Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Novel Polyhydroxybutyrate-Degrading Activity of the Microbulbifer Genus as Confirmed by Microbulbifer sp. SOL03 from the Marine Environment

  • Park, Sol Lee (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Cho, Jang Yeon (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Kim, Su Hyun (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Lee, Hong-Ju (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Kim, Sang Hyun (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Suh, Min Ju (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Ham, Sion (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Bhatia, Shashi Kant (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Gurav, Ranjit (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Park, ee-Hyoung (Department of Biological and Chemical Engineering, Hongik University) ;
  • Park, Kyungmoon (Department of Biological and Chemical Engineering, Hongik University) ;
  • Kim, Yun-Gon (Department of Chemical Engineering, Soongsil University) ;
  • Yang, Yung-Hun (Department of Biological Engineering, College of Engineering, Konkuk University)
  • Received : 2021.09.03
  • Accepted : 2021.10.25
  • Published : 2022.01.28
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Abstract

Ever since bioplastics were globally introduced to a wide range of industries, the disposal of used products made with bioplastics has become an issue inseparable from their application. Unlike petroleum-based plastics, bioplastics can be completely decomposed into water and carbon dioxide by microorganisms in a relatively short time, which is an advantage. However, there is little information on the specific degraders and accelerating factors for biodegradation. To elucidate a new strain for biodegrading poly-3-hydroxybutyrate (PHB), we screened out one PHB-degrading bacterium, Microbulbifer sp. SOL03, which is the first reported strain from the Microbulbifer genus to show PHB degradation activity, although Microbulbifer species are known to be complex carbohydrate degraders found in high-salt environments. In this study, we evaluated its biodegradability using solid- and liquid-based methods in addition to examining the changes in physical properties throughout the biodegradation process. Furthermore, we established the optimal conditions for biodegradation with respect to temperature, salt concentration, and additional carbon and nitrogen sources; accordingly, a temperature of 37℃ with the addition of 3% NaCl without additional carbon sources, was determined to be optimal. In summary, we found that Microbulbifer sp. SOL03 showed a PHB degradation yield of almost 97% after 10 days. To the best of our knowledge, this is the first study to investigate the potent bioplastic degradation activity of Microbulbifer sp., and we believe that it can contribute to the development of bioplastics from application to disposal.

Keywords

Acknowledgement

This paper was supported by Konkuk University Researcher Fund in 2021. This study also was supported by the National Research Foundation of Korea (NRF) (NRF-2019R1F1A1058805 and NRF-2019M3E6A1103979) and by the R&D Program of MOTIE/KEIT (20009508 and 20016324). This research was also supported by "Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ0154982021), Rural Development Administration, Republic of Korea.

References

  1. Emadian SM, Onay TT, Demirel B. 2017. Biodegradation of bioplastics in natural environments. Waste Manag. 59: 526-536. https://doi.org/10.1016/j.wasman.2016.10.006
  2. Romen F, Reinhardt S, Jendrossek D. 2004. Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegellella sp. KB1a. Arch. Microbiol. 182: 157-164.
  3. Jendrossek D, Knoke I, Habibian RB, Steinbuchel A, Schlegel HG. 1993. Degradation of poly(3-hydroxybutyrate), PHB, by bacteria and purification of a novel PHB depolymerase from Comamonas sp. J. Environ.Polym. Degrad. 1: 53-63. https://doi.org/10.1007/BF01457653
  4. Mukai K, Yamada K, Doi Y. 1993. Kinetics and mechanism of heterogeneous hydrolysis of poly[(R)-3-hydroxybutyrate] film by PHA depolymerases. Int. J. Biol. Macromol. 15: 361-366. https://doi.org/10.1016/0141-8130(93)90054-P
  5. Papaneophytou CP, Pantazaki AA, Kyriakidis DA. 2009. An extracellular polyhydroxybutyrate depolymerase in Thermus thermophilus HB8. Appl. Microbiol. Biotechnol. 83: 659-668. https://doi.org/10.1007/s00253-008-1842-2
  6. Jendrossek D, Schirmer A, Schlegel HG. 1996. Biodegradation of polyhydroxyalkanoic acids. Appl. Mcrobiol. Biotechnol. 46: 451-463. https://doi.org/10.1007/s002530050844
  7. Iwata T, Doi Y, Tanaka T, Akehata T, Shiromo M, Teramachi S. 1997. Enzymatic degradation and adsorption on poly[(R)-3-hydroxybutyrate] single crystals with two types of extracellular PHB depolymerases from Comamonas acidovorans YM1609 and Alcaligenes faecalis T1. Macromolecules 30: 5290. https://doi.org/10.1021/ma970491g
  8. Kasuya K, Inoue Y, Yamada K, Doi Y. 1995. Kinetics of surface hydrolysis of poly[(R)-3-hydroxybutyrate] film by PHB depolymerase from Alcaligenes faecalis T1. Polym. Degrad. Stability 48: 167-174. https://doi.org/10.1016/0141-3910(95)00026-I
  9. Gonzalez JM, Mayer F, Moran MA, Hodson RE, Whitman WB. 1997. Microbulbifer hydrolyticus gen. nov., sp. nov., and Marinobacterium georgiense gen. nov., sp. nov., two marine bacteria from a lignin-rich pulp mill waste enrichment community. Int. J. Syst. Bacteriol 47: 369-376. https://doi.org/10.1099/00207713-47-2-369
  10. Nishijima M, Takadera T, Imamura N, Kasai H, An KD, Adachi K, et al. 2009. Microbulbifer variabilis sp. nov. and Microbulbifer epialgicus sp. nov., isolated from Pacific marine algae, possess a rod-coccus cell cycle in association with the growth phase. Int. J. Syst. Evol. Microbiol. 59: 1696-1707. https://doi.org/10.1099/ijs.0.006452-0
  11. Ritzmann NH, Mahrlein A, Ernst S, Hennecke U, Drees SL, Fetzner S. 2019. Bromination of alkyl quinolones by Microbulbifer sp. HZ11, a marine Gammaproteobacterium, modulates their antibacterial activity. Environ. Microbiol. 21: 2595-2609. https://doi.org/10.1111/1462-2920.14654
  12. Gurav R, Bhatia SK, Choi TR, Jung HR, Yang SY, Song HS, et al. 2019. Chitin biomass powered microbial fuel cell for electricity production using halophilic Bacillus circulans BBL03 isolated from sea salt harvesting area. Bioelectrochemistry 130: 107329. https://doi.org/10.1016/j.bioelechem.2019.107329
  13. Park YL, Bhatia SK, Gurav R, Choi TR, Kim HJ, Song HS, et al. 2020. Fructose based hyper production of poly-3-hydroxybutyrate from Halomonas sp. YLGW01 and impact of carbon sources on bacteria morphologies. Int. J. Biol. Macromol. 154: 929-936. https://doi.org/10.1016/j.ijbiomac.2020.03.129
  14. Uchida H, Nakajima-Kambe T, Shigeno-Akutsu Y, Nomura N, Tokiwa Y, Nakahara T. 2000. Properties of a bacterium which degrades solid poly(tetramethylene succinate)-co-adipate, a biodegradable plastic. FEMS Microbiol. Lett. 189: 25-29. https://doi.org/10.1016/S0378-1097(00)00246-9
  15. Park SL, Cho JY, Choi TR, Song HS, Bhatia SK, Gurav R, et al. 2021. Improvement of polyhydroxybutyrate (PHB) plate-based screening method for PHB degrading bacteria using cell-grown amorphous PHB and recovered by sodium dodecyl sulfate (SDS). Int. J. Biol. Macromol. 177: 413-421. https://doi.org/10.1016/j.ijbiomac.2021.02.098
  16. Bhatia SK, Yoon JJ, Kim HJ, Hong JW, Hong YG, Song HS, et al. 2018. Engineering of artificial microbial consortia of Ralstonia eutropha and Bacillus subtilis for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer production from sugarcane sugar without precursor feeding. Bioresour. Technol. 257: 92-101. https://doi.org/10.1016/j.biortech.2018.02.056
  17. Jung HR, Lee JH, Moon YM, Choi TR, Yang SY, Song HS, et al. 2019. Increased tolerance to furfural by introduction of polyhydroxybutyrate synthetic genes to Escherichia coli. J. Microbiol. Biotechnol. 29: 776-784. https://doi.org/10.4014/jmb.1901.01070
  18. Jung HR, Yang SY, Moon YM, Choi TR, Song HS, Bhatia SK, et al. 2019. Construction of efficient platform Escherichia coli strains for polyhydroxyalkanoate production by engineering branched pathway. Polymers 11: 509. https://doi.org/10.3390/polym11030509
  19. Braunegg G, Sonnleitner B, Lafferty RM. 1978. A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur. J. Appl. Microbiol. Biotechnol. 6: 29-37. https://doi.org/10.1007/BF00500854
  20. Hong YG, Moon YM, Hong JW, Choi TR, Jung HR, Yang SY, Jang DW, et al. 2019. Discarded egg yolk as an alternate source of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). J. Microbiol. Biotechnol. 29: 382-391. https://doi.org/10.4014/jmb.1811.11028
  21. Ham S, Han YH, Kim SH, Suh MJ, Cho JY, Lee HJ, et al. 2021. Application of l-glutamate oxidase from Streptomyces sp. X119-6 with catalase (KatE) to whole-cell systems for glutaric acid production in Escherichia coli. Kor. J. Chem. Eng. 38: 2106-2112. https://doi.org/10.1007/s11814-021-0855-8
  22. Jung HR, Choi TR, Han YH, Park YL, Park JY, Song HS, et al. 2020. Production of blue-colored polyhydroxybutyrate (PHB) by one-pot production and coextraction of indigo and PHB from recombinant Escherichia coli. Dyes Pigm. 173: 107889-107896. https://doi.org/10.1016/j.dyepig.2019.107889
  23. Gurav R, Bhatia SK, Choi TR, Park YL, Park JY, Han YH, et al. 2020. Treatment of furazolidone contaminated water using banana pseudostem biochar engineered with facile synthesized magnetic nanocomposites. Bioresour. Technol. 297: 122472. https://doi.org/10.1016/j.biortech.2019.122472
  24. Bhatia SK , Gurav R, Choi TR, Jung HR, Yang SY, Song HS, et al. 2019. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) production from engineered Ralstonia eutropha using synthetic and anaerobically digested food waste derived volatile fatty acids. Int. J. Biol. Macromol. 133: 1-10. https://doi.org/10.1016/j.ijbiomac.2019.04.083
  25. Kuchta K, Chi L, Fuchs H, Potter M, Steinbuchel A. 2007. Studies on the influence of phasins on accumulation and degradation of PHB and nanostructure of PHB granules in Raistonia eutropha H16. Biomacromolecules 8: 657-662. https://doi.org/10.1021/bm060912e
  26. Choi TR, Jeon JM, Bhatia SK, Gurav R, Han YH, Park YL, et al. 2020. Production of low molecular weight P(3HB-co-3HV) by butyrateacetoacetate CoA-transferase (cftAB) in Escherichia coli. Biotechnol. Bioprocess Eng. 25: 279-286. https://doi.org/10.1007/s12257-019-0366-1
  27. Pattanasuttichonlakul W, Sombatsompop N, Prapagdee B. 2018. Accelerating biodegradation of PLA using microbial consortium from dairy wastewater sludge combined with PLA-degrading bacterium. Int. Biodeterior. Biod. 132: 74-83. https://doi.org/10.1016/j.ibiod.2018.05.014
  28. Kampfer P, Arun AB, Young CC, Rekha PD, Martin K, Busse HJ. et al. 2012. Microbulbifer taiwanensis sp. nov., isolated from coastal soil. Int. J. Syst. Evol. Microbiol. 62: 2485-2489. https://doi.org/10.1099/ijs.0.034512-0
  29. Yoon JH, Kim IG, Oh TK, Park YH. 2004. Microbulbifer maritimus sp. nov., isolated from an intertidal sediment from the Yellow Sea, Korea. Int. J. Syst. Evol. Microbiol. 54: 1111-1116. https://doi.org/10.1099/ijs.0.02985-0
  30. Lodhi AF, Hasan F, Shah Z, Hameed A, Faisal S, Shah AA. 2011. Optimization of culture conditions for the production of poly (3-hydroxybutyrate) depolymerase from newly isolated Aspergillus fumigatus from soil. Pak. J. Bot. 43: 1361-1372.
  31. Manna A, Paul AK. 2000. Degradation of microbial polyester poly(3-hydroxybutyrate) in environmental samples and in culture. Biodegradation 11: 323-339 . https://doi.org/10.1023/A:1011162624704
  32. Vigneswari S, Lee TS, Bhubalan K, Amirul AA. 2015. Extracellular Polyhydroxyalkanoate depolymerase by Acidovorax sp. DP5. Enzyme Res. 2015: 212159. https://doi.org/10.1155/2015/212159
  33. Iwata T, Doi Y, Kasuya KI, Inoue Y. 1997. Visualization of enzymatic degradation of poly[(R)-3-hydroxybutyrate] single crystals by an extracellular PHB depolymerase. Macromolecules 30: 833-839. https://doi.org/10.1021/ma961352m
  34. Kasuya KI, Inoue Y, Doi Y. 1996. Adsorption kinetics of bacterial PHB depolymerase on the surface of polyhydroxyalkanoate films. Int. J. Biol. Macromol. 19: 35-40. https://doi.org/10.1016/0141-8130(96)01097-5
  35. JX and JP Spallas. 2012. Different contrast mechanisms in SEM imaging of graphene. Agilent Technologies.
  36. Sridewi N, Bhubalan K, Sudesh K. 2006. Degradation of commercially important polyhydroxyalkanoates in tropical mangrove ecosystem. Polymer Degrad. Stab. 91: 2931-2940. https://doi.org/10.1016/j.polymdegradstab.2006.08.027
  37. Wang YW, Mo W, Yao H, Wu Q, Chen J, Chen GQ. 2004. Biodegradation studies of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). Polym. Degrad. Stab. 85: 815-821. https://doi.org/10.1016/j.polymdegradstab.2004.02.010
  38. Won NI, Lee GE, Ko K, Oh DC, Na YH, Park JS. 2017. Identification of a bioactive compound, violacein, from Microbulbifer sp. isolated from a marine sponge Hymeniacidon sinapium on the west coast of Korea. Microbiol. Biotechnol. Lett. 45: 124-132. https://doi.org/10.4014/mbl.1702.02002
  39. Huang H, Mo K, Hu Y, Liu M, Zhu J, Zou X, et al. 2020. Microbulbifer arenosus sp. nov., an alginate-degrading bacterium isolated from coastal sand. Int. J. Syst. Evol. Microbiol. 70: 1639-1643. https://doi.org/10.1099/ijsem.0.003945
  40. Vashist P, Nogi Y, Ghadi SC, Verma P, Shouche YS. 2013. Microbulbifer mangrovi sp. nov., a polysaccharide-degrading bacterium isolated from an Indian mangrove. Int. J. Syst. Evol. Microbiol. 63: 2535-2537.
  41. Sun C, Chen YJ, Zhang XQ, Pan J, Cheng H, Wu M. 2014. Draft genome sequence of Microbulbifer elongatus strain HZ11, a brown seaweed-degrading bacterium with potential ability to produce bioethanol from alginate. Mar. Genomics 18pt B: 83-85. https://doi.org/10.1016/j.margen.2014.05.009
  42. Tanaka D, Ohnishi KI, Watanabe S, Suzuki S. 2021. Isolation of cellulase-producing microbulbifer sp. from marine teleost blackfish (Girella melanichthys) intestine and the enzyme characterization. J. Gen. Appl. Microbiol. 67: 47-53. https://doi.org/10.2323/jgam.2020.05.001
  43. Takagi E, Hatada Y, Akita M, Ohta Y, Yokoi G, Miyazaki T. 2015. Crystal structure of the catalytic domain of a GH16 β-agarase from a deep-sea bacterium, Microbulbifer thermotolerans JAMB-A94. Biosci. Biotechnol. Biochem. 79: 625-632. https://doi.org/10.1080/09168451.2014.988680
  44. Miyazaki M, Nogi Y, Ohta Y, Hatada Y, Fujiwara Y, Ito S. 2008. Microbulbifer agarilyticus sp. nov. and Microbulbifer thermotolerans sp. nov., agar-degrading bacteria isolated from deep-sea sediment. Int. J. Syst. Evol. Microbiol. 58: 1128-1133. https://doi.org/10.1099/ijs.0.65507-0