Isolation, identification and application of highly efficient halotolerant strains for coal chemical reverse osmosis concentrate treatment

doi:10.11949/0438-1157.20210212

Abstract

Abstract:

Coal chemical industry plays an important role in national economy of China. Currently, coal chemical wastewater is usually treated by chemical oxidation and biochemical oxidation to remove the organic matter, followed by reverse osmosis (RO) to get the recycled fresh water. The concentrate is subsequently evaporated, while the generated mixture of salts and organic matter belongs to hazardous waste, which is costly to be treated and not environmentally friendly. With the increasing requirements of environmental protection, fractional crystallization has been developed to solve this problem. However, organic matter significantly inhibits salt crystallization. Consequently, there is considerable interest in removing the organic matter of coal chemical RO concentrate efficiently and economically. The cost of advanced oxidation treatment is higher, while it is hard to conduct microbiological treatment in such wastewater since it is weak in high salinity and concentrated toxic compounds. In the work reported here, coal chemical RO concentrate contained 50.9 g/L of TDS and 233.4 mg/L of TOC (total organic carbon) with BOD₅/COD of 0.05. Nine halotolerant bacteria strains were successfully isolated from different sludge and soil, which belonged to Pseudomonas sp., Bacillus sp., and Halomonas sp.by16S rDNA identification. Meanwhile, the research has studied physiological and biochemical characteristics of these nine strains. The results showed that the strains can grow well at 0—15% salinity. Ozone oxidation pretreatment and halotolerant bacteria preparation (mixture of nine strains) treatment was used to treat the wastewater. After one month's continuous operation, the TOC removal ratio reached 40%, reflecting an advanced level compared to the previous study. According to GC-MS (gas chromatography-mass spectrometer) analysis, ozone oxidation can destroy the cyclic structure of organic matter and improve the TOC removal ratio of subsequent halotolerant bacteria preparation. This work provided an effective approach to degrade coal chemical reverse osmosis concentrate by microbial method.

Key words: waste water, degradation, pollution, coal chemical wastewater, reverse osmosis concentrate, halophilic and halotolerant bacteria, halotolerant bacteria preparation, ozone oxidation

摘要：

一般煤化工废水经过多级氧化处理后，反渗透淡水回用、浓水经蒸发产生难处理的“危废”，有机物的存在对“危废”循环利用有显著制约作用。以煤化工反渗透浓水为底物（TOC为233.4 mg/L，TDS为50.9 g/L，BOD₅/COD仅为0.05），从不同菌源中筛选得到9株高效耐盐菌，经16S rDNA测序表明，这些菌株属于假单胞菌属、芽孢杆菌属及嗜盐单胞菌属。将9株耐盐菌配制成复合耐盐菌剂连续式运行处理实际废水，有机物去除率可达30%，为进一步提高去除率，经臭氧氧化预处理，有机物去除率可提高至40%，达到国内外较先进水平。根据气质联用分析，臭氧氧化预处理会破坏废水中环状物质的结构，提高复合耐盐菌剂对难降解有机物的去除效果。本研究为煤化工反渗透浓水中有机物的生物降解提供了可行性方案。

关键词: 废水, 降解, 污染, 煤化工废水, 反渗透浓水, 嗜（耐）盐菌, 复合耐盐菌剂, 臭氧氧化

CLC Number:

X 172

Liting HUANG, Xushen HAN, Yan JIN, Qiang MA, Jianguo YU. Isolation, identification and application of highly efficient halotolerant strains for coal chemical reverse osmosis concentrate treatment[J]. CIESC Journal, 2021, 72(9): 4881-4891.

黄莉婷, 韩昫身, 金艳, 马强, 于建国. 煤化工反渗透浓水的高效降解菌株筛选、鉴定及应用研究[J]. 化工学报, 2021, 72(9): 4881-4891.

Figures/Tables 10

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菌株编号	相似菌株	相似度/%	序列号
L-141	Pseudomonas sp. HC2-4	100.00	JF312947.1
L-142	Pseudomonas sp. strain P0u25	99.92	MK737101.1
L-143	Bacillus sp. (in: Bacteria) strain CY1TSA7	99.79	MH974115.1
L-144	Halomonas alkaliphila X3	100.00	CP024811.1
L-145	Bacillus altitudinis strain HQ-51-Ba	100.00	CP040747.1
L-146	Bacillus altitudinis strain HQ-51-Ba	99.85	CP040747.1
L-147	Bacillus sp. (in: Bacteria) strain CY1TSA7	100.00	MH974115.1
L-275	Bacillus flexus strain HDB-2	99.72	MK178593.1
L-276	Bacillus altitudinis strain NPB34b	100.00	MT598007.1

菌株编号	相似菌株	相似度/%	序列号
L-141	Pseudomonas sp. HC2-4	100.00	JF312947.1
L-142	Pseudomonas sp. strain P0u25	99.92	MK737101.1
L-143	Bacillus sp. (in: Bacteria) strain CY1TSA7	99.79	MH974115.1
L-144	Halomonas alkaliphila X3	100.00	CP024811.1
L-145	Bacillus altitudinis strain HQ-51-Ba	100.00	CP040747.1
L-146	Bacillus altitudinis strain HQ-51-Ba	99.85	CP040747.1
L-147	Bacillus sp. (in: Bacteria) strain CY1TSA7	100.00	MH974115.1
L-275	Bacillus flexus strain HDB-2	99.72	MK178593.1
L-276	Bacillus altitudinis strain NPB34b	100.00	MT598007.1

菌株编号	革兰染色	氧化酶	触酶	淀粉酶	脲酶	吲哚	酪素水解	产H₂S	明胶水解	好氧/厌氧
L-141	-	-	-	+	+	-	+	-	-	+
L-142	+	-	-	-	+	-	+	-	-	+
L-143	+	-	-	-	+	-	+	-	-	+
L-144	+	-	-	-	+	-	+	-	-	+
L-145	+	-	-	-	+	-	+	-	-	+
L-146	+	-	-	-	+	-	+	-	-	+
L-147	+	-	-	-	+	-	+	-	-	+
L-275	+	-	-	-	+	-	+	-	-	+
L-276	+	-	-	+	+	-	+	-	-	+

菌株编号	革兰染色	氧化酶	触酶	淀粉酶	脲酶	吲哚	酪素水解	产H₂S	明胶水解	好氧/厌氧
L-141	-	-	-	+	+	-	+	-	-	+
L-142	+	-	-	-	+	-	+	-	-	+
L-143	+	-	-	-	+	-	+	-	-	+
L-144	+	-	-	-	+	-	+	-	-	+
L-145	+	-	-	-	+	-	+	-	-	+
L-146	+	-	-	-	+	-	+	-	-	+
L-147	+	-	-	-	+	-	+	-	-	+
L-275	+	-	-	-	+	-	+	-	-	+
L-276	+	-	-	+	+	-	+	-	-	+

煤化工反渗透浓水		复合耐盐菌剂法产水		臭氧预处理产水		臭氧预处理和复合耐盐菌剂法联合工艺产水
有机物成分	占比/%	有机物成分	占比/%	有机物成分	占比/%	有机物成分	占比/%
乙二醇单丁醚	2.8	四氧杂环己烷	4.1	乙醇	8.5	5-乙酰氧基-6(1,2-环氧丙基)-5,6二氢吡喃-2-酮	4.5
乙二醇单丁醚	2.8	四氧杂环己烷	4.1	四氧杂环己烷	0.7	5-乙酰氧基-6(1,2-环氧丙基)-5,6二氢吡喃-2-酮	4.5
四氧杂环己烷	1.9	1-亚硝基-2-哌啶甲酸	3.1	二氯异乙醚	40.9	1-甲基-环戊烷羧酸	4.9
1-亚硝基-2-哌啶甲酸	3.9	环己胺	2.5	3-己烯-2,5-二醇	5.5	3-乙基-2-甲基-1-戊烯	5.9
硫代氨基甲酸,N-正环己基S-(2,5-二羟基苯基)	2.7	5-甲基-4-乙烯-3-酮	4.5	2,3-双环呋喃	4.3	2,3-双环呋喃	5.7
2,2-甲基-3-乙烯	6.5	2,3-双环呋喃	4.9	3-乙基-4-甲基-2-戊烯	3.9	2,4-二叔丁基苯酚	9.9
2,3-双环呋喃	9.3	乙基环己烷	1.8	3-乙基-2-戊烯	1.4
5,5-二甲基-3-环己烯-1-醇	23.1	6-exo-vinyl-5-endo-norbornenol	25.9	氯丙醇	0.6
对二氮杂环	2.4	2,6-环辛二烯-1-醇	2.1	二十烷	0.4
2,4-二叔丁基苯酚	2.8			2,4-二叔丁基苯酚	0.7
2,4-二叔丁基苯酚	2.8			二十六烷	0.8