产微生物絮凝剂的菌株筛选、鉴定及发酵条件优化
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摘要
微生物絮凝剂是一类新型的水处理制剂,因其具有高效、无毒、易降解等特点,已成为水处理剂开发的研究热点。为了扩大微生物絮凝剂的种类,本研究从活性污泥和土壤样品中筛选絮凝剂产生菌,最终获得一株具有絮凝活性的菌株,通过16S rDNA鉴定,该菌株为蒙氏假单胞菌(Pseudomonas monteilii),因此命名为P. monteilii YR-1。利用单因素实验和正交试验对该菌株的发酵条件进行了优化,其最佳碳、氮源分别为25 g/L葡萄糖和2 g/L复合氮源(酵母浸粉:尿素:硫酸铵=5:5:2),培养基初始pH值为10,培养温度为28℃,发酵时间为72 h。在最佳培养条件下,其发酵液的絮凝率从37.68%提高到63.52%。这是关于蒙氏假单胞菌产絮凝剂的首次报道。
Microbial flocculant(MBF) is a new type of water treatment agent, which has become a research hotspot of water treatment agent development with characteristics of high efficiency, nontoxicity and easy degradation. In order to expand the types of MBF, a strain with flocculation activity was finally obtained from activated sludge and soil samples. The strain was identified as Pseudomonas monteilii by 16 S r DNA, and named P. monteilii YR-1. The fermentation conditions of the strain were optimized by single factor experiment and orthogonal test. The optimum carbon and nitrogen sources were 25 g/L glucose and 2 g/L complex nitrogen sources(yeast extract fermentation: urea:ammonium sulfate=5: 5: 2). The initial p H of the culture medium was 10, the culture temperature was 28℃ and the fermentation time was 72 h. Under the optimum conditions, the flocculation rate of fermentation broth increased from37.68% to 63.52%. This was the first report on the production of flocculants by P. monteilii.
Microbial flocculant(MBF) is a new type of water treatment agent, which has become a research hotspot of water treatment agent development with characteristics of high efficiency, nontoxicity and easy degradation. In order to expand the types of MBF, a strain with flocculation activity was finally obtained from activated sludge and soil samples. The strain was identified as Pseudomonas monteilii by 16 S r DNA, and named P. monteilii YR-1. The fermentation conditions of the strain were optimized by single factor experiment and orthogonal test. The optimum carbon and nitrogen sources were 25 g/L glucose and 2 g/L complex nitrogen sources(yeast extract fermentation: urea:ammonium sulfate=5: 5: 2). The initial p H of the culture medium was 10, the culture temperature was 28℃ and the fermentation time was 72 h. Under the optimum conditions, the flocculation rate of fermentation broth increased from37.68% to 63.52%. This was the first report on the production of flocculants by P. monteilii.
引文
Aljuboori A.H.R.,Idris A.,Abdullah N.,and Mohamad R.,2013,Production and characterization of a bioflocculant produced by Aspergillus flavus,Bioresource Technology,127:489-493
Buthelezi S.P.,Olaniran A.O.,and Pillay B.,2012,Textile dye removal from wastewater effluents using bioflocculants produced by indigenous bacterial isolates,Molecules,17(12):14260-14274
Fu L.,Jiang B.,Liu J.,Zhao X.,Liu Q.,and Hu X.,2016,Genome sequence analysis of a flocculant-producing bacterium,Paenibacillus shenyangensis,Biotechnology Letters,38(3):447-453
Gomaa E.Z.,2012,Production and characteristics of a heavy metals removing bioflocculant produced by Pseudomonas aeruginosa,Polish Journal of Microbiology,61(4):281-289
Li O.,Lu C.,Liu A.,Zhu L.,Wang P.M.,Qian C.D.,Jiang X.H.,and Wu X.C.,2013,Optimization and characterization of polysaccharide-based bioflocculant produced by Paenibacillus elgii B69 and its application in wastewater treatment,Bioresource Technology,134:87-93
Shi C.F.,and Leng X.Y.,2015,Treatment of pharmaceutical wastewater by microbial flocculant,Xiandai Huagong(Modern Chemical Industry),35(9):85-89(石春芳,冷小云,2015,微生物絮凝剂在制药废水处理中的应用研究,现代化工,35(9):85-89)
Salehizadeh H.,and Shojaosadati S.A.,2001,Extracellular biopolymeric flocculants:recent trends and biotechnological importance,Biotechnology Advances,19(5):371-385
Salehizadeh H.,and Yan N.,2014,Recent advances in extracellular biopolymer flocculants,Biotechnology Advances,32(8):1506-1522
Xu C.H.,and Yu L.,2017,Screening of producing strain of microbial flocculant and optimization of culture conditions,Yinran Zhuji(Textile Auxiliaries),34(7):11-15(徐长绘,于兰,2017,微生物絮凝剂产生菌的筛选及培养条件的优化,印染助剂,34(7):11-15)
Yang J.,Wu D.,Li A.,Guo H.,Chen H.,Pi S.S.,Wei W.,and Ma F.,2016,The addition of N-hexanoyl-homoserine lactone to improve the microbial flocculant production of Agrobacterium tumefaciens Strain F2,an exopolysaccharide bioflocculantproducing bacterium,Applied Biochemistry and Biotechnology,179(5):728-739
Yuan W.T.,Jiao Y.C.,Wang J.,and Wang H.,2017,Identification of flocculant-producing microbia l JMH 48 and optimization of its flocculant producing conditions,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),36(1):252-258(元王涛,焦迎春,王婧,王虹,2017,絮凝菌JMH 48的鉴定及其产絮凝剂条件的优化研究,基因组学与应用生物学,36(1):252-258)
Yang Q.,Luo K.,Liao D.X.,Li X.M.,Wang D.B.,Liu X.,Zeng G.M.,and Li X.,2012,A novel bioflocculant produced by Klebsiella sp.and its application to sludge dewatering,Water and Environment Journal,26(4):560-566
Zhang J.W.,2017,The production of microbial flocculants,Guangdong Huagong(Guangdong Chemical Industry),44(8):71-75(张静雯,2017,微生物絮凝剂的研制,广东化工,44(8):71-75)
Zhao C.,Yang Q.,and Zhang H.,2017,Optimization of microbial flocculant-producing medium for Bacillus subtilis,Indian Journal Microbiology,57(1):83-91
Zhao H.J.,Liu H.T.,and Zhou J.G.,2013,Characterization of a bioflocculant MBF-5 by Klebsiella pneumoniae and its application in Acanthamoeba cysts removal,Bioresource Technology,137:226-232
Zhuang X.L.,Wang Y.P.,Li Q.B.,Yan S.,and He N.,2012,The production of bioflocculants by Bacillus licheniformis using molasses and its application in the sugarcane industry,Biotechnology Bioprocess Engineering,17(5):1041-1047
Buthelezi S.P.,Olaniran A.O.,and Pillay B.,2012,Textile dye removal from wastewater effluents using bioflocculants produced by indigenous bacterial isolates,Molecules,17(12):14260-14274
Fu L.,Jiang B.,Liu J.,Zhao X.,Liu Q.,and Hu X.,2016,Genome sequence analysis of a flocculant-producing bacterium,Paenibacillus shenyangensis,Biotechnology Letters,38(3):447-453
Gomaa E.Z.,2012,Production and characteristics of a heavy metals removing bioflocculant produced by Pseudomonas aeruginosa,Polish Journal of Microbiology,61(4):281-289
Li O.,Lu C.,Liu A.,Zhu L.,Wang P.M.,Qian C.D.,Jiang X.H.,and Wu X.C.,2013,Optimization and characterization of polysaccharide-based bioflocculant produced by Paenibacillus elgii B69 and its application in wastewater treatment,Bioresource Technology,134:87-93
Shi C.F.,and Leng X.Y.,2015,Treatment of pharmaceutical wastewater by microbial flocculant,Xiandai Huagong(Modern Chemical Industry),35(9):85-89(石春芳,冷小云,2015,微生物絮凝剂在制药废水处理中的应用研究,现代化工,35(9):85-89)
Salehizadeh H.,and Shojaosadati S.A.,2001,Extracellular biopolymeric flocculants:recent trends and biotechnological importance,Biotechnology Advances,19(5):371-385
Salehizadeh H.,and Yan N.,2014,Recent advances in extracellular biopolymer flocculants,Biotechnology Advances,32(8):1506-1522
Xu C.H.,and Yu L.,2017,Screening of producing strain of microbial flocculant and optimization of culture conditions,Yinran Zhuji(Textile Auxiliaries),34(7):11-15(徐长绘,于兰,2017,微生物絮凝剂产生菌的筛选及培养条件的优化,印染助剂,34(7):11-15)
Yang J.,Wu D.,Li A.,Guo H.,Chen H.,Pi S.S.,Wei W.,and Ma F.,2016,The addition of N-hexanoyl-homoserine lactone to improve the microbial flocculant production of Agrobacterium tumefaciens Strain F2,an exopolysaccharide bioflocculantproducing bacterium,Applied Biochemistry and Biotechnology,179(5):728-739
Yuan W.T.,Jiao Y.C.,Wang J.,and Wang H.,2017,Identification of flocculant-producing microbia l JMH 48 and optimization of its flocculant producing conditions,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),36(1):252-258(元王涛,焦迎春,王婧,王虹,2017,絮凝菌JMH 48的鉴定及其产絮凝剂条件的优化研究,基因组学与应用生物学,36(1):252-258)
Yang Q.,Luo K.,Liao D.X.,Li X.M.,Wang D.B.,Liu X.,Zeng G.M.,and Li X.,2012,A novel bioflocculant produced by Klebsiella sp.and its application to sludge dewatering,Water and Environment Journal,26(4):560-566
Zhang J.W.,2017,The production of microbial flocculants,Guangdong Huagong(Guangdong Chemical Industry),44(8):71-75(张静雯,2017,微生物絮凝剂的研制,广东化工,44(8):71-75)
Zhao C.,Yang Q.,and Zhang H.,2017,Optimization of microbial flocculant-producing medium for Bacillus subtilis,Indian Journal Microbiology,57(1):83-91
Zhao H.J.,Liu H.T.,and Zhou J.G.,2013,Characterization of a bioflocculant MBF-5 by Klebsiella pneumoniae and its application in Acanthamoeba cysts removal,Bioresource Technology,137:226-232
Zhuang X.L.,Wang Y.P.,Li Q.B.,Yan S.,and He N.,2012,The production of bioflocculants by Bacillus licheniformis using molasses and its application in the sugarcane industry,Biotechnology Bioprocess Engineering,17(5):1041-1047