Metaproteomic insights into ammonia oxidising bacterial consortium developed for bioaugmenting nitrification in aquacult
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ORIGINAL ARTICLE
Metaproteomic insights into ammonia oxidising bacterial consortium developed for bioaugmenting nitrification in aquaculture systems Divya Jose 1 & Prasannan Geetha Preena 1 & Vattiringal Jayadradhan Rejish Kumar 1,2 & Rosamma Philip 3 & Isaac Sarojini Bright Singh 1 Received: 5 July 2019 / Accepted: 26 March 2020 # Institute of Molecular Biology, Slovak Academy of Sciences 2020
Abstract The ammonia oxidizing bacterial consortium (AMOPCU-1) has been under use for efficient biological removal of ammonia from the recirculating aquaculture system. In this context a metaproteomic approach to the study of the consortium was undertaken to analyse the proteins involved in nitrification. Ammonia removal efficiency was monitored while amplifying the consortium in 2 L fermenter under standard conditions, and the consortium kept under substrate starvation served as the control. When the bacterial consortium attained peak ammonia removal efficiency, their total proteins were extracted. Protein profiling was done using one dimensional and two-dimensional SDS-PAGE. Eleven visible protein spots, which appeared during peak ammonia oxidation were identified by MALDI-TOF analysis. The proteins could be linked to metabolic processes that were active during nitrification. These included enzymes that influenced ammonia oxidation by regulating intracellular cytoplasmic pH of bacterial cells and transcription of ammonia monooxygenase, proteins having key roles in electron transport chain, ATP generation and fatty acid synthesis, and bacterial signal proteins that respond to environmental nitrite and nitrate and nitrous oxide reductase, a key enzyme in complete nitrate reduction. The present study is the first of its kind undertaken to investigate the metaproteome of ammonia oxidising consortium through which relevant proteins involved in efficient nitrification-denitrification could be revealed. Keywords Ammonia oxidising . Nitrification . Nitrosomonas . Nitrobacter . Metaproteomics
Abbreviations RAS AMO NOR FISH qPCR SDS-PAGE
MALDI-TOF-MS Recirculating Aquaculture Systems Ammonia monooxygenase Nitrite Oxidoreductase Fluorescence in situ Hybridisation Quantitative PCR Sodium Dodecyl Sulfate–Poly Acrylamide Gel Electrophoresis
* Isaac Sarojini Bright Singh [email protected] 1
National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Lakeside Campus, Fine Arts Avenue, Cochin 682016, India
2
Present address: Department of Aquaculture, Kerala University of Fisheries and Ocean Studies, Panangad, Kochi 682506, India
3
Department of Marine biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology, Lakeside Campus, Fine Arts Avenue, Cochin 682016, India
2-D-PAGE IEF NCBI
Matrix-assisted laser desorption/ionization – Time of Flight-Mass Spectrometry Two- dimensional-Poly Acrylamide Gel Electrophoresis Isoelectric Focussing National Center for Biotechnology Information
Introduction Accumulation of ammonia and nitrite to toxic levels is
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