Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Pseudomonas sp. AOB-7 utilizes PHA granules as a sustained-release carbon source and biofilm carrier for aerobic denitrification of aquaculture water

Abstract

Nitrate accumulation causes long-time threat to aquatic animals in recirculating aquaculture system (RAS); thus, nitrate removal is also required in RASs. However, the lack of carbon sources makes denitrification difficult to function. Nitrate removal performance of an aerobic denitrifying and extracellular polyhydroxyalkanoate depolymerase–producing bacterium, Pseudomonas sp. AOB-7, using polyhydroxyalkanoate (PHA) granules as a solid sustained-release carbon source in RAS was evaluated. With the initial nitrate-N concentration of 140 mg/L, the high denitrification rates of 0.056 g NO3-N L−1 day−1 and 0.035 g NO3-N L−1 day−1 were achieved in denitrification medium containing poly-β-hydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), respectively. Significant erosions and pits formed on the surface of the granules made them a good biofilm carrier for AOB-7, and 3-hydroxybutyrate (3-HB) monomer was the major product released to aquatic phase, which was benefit to animals. SEM photos showed that AOB-7 entered and attached on the inside of the PHA particle holes. A 4-week application trial was conducted to reveal the effects of PHB (AOB-7) denitrifying agent and 3-HB produced on growth of zebrafish (Brachydanio rerio) by adding 0.1% (w/v) PHB (AOB-7) denitrifying agent. Result indicated that PHB (AOB-7) denitrifying agent can significantly reduce nitrate-N content in RASs. Compared with the control group, feed coefficient ratio reduced by 18% and weight gain ratio increased by 29% in the PHB (AOB-7) denitrifying agent group. 3-HB monomer produced during the denitrification was speculated to function as a prebiotic and promote zebrafish growth.

Key Points

AOB-7 showed a good aerobic denitrifying ability on PHA granules as sustained-release C source.

PHB (AOB-7) denitrifying agent can significantly reduce nitrate content in RAS.

R-3-HB monomer was the major product released to aquatic phase and function as a prebiotic.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Behrends A, Klingbeil B, Jendrossek D (1996) Poly (3-hydroxybutyrate) depolymerases bind to their substrate by a C-terminal located substrate binding site. FEMS Microbiol Lett 143:191–194. https://doi.org/10.1111/j.1574-6968.1996.tb08479.x

  2. Correa-Oliveira R, Fachi JL, Vieira A, Sato FT, Vinolo MAR (2016) Regulation of immune cell function by short-chain fatty acids. Clin Transl Immunol 5:e73. https://doi.org/10.1038/cti.2016.17

  3. Delcher AL, Harmon D, Kasif S, White O, Salzberg SL (1999) Improved microbial gene identification with GLIMMER. Nucleic Acids Res 27:4636–4641. https://doi.org/10.1093/nar/27.23.4636

  4. Gao XY, Xu Y, Liu Y, Liu Y, Liu ZP (2012) Bacterial diversity, community structure and function associated with biofilm development in a biological aerated filter in a recirculating marine aquaculture system. Mar Biodivers 42(1):1–11. https://doi.org/10.1007/s12526-011-0086-z

  5. Gutierrez-Wing MT, Malone RF (2006) Biological filters in aquaculture: trends and research directions for freshwater and marine applications. Aquac Eng 34:163–171. https://doi.org/10.1016/j.aquaeng.2005.08.003

  6. Gutierrez-wing MT, Malone RF, Rusch KA (2012) Evaluation of polyhydroxybutyrate as a carbon source for recirculating aquaculture water denitrification. Aquac Eng 51(3):36–43. https://doi.org/10.1016/j.aquaeng.2012.07.002

  7. Hamer HM, Jonkers D, Venema K, Vanhoutvin S, Troost FJ, Brummer RJ (2008) Review article: role of butyrate on colonic function. Aliment Pharmacol Ther 27(2):104–119. https://doi.org/10.1016/j.aquaeng.2012.07.002

  8. Healy MG, Rodgers M, Mulqueen J (2006) Denitrification of a nitrate-rich synthetic wastewater using various wood-based media materials. J Environ Sci Health, Part A: Tox Hazard Subst Environ Eng 41:779–788. https://doi.org/10.1080/10934520600614371

  9. Jendrossek D, Handrick R (2002) Microbial degradation of polyhydroxyalkanoates. Annu Rev Microbiol 56:403–432. https://doi.org/10.1146/annurev.micro.56.012302.160838

  10. Jendrossek D, Schirmer A, Schlegel HG (1996) Biodegradation of polyhydroxyalkanoic acids. Appl Microbiol Biotechnol 46(5–6):451–463. https://doi.org/10.1007/s002530050844

  11. Joo HS, Hirai M, Shoda M (2005) Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis no. 4. J Biosci Bioeng 100:184–191. https://doi.org/10.1263/jbb.100.184

  12. Kaiser H, Schmitz O (1988) Water quality in a closed recirculating fish culture system influenced by addition of a carbon source in relation to feed uptake by fish. Aquac Res 19:265–273. https://doi.org/10.1111/j.1365-2109.1988.tb00429.x

  13. Kim DY, Kim HW, Chung MG, Rhee YH (2007) Biosynthesis, modification, and biodegradation of bacterial medium-chain-length polyhydroxyalkanoates. J Microbiol 45(2):87–97. https://doi.org/10.1016/j.jnt.2007.03.006

  14. Kuwahara A (2014) Contributions of colonic short-chain fatty acid receptors in energy homeostasis. Front Endocrinol (Lausanne) 5:144. https://doi.org/10.3389/fendo.2014.00144

  15. Lee PG, Lea R, Dohmann E, Prebilsky W, Turk P, Ying H, Whitson J (2000) Denitrification in aquaculture systems: an example of a fuzzy logic control problem. Aquac Eng 23:37–59. https://doi.org/10.1016/s0144-8609(00)00046-7

  16. Li L, Gao J, Jiang H, Wang Z (2012) Production of 3-Hydroxybutyrate monomers by Pseudomonas mendocina DS04-T biodegraded polyhydroxybutyrate. J Polym Environ 21(3):826–832. https://doi.org/10.1007/s10924-012-0553-z

  17. Li F, Zhang CY, Liu YT, Liu DB, Xia HM, Chen S (2016) Efficient production of (R)-3-hydroxybutyric acid by Pseudomonas sp. DS1001a and its extracellular poly (3-hydroxybutyrate) depolymerase. Process Biochem 51:369–373. https://doi.org/10.1016/j.procbio.2015.12.016

  18. Lin YF, Jing SR, Wang TW, Lee DY (2002) Effects of macrophytes and external carbon sources on nitrate removal from groundwater in constructed wetlands. Environ Pollut 119:413–420. https://doi.org/10.1016/S0269-7491(01)00299-8

  19. Liu Y, Ai GM, Miao LL, Liu ZP (2016) Marinobacter strain NNA5, a newly isolated and highly efficient aerobic denitrifier with zero N2O emission. Bioresour Technol 206:9–15. https://doi.org/10.1016/j.biortech.2016.01.066

  20. Louis P, Hold GL, Flint HJ (2014) The gut microbiota, bacterial metabolites and colorectal cancer. Nat Rev Microbiol 12:661–672. https://doi.org/10.1038/nrmicro3344

  21. Lowe TM, Eddy SR (1997) tRNAscan-SE: a program for improved detection of transfer RNA genes in genomic sequence. Nucleic Acids Res 25(5):955–964. https://doi.org/10.1093/nar/25.5.0955

  22. Luckachan GE, Pillai CKS (2011) Biodegradable polymers-a review on recent trends and emerging perspectives. J Polym Environ 19(3):637–676. https://doi.org/10.1007/s10924-011-0317-1

  23. Magram SF (2010) Drinking water denitrification in a packed bed anoxic reactor: effect of carbon source and reactor depth. J Appl Sci Res 10:558–563. https://doi.org/10.3923/jas.2010.558.563

  24. Najdegerami EH, Bakhshi F, Tokmechi A, Harzevili AS, Sorgeloos P, Bossier P (2017) Dietary effects of poly-β-hydroxybutyrate on the growth performance, digestive enzyme activity, body composition, mineral uptake and bacterial challenge of rainbow trout fry (Oncorhynchus mykiss). Aquac Nutr 23. https://doi.org/10.1111/anu.12386

  25. Natarajan N, Pluznick JL (2014) From microbe to man: the role of microbial short chain fatty acid metabolites in host cell biology. Am J Physiol Cell Physiol 307(11):C979–C985. https://doi.org/10.1152/ajpcell.00228.2014

  26. Nojiri M, Saito T (1997) Structure and function of poly (3-hydroxybutyrate) depolymerases from Alcaligenes faecalis T1. J Bacteriol 179:6965–6970. https://doi.org/10.1073/pnas.1212591109

  27. Ohura T, Kasuya K, Doi Y (1999) Cloning and characterization of the polyhydroxybutyrate depolymerase gene of Pseudomonas stutzeri and analysis of the function of substrate-binding domains. Appl Environ Microbiol 65:189–197. https://doi.org/10.1109/TAP.2009.2037702

  28. Park W, Jang E, Lee MJ, Yu S, Kim TH (2011) Combination of ion exchange system and biological reactors for simultaneous removal of ammonia and organics. J Environ Manag 92:1148–1153. https://doi.org/10.1016/j.jenvman.2010.11.028

  29. Petersen TN, Brunak S, Heijne GV, Nielsen HH (2011) SIGNALP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8(10):785–786. https://doi.org/10.1038/nmeth.1701

  30. Piazzon MC, Calduch-Giner JA, Fouz B, Estensoro I, Simó-Mirabet P, Puyalto M, Karalazos V, Palenzuela O, Sitjà-Bobadilla A, Pérez-Sánchez J (2017) Under control: how a dietary additive can restore the gut microbiome and proteomic profile, and improve disease resilience in a marine teleostean fish fed vegetable diets. Microbiome 5:164. https://doi.org/10.1186/s40168-017-0390-3

  31. Reddy CSK, Ghai R, Rashmi KVC (2003) Polyhydroxyalkanoates: an overview. Bioresour Technol 87:137–146. https://doi.org/10.1016/s0960-8524(02)00212-2

  32. Schirmer A, Matz C, Jendrossek D (1995) Substrate specificities of PHA-degrading bacteria and active site studies on the extracellular poly(3-hydroxyoctanoic acid) [P(3HO)] depolymerase of Pseudomonas fluorescens GK13. Can J Microbiol 41(Suppl. 1):170–179. https://doi.org/10.1139/m95-184

  33. Shinohe T, Nojiri M, Saito T, Stanislawski T, Jendrossek D (1996) Determination of the active sites serine of the poly(3-hydroxybutyrate) depolymerases of Pseudomonas lemoignei (PhaZ5) and of Alcaligenes faecalis. FEMS Microbiol Lett 141:103–109. https://doi.org/10.1111/j.1574-6968.1996.tb08370.x

  34. Suzuki Y, Taguchi S, Saito S, Toshima K, Matsumura S, Doi Y (2001) Involvement of catalytic amino acid residues in enzyme-catalyzed polymerization for the synthesis of polyesters. Biomacromolecules 2:541–544. https://doi.org/10.1021/bm015508o

  35. Thorburn AN, Macia L, Mackay CR (2014) Diet, metabolites, and “western-lifestyle” inflammatory diseases. Immunity 40(6):833–842. https://doi.org/10.1016/j.immuni.2014.05.014

  36. Van Bussel CGJ, Schroeder JP, Wuertz S, Schulz C (2012) The chronic effect of nitrate on production performance and health status of juvenile turbot (Psetta maxima). Aquaculture 326:163–167. https://doi.org/10.1016/j.aquaculture.2011.11.019

  37. Van Rijn J, Tal Y, Schreier HJ (2006) Denitrification in recirculating systems: theory and applications. Aquac Eng 34:364–376. https://doi.org/10.1016/j.aquaeng.2005.04.004

  38. Wang J, Chu L (2016) Biological nitrate removal from water and wastewater by solid-phase denitrification process. Biotechnol Adv 34:1103–1112. https://doi.org/10.1016/j.biotechadv.2016.07.001

  39. Wang Y, Li F, Wang ZY, Liu DB, Xia HM, Liu LF, Chen S (2012) Purification and properties of an extracellular polyhydroxybutyrate depolymerase from Pseudomonas mendocina DSWY0601. Chem Res Chin Univ 3:459–464. https://doi.org/10.1016/j.mencom.2012.05.021

Download references

Funding

This study was supported by the Biological Resources Program, Chinese Academy of Sciences (KFJ-BRP-009).

Author information

XYG performed the experiments, analyzed the data as well as results, and wrote the manuscript. YL isolated strain AOB-7 used in this work. LLM contributed analytical tools. ZPL supervised all the experiments and revised the manuscript. All authors read and approved the final manuscript.

Correspondence to Zhi-Pei Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. Zebrafish (Brachydanio rerio) was used in the study. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Gao, X., Liu, Y., Miao, L. et al. Pseudomonas sp. AOB-7 utilizes PHA granules as a sustained-release carbon source and biofilm carrier for aerobic denitrification of aquaculture water. Appl Microbiol Biotechnol (2020). https://doi.org/10.1007/s00253-020-10452-y

Download citation

Keywords

  • Denitrification
  • Sustained-release carbon source
  • Biofilm carrier
  • 3-HB monomer