Isolation, Identification, Denitrification Characteristics of a Highly Efficient Aerobic Denitrifying Bacterial Strain DS2
WANG He1,2, YI Meng-Meng2, WANG Miao2, GAO Feng-Ying2, KE Xiao-Li2, CAO Jian-Meng2, LIU Zhi-Gang2, LU Mai-Xin2,*
1 College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China; 2 Pearl River Fisheries Research Institute, Chinese Academy of Fishery Science/Key Laboratory of Tropical and Subtropical Fishery Resource Application and Cultivation, Ministry of Agriculture/Guangdong Provincial Key Laboratory of Aquatic Animal Immune Technology, Guangzhou 510380, China
Abstract：Excess nitrogen in the aquaculture waterbodies has caused deterioration of the aquaculture environment, and seriously affects the benefits of aquaculture. Microbial denitrification is one of the most economical and effective measures for the treatment of nitrogenous pollutants. In this study, a high-efficiency aerobic denitrifying bacteria DS2 (GenBank No. PRJNA838789) was isolated from the biofilter filler of circulating water aquaculture system. Strain DS2 was identified as Pseudomonas putida based on physiological and biochemical characteristics, and analysis of 16S rRNA gene sequence. Single-factor experiment, denitrification experiment, and whole-genome sequencing technology were used to analyze the optimal growth conditions, denitrification performance and mechanism of strain DS2. The results showed that the strain grew fastest under the condition of 30 ℃, pH7, 250 r/min, and salinity of 10‰~20‰. After 24 h of cultivation in heterotrophic nitrification medium (HNM), short-cut denitrification medium (SDM), denitrification medium (DM), and simultaneous nitrification and denitrification medium (SNDM), the total nitrogen (TN) removal efficiency of strain DS2 was 93.50%, 55.60%, 65.98%, 90.10%, respectively. Moreover, the functional genes related to nitrogen metabolism were defined in the genome of strain DS2, such as narB, nirK, nirA, norB, and nirB. Finally, in aquaculture wastewater treatment experiment, strain DS2 obtained 99.72% removal rate of nitrate and 89.52% removal rate of TN in 12 h. The present study indicates that Pseudomonas putida strain DS2 is a high efficiency aerobic denitrifying bacterium, and possesses great potential for application in aquaculture wastewater treatment.
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