotechnol. 2:24–30
Labuzek S, Radecka I (2001) Biosynthesis of PHB tercopolymer by Bacillus cereus UW85. J Appl Microbiol 90:353–357. https://doi.org/10.1046/j.1365-2672.2001.01253.x
Yilmaz M, Soran H, Beyatli Y (2005) Determination of poly-b-hydroxybutyrate (PHB) production by some Bacillus spp. World J Microbiol Biotechnol 21:565–566. https://doi.org/10.1007/s11274-004-3274-1
Tufail S, Munir S, Jamil N (2017) Variation analysis of bacterial polyhydroxyalkanoates production using saturated and unsaturated hydrocarbons. Braz J Microbiol 48(4):629–636. https://doi.org/10.1016/j.bjm.2017.02.008
Liu Z, Wang Y, He N, Huang J, Zhu K, Shao W, Wang H, Yuan W, Li Q (2011) Optimization of polyhydroxybutyrate (PHB) production by excess activated sludge and microbial community analysis. J Hazard Mater 185:8–16. https://doi.org/10.1016/j.jhazmat.2010.08.003
Masood F, Hasan F, Ahmed S, Chen P, Hameed A (2012) Biosynthesis and characterization of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) from Bacillus cereus S10. J Polym Environ 20:865–871. https://doi.org/10.1007/s10924-012-0457-y
Rivera-Briso AL, Aachmann FL, Moreno-Manzano V, Serrano-Aroca Á (2020) Graphene oxide nanosheets versus carbon nanofibers: enhancement of physical and biological properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) films for biomedical applications. Int J Biol Macromol 143:1000–1008. https://doi.org/10.1016/j.ijbiomac.2019.10.034
Lombardo D, Kiselev MA, Caccamo MT (2019) Smart nanoparticles for drug delivery application: development of versatile nanocarrier platforms in biotechnology and nanomedicine. J Nanomater 2019:1–26. https://doi.org/10.1155/2019/3702518
Decuzzi P, Ferrari M (2007) The role of specific and non-specific interactions in receptor-mediated endocytosis of nanoparticles. Biomaterials 28:2915–2922. https://doi.org/10.1016/j.biomaterials.2007.02.013
The authors are thankful to the Commonwealth Scientific and Industrial Research Organization (CSIRO), Australia, for providing laboratory facilities for nanoparticle study. We are also grateful to Ms. Mohseni, the head of the Persian Type Culture Collection (PTCC), for identifying the isolated strains.
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Sinaei, N., Zare, D. & Azin, M. Production and characterization of poly 3-hydroxybutyrate-co-3-hydroxyvalerate in wheat starch wastewater and its potential for nanoparticle synthesis. Braz J Microbiol (2021). https://doi.org/10.1007/s42770-021-00430-5
- Poly (3-hydroxybutyrate-co-3-hydroxyvalerate)
- Wheat starch wastewater
- Bacillus cereus
- Medical application