Biosynthesis of Polyhydroxybutyrate from Giant Reed Grass Hydrolysate and Evaluation of its Drug Releasing Profiles
The cost of biopolymer production is one of the most important factor restricting the industrial application of Polyhydroxybutyrate (PHB). The main aim of this present study is to explore suitable low-cost novel substrate for PHB production. In this study, giant reed grass (GRG) biomass was pretreated with different concentration of NaOH (1-3%) at various conditions like room temperature, microwave irradiation high temperature and pressure (HTP). Among these three conditions, at the HTP pretreatment with 2% NaOH, the maximum fermentable glucose of 79.32% was obtained after removal of llignin by SC/AA treatment. Further, this pretreatment condition was used for hydrolysate preparation for PHB biosynthesis using B. subtilis RNM. Under optimized condition, the maximum PHB of 46.21±0.2 gL-1 was obtained from GRG hydrolysate. This result indicates that the GRG hydrolysate could be a novel low-cost substrate for PHB biosynthesis. This is the first attempt we made to use GRG hydrolysate as feedstock for PHB biosynthesis. Moreover, the produced PHB was characterized by 1H NMR, FTIR and thermal amalysis (DSC and TGA) techniques. The characterized results were very similar to characters of commercial PHB. Further, the produced PHB was used for doxorubicin - PHB microparticles preparation with 93.21±0.15 % of encapsulation efficiency and found its anticancer drug delivery efficiency as 98%. Hence, this study demonstrates that the PHB biosynthesized from GRG hydrolysate could be a best biodegradable polymer for anticancer drug doxorubicin encapsulation and delivery.
Keywords: B. subtilis, Biosynthesis, giant reed grass, hydrolysate, microparticles, drug releasing, PHB
2. Gurieff N, Lant P, Comparative life cycle assessment and financial analysis of mixed culture polyhydroxyalkanoates production. Bioresource Technology, 2007; 98:3393-3403.
3. Yu PH, Chua H, Huang AL, Lo W, Chen G-Q, Conversion of food industrial wastes into bioplastics. Applied Biochemistry and Biotechnology, 1998; 70:603-614.
4. Nath A, Dixit M, Bandiya A, Chavda S, Desai AJ, Enhanced PHB production and scale up studies using cheese whey in fed batch culture of Methylobacterium sp. ZP24. Bioresource Technology, 2008; 99: 5749-5755.
5. Naranjo JM, Posada JA, Higuita JC, Cardona CA, Valorization of glycerol through the production of biopolymers: The PHB case using Bacillus megaterium. Bioresource. Technology, 2013;133:38-44
6. Gahlawat G, Soni SK, Valorization of waste glycerol for the production of poly (3-hydroxybutyrate) and poly (3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer by Cupriavidus necator and extraction in a sustainable manner, Bioresource Technology, 2017; 243: 492-501.
7. Foust TD, Aden A, Dutta A, Phillips S, An economic and environmental comparison of a biochemical and a thermochemical lignocellulosic ethanol conversion processes, Cellulose, 2006; 16:547-565.
8. Zhang T, Kumar R, Wyman CE,Sugar yields from dilute oxalic acid pre-treatment of maple wood compared to those with other dilute acids and hot water, Carbohydrate Polymers, 2013; 92(1): 334-344.
9. Santos JC, Marton JM, Felipe MGA, Continuous system of combined columns of ion exchange resins and activated charcoal as a new approach for the removal of toxics from sugar cane bagasse hemicellulosic hydrolysate, Industrial & Engineering Chemistry Research, 2014; 53(42): 16494-16501.
10. Mtui GYS, Oxalic acid pretreatment, fungal enzymatic saccharification and fermentation of maize residues to ethanol, African Journal of Biotechnology, 2012; 11(4): 843-851.
11. Scordia D, Cosentino SL, Jeffries TW, Effectiveness of dilute oxalic acid pre-treatment of Miscanthus xgiganteus biomass for ethanol production, Biomass Bioenergy, 2013; 59: 540-548.
12. Davis R, Kataria R, Cerrone F, Woods T, Kenny S, O’Donovan A, Guzik M, Shaikh H, Duane G, Gupta VK, Tuohy MG, Padamatti RB, Casey E, O’Connor KE, Conversion of grass biomass into fermentable sugars and its utilization for medium chain length polyhydroxyalkanoate (mcl-PHA) production by Pseudomonas strains, Bioresource Technology, 2013; 150: 202-209.
13. Han JS, Rowell JS, Chemical composition of fibers. In: Rowell, R.M., Young, R.A., Rowell, J.K. (Eds.), Chemical Composition of Fibers, Paper Composites from Agro-based Resources. CRC Lewis Publisher, New York, 1997; pp. 83–134.
14. Borman AM, Szekely A, Campbell CK, Johnson EM, Evaluation of the viability of pathogenic filamentous fungi after prolonged storage in sterile water and review of recent published studies on storage methods. Mycopathologia, 2006;, 161: 361-368.
15. Miller GL, Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 1959; 31: 426-428.
16. Li H, Long C, Zhou J, Liu J, Wu X, Long M, Rapid analysis of mono-saccharides and oligo-saccharides in hydrolysates of lignocellulosic biomass by HPLC, Biotechnology Letter, 2013; 35:1405–1409.
17. Zhang LY, Xu J, Zhang LH, Zhang WB, Zhang YK, Determination of 1-phenyl-3-methyl-5-pyrazolone-labeled carbohydrates by liquid chromatography and micellar electrokinetic chromatography, Journal of Chromatogr B, 2003;793:159–165.
18. Koller M, Bona R, Hermann C, Horvat P, Martinz J, Neto J, Varila P, Braunegg G, Biotechnological production of poly (3-hydroxybutyrate) with Wautersia eutropha by application of green grass juice and silage juice as additional complex substrates. Biocatal. Biotransform, 2005; 23: 329-337.
19. Sayyed RZ, Gangurde NS, Chincholkar SB, Hypochlorite digestion method for efficient recovery of PHB from Alcaligenes faecalis. Indian Journal of Microbiology, 2009; 49: 230-232.
20. Law JH, Slepecky RA, Assay of poly-β-hydroxybutric acid. Journal of Bacteriology, 1961; 82(1): 33-36.
21. Kato M, Bao HJ, Kang CK, Fukui T, Doi Y, Production of a novel copolyester of 3-hydroxybutyric acid and medium-chain-length 3- hydroxyalkanoic acids by Pseudomonas sp. 61-3 from sugars, Applied Microbiology and Biotechnology, 1996; 45:363-370.
22. Bazzo GC, Lemos-Senna E, Goncalves MC, Pires ATN, Effect of preparation conditions on morphology, drug content and release profiles of poly(hydroxybutyrate) microparticles containing piroxicam, Journal of the Brazilian Chemical Society, 2008; 19( 5): 914-921.
23. Taghavi S, Garafola C, Monchy S, Lee M, Hoffman A, Weyens N, Barac T, Jaco V, van der Lelie D, Genome survey and characterization of entophytic bacteria exhibiting a beneficial effect on growth and development of poplar trees. Applied Environmental Microbiology, 2009; 75: 748–757.
24. Le Meur S, Zinn M, Egli T, Thony-Meye, L, Ren Q, Production of mediumchain-length polyhydroxyalkanoates by sequential feeding of xylose and octanoic acid in engineered Pseudomonas putida KT2440. BMC Biotechnology, 2012; 12:53.
25. Kulpreecha S, Boonruangthavorn A, Meksiriporn B, Thongchul N, Inexpensive fed-batch cultivation for high Poly (3-Hydroxybutyrate) production by a new isolate Bacillus megaterium. Journal of Bioscience and Bioengineering, 2009; 107: 240-245.
26. Ma L, Zhang H, Liu Q, Chen J, Zhang J, Chen GQ, Production of two monomer structures containing medium-chain-length polyhydroxyalkanoates by β-oxidation-impaired mutant of Pseudomonas putida KT2442. Bioresource Technology, 2009; 100:4891-4894.
27. Vishnuvardhan RS, Thirumala M, Mahmood S, Production of PHB and P (3HB-co-3HV) biopolymers by Bacillus megaterium strain OU303A isolated from municipal sewage sludge, World Journal of Microbiology and Biotechnology, 2009; 25:391-397.
28. Hahn SK, Chang YK, Kim BS, Chang HN, Optimization of microbial Poly (3hydroxybutyric Acid) recovery using dispersions of sodium hypochlorite solution and chloroform. Biotechnology and Bioengineering, 1994; 44:256-261.
29. Sudesh K, Abe H, Doi Y, Structure and properties of polyhydroxyalkanoate, biological polyesters. Progress in Polymer Science, 2000; 25(10): 1503-1533.
30. Williams S, Martin D, Applications of Polyhydroxyalkanoate (PHA) in medicine and pharmacy, Biopolymers, 2005; 91-103.
31. Tripathi AD, Joshi TR, Srivastava SK, Darani KK, Shankar Khade S, Srivastava J, Effect of nutritional supplements on bioplastics (PHB) production utilizing sugar refinery waste with potential application in food packaging, Preparative Biochemistry Biotechnology, 2019; 49(6):567-577.
32. Chaijamrus S, Udpuay N, Production and characterization of Polyhydroxybutyrate from molasses and corn steep liquor produced by Bacillus megaterium ATCC 6748, Agricultural Engineering International: the CIGR E. journal. Manuscript FP, 2008; 10:1-12.
33. Astra LI, Hammond R, Tarakji K, Stephenson LW, Doxourbibin-induced canine CHF: advantages and disadvantages, Journal of Cardiac Surgery, 2003; 18(4): 301-306.
34. Fresta M, Puglisi G, Giammona G, Cavallaro G, Micali N, Furneri PM, Pefloxacine mesilate- and ofloxacin-loaded polyethylcyanoacrylate nanoparticles: characterization of the colloidal drug carrier formulation, Journal of Pharmaceutical Science, 1995; 84 (7): 895- 902.
35. Govender T, Riley T, Ehtezazi T, Garnett MC, Stolnik S, Illum L, Davis SS, Defining the drug incorporation properties of PLA–PEG nanoparticles, International Journal of Pharmaceutics, 2000; 199 (1):95–110.
36. Panyam J, Williams D, Dash A, Leslie-Pelecky D, Labhasetwar V, Solid-state solubility influences encapsulation and release of hydrophobic drugs from PLGA/PLA nanoparticles, Journal of Pharmaceutical Science, 2004; 93 (7): 1804–1814.
37. Kumari A, Yadav SK, Pakade YB, Singh B, Yadava SC, Development of biodegradable nanoparticles for delivery of quercetin. Colloids Surf B Biointerfaces, 2010; 80: 184–192.
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