Screening of an Efficient Denitrifying Phosphorus-accumulating Bacterium and Its Denitrification and Phosphorus Removal

doi:10.13560/j.cnki.biotech.bull.1985.2022-1522

Abstract

Abstract:

The objective is to explore the denitrification and phosphorus removal conditions and performance of a efficient denitrifying phosphorus-accumulating bacterium, and its treatment effect on simulated wastewater. Denitrifying phosphorus-accumulating strain D4 was isolated by pure culture technology. Preliminary identification of the strain was completed based on 16S rRNA gene sequence analysis and physiological and biochemical characteristics, and the denitrification and dephosphorization conditions were optimized by single factor and response surface method, so as to achieve efficient biological denitrification and dephosphorization of mimic food wastewater. As results，denitrifying phosphorus-accumulating bacterium D4 was Gordonia sp.. The optimal conditions for nitrogen and phosphorus removal were sodium acetate（3.32 g/L）as carbon source, 5% inoculum, initial phosphorus content of 18.61 mg/L, 31.3℃, and pH 7.9. Under these conditions, the removed amounts of total phosphorus, nitrate nitrogen and ammonia nitrogen by strain D4 were 14.18, 39.67 and 69.71 mg/L, respectively; and the corresponding removal rates were 84.50%, 97.67% and 96.22%. Results of treatment to different simulated wastewater showed that strain D4 had the best nitrogen and phosphorus removal effects in soybean product wastewater, with the removed amount of nitrogen and phosphorus being 101.58 mg/L and 10.40 mg/L, respectively. In conclusion, strain D4 has promising denitrification and phosphorus-accumulating ability and is a good strain resource for biological denitrification and phosphorus removal of food processing wastewater with high nitrogen and phosphorus such as soybean product wastewater.

Key words: denitrification and phosphorus-accumulating bacterium, denitrification and phosphorus removal, condition optimization, wastewater treatment

YUAN Ye, ZHOU Jia, QU Jian-hang, ZHANG Bo-yuan, LUO Yu, LI Hai-feng. Screening of an Efficient Denitrifying Phosphorus-accumulating Bacterium and Its Denitrification and Phosphorus Removal[J]. Biotechnology Bulletin, 2023, 39(7): 266-276.

Figures/Tables 12

Table 1 Three factors and three levels of denitrifying phosphorus-accumulating bacteria

因素代码 Code of factor	因素 Factor	水平Level
因素代码 Code of factor	因素 Factor	1	0	1
A	温度 Temperature/℃	20	28	34
B	pH	7	8	9
C	初始磷含量Initial phosphorus content/（mg·L^-1）	10	15	20

Fig. 1 Phylogenetic tree based on the 16S rRNA genes of strain D4 Code in parentheses is GenBank accession number of strain.The number of branch nodes is bootstrap value（That of lower than 70 is not displayed）. The data of the scale is the evolutionary distance

Fig. 2 Growth and nitrogen and phosphorus removal curves of strain D4

Fig. 3 Effects of carbon sources and carbon source concentration on the nitrogen and phosphorus removal efficiency of strain D4 Different lower letters indicate significant difference at P<0.05 level. The same below

Fig. 4 Effects of inoculum size on nitrogen and phosphorus removal efficiency of D4

Fig. 5 Effects of initial phosphorus concentration on the nitrogen and phosphorus removal efficiency of D4

Fig. 6 Effects of temperature on the nitrogen and phosphorus removal efficiency of D4

Fig. 7 Effects of pH on the nitrogen and phosphorus removal efficiency of D4

Table 2 Significance analysis results

因素Source	平方和Sum of squares	自由度Freedom	均方Mean square	F值F value	显著性Significance
碳源Carbon source	143.696	4	35.924	376.090	0.000**
乙酸钠浓度Sodium acetate concentration	43.381	4	10.845	583.658	0.000**
接种量Vaccination percent	0.064	4	0.016	0.535	0.714
初始磷含量Initial phosphorus content	49.980	4	12.495	114.939	0.000**
温度Temperature	75.747	4	18.937	288.322	0.000**
pH	150.384	4	37.593	1492.194	0.000**

Table 3 Analysis of variance of quadratic model

来源Source	平方和Sum of squares	自由度Freedom	均方Mean square	F值F value	P值P value	显著性Significance
模型	139.61	9	15.51	22.05	0.0002	**
A	15.89	1	15.89	22.59	0.0021	**
B	2.02	1	2.02	2.88	0.1337	-
C	53.54	1	53.54	76.12	< 0.0001	**
AB	2.52	1	2.52	3.58	0.1002	-
AC	9.49	1	9.49	13.50	0.0079	**
BC	0.0038	1	0.0038	0.0055	0.9431	-
A²	13.90	1	13.90	19.75	0.0030	**
B²	8.09	1	8.09	11.51	0.0116	*
C²	8.81	1	8.81	12.53	0.0095	**
残值	4.92	7	0.7034
失拟项	3.68	3	1.23	3.96	0.1083	-
纯误差	1.24	4	0.3099
总计	144.54	16

Fig. 8 Response surface diagram of interaction of various factors on total phosphorus removal a: The interaction of temperature and pH; b: the interaction of temperature and initial phosphorus concentration; c: the interaction of pH and initial phosphorus concentration

Fig. 9 Removed amount of total phosphorus (a), total nitrogen (b), ammonia nitrogen (c) in different simulated wastewater by bacterial strain D4

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