一株链球菌产微生物絮凝剂的研究
摘要
微生物絮凝剂是天然高分子絮凝剂的重要种类。它具有无毒、无二次污染的显著特点，而且在絮凝沉降的同时还具有可生化降解等功能。产絮凝剂的微生物种类多，生长快，易于采取生物方法实现产业化，具有广阔的发展前途。
从福州郊区地表土壤中分离得到一株絮凝剂产生菌，其菌落具有表面光滑、呈透明的蛋清状、形状较大的特点。筛选结果表明：从产荚膜微生物的菌落形态来初步筛选、分离絮凝剂产生菌的方法是可行性的，并具有快速和简便的优点。同时也证明荚膜与微生物絮凝活性存在一定的联系。微生物鉴定结果为链球菌属。
在实验室条件下确立了该菌株的发酵培养基组成和最适摇瓶发酵条件及絮凝剂使用的实验室最佳条件。对高岭土的絮凝实验研究表明：金属阳离子能影响絮凝剂的絮凝活性。结合离子形态、特性和有机高分子絮凝的化学模型，初步分析了该种微生物絮凝剂可能的絮凝机理。
对发酵获得的絮凝活性物质进行了分离和提取。应用DEAE-Sephadex A25，Sephdarose 4B凝胶色谱柱从发酵液中提取活性物质并纯化。通过TLC、GC、UV、IR、元素分析等表征后初步推测了其主要成分为酸性多糖，其中多糖含量(73％)、糖醛酸含量(20.6％)、多糖分子量为1.0×10~(5.63)Da，可能还有少量氨基或短肽结构。
应用该絮凝剂处理南安市某石板材厂废水，结果表明：微生物絮凝剂能有效去除石材废水中的固体颗粒悬浮物。在废水处理过程中少量Ca~(2+)和搅拌能有效降低出水的浊度，去除率可达83％。
Bioflocculant is an important sort of the macromolecular flocculants. It has such advantages: no toxicity, no pollution. Bioflocculant produced by microorganism can be easy to achieve industrialization through biology engineering and biodecomposed.
The microorganism that can produce bioflocculant was screened by conventional method from the soil. One streptococcus strain possess capsule was separated completely. The result indicates: the method observing the community shapes was simple and convenient for screening flocculants producing microorganisms.
The studies were focused on the optimum culture medium, culture conditions and apply conditions. The investigation of flocculating the alkaline kaolin suspensions suggests that metal ions can affect the flocculate rate distinctly. A likely model was put forward based on experiment phenomena.
Ethanol and Sevag precipitated the bioflocculant that was purified to homogeneity through Sephadex-DEAE A-25 and Sepharose 4B gel chromatogram columns. The results suggested the effective component in bioflocculant was polysaccharide characterized by TLC GC UV IR and elemental analysis. The biopolymer contained sugar (73%) and uronic acid (20.6%), elemental analysis suggested little amdio and peptide might exist. The molecular weight measured by HPLC was 1.0 X 105.63Da.
The flocculation effect, SS was removed distinctly; of bioflocculant on the treatment of lapis waste water was observed. Turbidity analysis indicated additional Ca2+ and agitation would improve the water quality. Turbidity Removal efficiency is up to 83%.
从福州郊区地表土壤中分离得到一株絮凝剂产生菌，其菌落具有表面光滑、呈透明的蛋清状、形状较大的特点。筛选结果表明：从产荚膜微生物的菌落形态来初步筛选、分离絮凝剂产生菌的方法是可行性的，并具有快速和简便的优点。同时也证明荚膜与微生物絮凝活性存在一定的联系。微生物鉴定结果为链球菌属。
在实验室条件下确立了该菌株的发酵培养基组成和最适摇瓶发酵条件及絮凝剂使用的实验室最佳条件。对高岭土的絮凝实验研究表明：金属阳离子能影响絮凝剂的絮凝活性。结合离子形态、特性和有机高分子絮凝的化学模型，初步分析了该种微生物絮凝剂可能的絮凝机理。
对发酵获得的絮凝活性物质进行了分离和提取。应用DEAE-Sephadex A25，Sephdarose 4B凝胶色谱柱从发酵液中提取活性物质并纯化。通过TLC、GC、UV、IR、元素分析等表征后初步推测了其主要成分为酸性多糖，其中多糖含量(73％)、糖醛酸含量(20.6％)、多糖分子量为1.0×10~(5.63)Da，可能还有少量氨基或短肽结构。
应用该絮凝剂处理南安市某石板材厂废水，结果表明：微生物絮凝剂能有效去除石材废水中的固体颗粒悬浮物。在废水处理过程中少量Ca~(2+)和搅拌能有效降低出水的浊度，去除率可达83％。
Bioflocculant is an important sort of the macromolecular flocculants. It has such advantages: no toxicity, no pollution. Bioflocculant produced by microorganism can be easy to achieve industrialization through biology engineering and biodecomposed.
The microorganism that can produce bioflocculant was screened by conventional method from the soil. One streptococcus strain possess capsule was separated completely. The result indicates: the method observing the community shapes was simple and convenient for screening flocculants producing microorganisms.
The studies were focused on the optimum culture medium, culture conditions and apply conditions. The investigation of flocculating the alkaline kaolin suspensions suggests that metal ions can affect the flocculate rate distinctly. A likely model was put forward based on experiment phenomena.
Ethanol and Sevag precipitated the bioflocculant that was purified to homogeneity through Sephadex-DEAE A-25 and Sepharose 4B gel chromatogram columns. The results suggested the effective component in bioflocculant was polysaccharide characterized by TLC GC UV IR and elemental analysis. The biopolymer contained sugar (73%) and uronic acid (20.6%), elemental analysis suggested little amdio and peptide might exist. The molecular weight measured by HPLC was 1.0 X 105.63Da.
The flocculation effect, SS was removed distinctly; of bioflocculant on the treatment of lapis waste water was observed. Turbidity analysis indicated additional Ca2+ and agitation would improve the water quality. Turbidity Removal efficiency is up to 83%.
引文
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[2] 马青山，等．絮凝化学与絮凝剂．北京．中国环境科学出版社．1988
[3] 茹志刚．废水治理工程技术．北京．中国环境科学出版社．1989
[4] 陈坚．环境生物技术应用与发展．北京．中国轻工业出版社．2001．6
[5] 仓根隆一郎．微生物的研究状况及应用前景．日本化学会志．1992，(5)：453-463
[6] 李为群，刘健，赵宇华．高分子絮凝剂开发应用新动向．环境污染与防治．1997，19(3)：32-36
[7] 刘杏，邵林广，张丽芳．生物絮凝剂的研究现状及发展趋势．孝感学院学报．2001，21(6)：39-42
[8] 陈元彩，肖锦．天然有机高分子絮凝剂研究与应用．环境科学进展．199917(3)：84-89
[9] S.P. Vijayalakshmi, A.M. Raichur. the utility of Bacillus a bioflocculant for fine coal. Colloids and Surfaces B:Biointerfaces. 2003, 29: 265-275
[10] 吴健，戴桂馥．微生物细胞的絮凝与微生物絮凝剂．环境污染与防治．1994，16(6)：27-30
[11] KuraneR. Screening for and Characteristics of Microbial Flocculants. Biol. Chem. 1986,50(9): 2301-2307
[12] Jin Sik Nam. Bioflocculant produced by Aspergillus SE JS-42. Biosci Biotech Bioehem. 1996,60(2): 325-327
[13] Kurane R. Screening and characteristics of microbial flocculants. Agric Biol Chem. 1986,50(9): 2301-2307
[14] Zolotuchin I. Biological flocculant of suspended substance in mine waters. Chimija Technologija. 1990,12(9): 855-857
[15] Takagi.H, Kadowaki.K. Flocculant produced by Paecilomyces SP taxono
mic studies and culture conditions for production. Agric BioChem. 1985, 49(5): 55-58
[16] 仓根隆一郎．Screening,isolation and some properties of microbial cell flocculants. PPM. 1992, (10): 44-51
[17] 胡勇有，高健：微生物絮凝剂的研究与应用进展．环境污染治理技术与设备．1999，7(4)：24-29
[18] 王镇，王孔星，谢裕敏，姚茵丽．几株絮凝剂产生菌的特性研究．微生物学报．1995，35(2)：121-129
[19] 庄益源，戴树桂，宋文华．新型絮凝剂凝集水中染料的研究．环境工程与生态工程．1997，1：1-3
[20] Salehizadeh.H. Extracellular biopolymeric flocculants Recent trends and biotechnological importance. Biotechnology Advances. 2001,19: 317-385
[21] 叶晶菁，谭天伟．微生物絮凝剂产生菌分离选育及提取鉴定．北京化工大学学报．2001，28(1)：11-17
[22] 肖琳．高效微生物絮凝剂产生菌X-1的分离和絮凝性研究．第一届全国环境化学学术讨论会论文集
[23] Kurane R, Matsuyama H. Production of a bioflocculant by mixed culture. Biotechnol, Biochem. 1994,(58): 253-257
[24] ShihI.L. Production of biopolymer flocculant from bacillus licheniformis and its flocculantion properties. Bioresource Technology. 2001,78:267-272
[25] 张通，卢文玉，田春．絮凝剂产生菌培养基的研究．应用与环境生物学报．2003，9(1)：67-70
[26] 何宁，李寅，陆茂林．营养和环境条件对生物絮凝剂合成的影响．应用与环境生物学报．2001，7(5)：483-488
[27] Jang Jin-Ho. Production of a novel bioflocculant by fed-batch culture of Citrobacter sp. Biotechnology Letters. 2001, 23:593-597
[28] 王镇．几株絮凝剂产生菌的特性研究．微生物学报．1995，35：122-129
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