絮凝剂产生菌GA1及所产絮凝剂MBFGA1的研究
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摘要
微生物絮凝剂是微生物在生长过程中所产生的代谢产物,其主要成分为多糖、糖蛋白、蛋白质、纤维素、核酸等。与其它絮凝剂相比,微生物絮凝剂具有种类多、来源广、培养条件粗放、絮凝性能好、无环境毒性等特点。由于传统的絮凝剂存在不同的缺陷,尤其是铝系絮凝剂和聚丙烯酰胺类絮凝剂具有环境毒性。因此,微生物絮凝剂自从问世以来便受到广泛的关注,具有重要的研究价值和应用前景。
多粘类芽孢杆菌Paenibacillus polymyxa GA1为本实验室从土壤中筛选出来的一株高效产絮菌株,所产絮凝剂MBFGA1具有广泛的絮凝、脱色作用。本论文针对GA1培养特性以及所产絮凝剂MBFGA1产生情况、絮凝性能等方面展开了广泛深入的研究,主要工作有:(1)考察了MBFGA1理化性质、絮凝效果、影响因素;(2)考察了GA1发酵培养基以及各组分的对絮凝的影响,并深入研究影响机理;(3)研究磷酸盐对GA1培养过程参数、MBFGA1化学构成和产生情况的影响及机制;(4)研究了GA1菌株和MBFGA1对重金属铜离子的去除情况。
研究表明,MBFGA1主要成分为多糖,热稳定性良好;GA1发酵液在CaCl_2存在条件下和偏碱时对高岭土、土壤废水、洗煤废水等絮凝效果达到95%以上,但对垃圾渗滤液处理效果相对较差,絮凝率为58%左右;
培养基中磷酸盐在CaCl_2存在下,pH值中性偏碱时对各种废水均表现一定的絮凝性,在MBFGA1絮凝废水的过程中起重要作用,且作用离子为PO_4~(3-)和Ca2+;这两种离子在pH值中性偏碱时能有效降低高龄土悬液的ξ电位绝对值到接近零,使胶体脱稳;PO_4~(3-)和Ca~(2+)+形成絮状沉淀后,能网捕脱稳颗粒,进而在MBFGA1桥联作用下颗粒结团沉降完成絮凝;发酵培养基虽有一定絮凝性,但效果不佳,处理后上清液颗粒平均粒径远大于发酵液处理后,且絮体、泥饼性质差异大,不利于工业应用和后续处理;BaSO4亦有一定的助凝效果,但因沉淀本身性质差异,其效果劣于磷酸钙。
磷酸盐对GA1的生长曲线阶段划分影响较小,但能有效促进对数生长速率和最终生物量;磷酸盐非MBFGA1合成的必要条件,但能促进产絮快速启动,缩短产品收获时间,提高最终单位产率,其他条件相同时,含有磷酸盐的培养基最终单位产率比不含磷酸盐的培养基高30%;磷酸盐对体系pH值变化具有缓冲作用,能维持pH值相对稳定,有利于GA1菌体的生长;含有磷酸盐的培养基和不含磷酸盐的培养基产生的MBFGA1均为多糖结构,区别在于前者结构中不含叁键和累积双键,而后者含有R-C≡CH。
GA1菌体对重金属铜离子有一定的吸附去除效果,且吸附平衡时间较短,为8min。吸附作用为表面吸附,不参与能量和物质代谢;不同生长阶段的GA1对铜离子去除效果差异较大,静止期内,吸附效果微弱,之后吸附率快速上升,在对数尾期达最高值74%,高于用发酵液处理时的最高值;MBFGA1对铜离子有一定的去除作用,但效果较弱,最大负载为1 mg/ 3mg;pH值对发酵液吸附效果影响较大,pH为6时吸附效果最佳,为74%。
Bioflocculants, having good flocculating activity, are the metabolic products of the microorganism, which are safe, non-toxic , easily cultivated, and can be biodegraded. Its main components are amylose, glycoprotein, nucleic acid, protein etc. Since the advantages of using bioflocculants in both environmental safety and economic over some other inorganic flocculants and organic flocculants, great attention has been payed to explore the bioflocculants mechanism and to investigate their characteristics in use. As a result, it is of great value in the research of bioflocculant.
Paenibacillus polymyxa GA1 was a bacteria with high bioflocculant producing rate which was screened from soil by our lab. It could produce bioflocculant MBFGA1 with superordinary flocculability and decolourising property.This study focused on flocculability of MBFGA1and the situation of GA1’s growth. The works remained: (1) research into the characteristic ,flocculability and influencing factors of MBFGA1; (2) study on the influence and mechanism of culture medium and its ingredients to flocculability; (3) study on the influence of phosphate to the parameter in culturing, chemical structure and producing situation; (4) study on the adsorption of Cu2+.
The study showed, MBFGA1’s main component was polysaccharide, and its thermal stability was superordinary. With CaCl_2, when pH>7,the flocculating rate of kaolin, soil waste water and coal waste water were above 95%,but only about 58% of landfill leachate.
Different wastewater samples could be flocculated by phosphate in the culture medium with CaCl2 when pH >7.0. K_2HPO_4 and KH_2PO_4 were the key factors when the wastewater was treated by the microbial flocculant produced by GA1(MBFGA1).The contributing ions were PO_4~(3-) and Ca~(2+). The absolute value of kaolin’s zeta potential could be brought down by these two ions to about 0, hence the stabilization of the colloid was broken. The unstable grains could be captured by the floccule of calcium phosphate, then the grains coagulated with MBFGA1 and settled. Although the wastewater could be flocculated by the culture medium alone, the effect was poor. The average granularity of kaolin was much bigger than that treated by fermentation and the floc properties are inappropriate for industrial conditions. Kaolin could also be flocculated with the help of BaSO4, but because of the difference of deposition, the effect was not as good as Ca3(PO4)2.
The growth velocity in logarithm-time and the final productivity could be increased because of Phosphate,but there was no difference in division of cell growth curve with or without Phosphate. Phosphate was not necessary when MBFGA1 was produced,but the start-point of producing could be earlier with it. The final unit productivity with phosphate was 30% higher then without phosphate when the other factors were the same; The phosphate was some kind of buffering agent which made the pH steady.Steady pH was good for the cell growth. Both the MBFGA1with phosphate or without phosphate were polysaccharide, the only difference in structure was that there was R-C≡CH in the former but not in the latter.
Paenibacillus polymyxa GA1 could adsorbed Cu~(2+), and the equilibration time was 8 min. This adsorption effect was surface adsorption, Cu~(2+) did not participate the energy and principle metabolism. The adsorption effect of GA1 in different phase was different, the effect of GA1 in stationary phase was poor, and than the effect was rising faster,the best adsorption effect occurred in the phase of last logarithm-time,the adsorption rate was 74%.
多粘类芽孢杆菌Paenibacillus polymyxa GA1为本实验室从土壤中筛选出来的一株高效产絮菌株,所产絮凝剂MBFGA1具有广泛的絮凝、脱色作用。本论文针对GA1培养特性以及所产絮凝剂MBFGA1产生情况、絮凝性能等方面展开了广泛深入的研究,主要工作有:(1)考察了MBFGA1理化性质、絮凝效果、影响因素;(2)考察了GA1发酵培养基以及各组分的对絮凝的影响,并深入研究影响机理;(3)研究磷酸盐对GA1培养过程参数、MBFGA1化学构成和产生情况的影响及机制;(4)研究了GA1菌株和MBFGA1对重金属铜离子的去除情况。
研究表明,MBFGA1主要成分为多糖,热稳定性良好;GA1发酵液在CaCl_2存在条件下和偏碱时对高岭土、土壤废水、洗煤废水等絮凝效果达到95%以上,但对垃圾渗滤液处理效果相对较差,絮凝率为58%左右;
培养基中磷酸盐在CaCl_2存在下,pH值中性偏碱时对各种废水均表现一定的絮凝性,在MBFGA1絮凝废水的过程中起重要作用,且作用离子为PO_4~(3-)和Ca2+;这两种离子在pH值中性偏碱时能有效降低高龄土悬液的ξ电位绝对值到接近零,使胶体脱稳;PO_4~(3-)和Ca~(2+)+形成絮状沉淀后,能网捕脱稳颗粒,进而在MBFGA1桥联作用下颗粒结团沉降完成絮凝;发酵培养基虽有一定絮凝性,但效果不佳,处理后上清液颗粒平均粒径远大于发酵液处理后,且絮体、泥饼性质差异大,不利于工业应用和后续处理;BaSO4亦有一定的助凝效果,但因沉淀本身性质差异,其效果劣于磷酸钙。
磷酸盐对GA1的生长曲线阶段划分影响较小,但能有效促进对数生长速率和最终生物量;磷酸盐非MBFGA1合成的必要条件,但能促进产絮快速启动,缩短产品收获时间,提高最终单位产率,其他条件相同时,含有磷酸盐的培养基最终单位产率比不含磷酸盐的培养基高30%;磷酸盐对体系pH值变化具有缓冲作用,能维持pH值相对稳定,有利于GA1菌体的生长;含有磷酸盐的培养基和不含磷酸盐的培养基产生的MBFGA1均为多糖结构,区别在于前者结构中不含叁键和累积双键,而后者含有R-C≡CH。
GA1菌体对重金属铜离子有一定的吸附去除效果,且吸附平衡时间较短,为8min。吸附作用为表面吸附,不参与能量和物质代谢;不同生长阶段的GA1对铜离子去除效果差异较大,静止期内,吸附效果微弱,之后吸附率快速上升,在对数尾期达最高值74%,高于用发酵液处理时的最高值;MBFGA1对铜离子有一定的去除作用,但效果较弱,最大负载为1 mg/ 3mg;pH值对发酵液吸附效果影响较大,pH为6时吸附效果最佳,为74%。
Bioflocculants, having good flocculating activity, are the metabolic products of the microorganism, which are safe, non-toxic , easily cultivated, and can be biodegraded. Its main components are amylose, glycoprotein, nucleic acid, protein etc. Since the advantages of using bioflocculants in both environmental safety and economic over some other inorganic flocculants and organic flocculants, great attention has been payed to explore the bioflocculants mechanism and to investigate their characteristics in use. As a result, it is of great value in the research of bioflocculant.
Paenibacillus polymyxa GA1 was a bacteria with high bioflocculant producing rate which was screened from soil by our lab. It could produce bioflocculant MBFGA1 with superordinary flocculability and decolourising property.This study focused on flocculability of MBFGA1and the situation of GA1’s growth. The works remained: (1) research into the characteristic ,flocculability and influencing factors of MBFGA1; (2) study on the influence and mechanism of culture medium and its ingredients to flocculability; (3) study on the influence of phosphate to the parameter in culturing, chemical structure and producing situation; (4) study on the adsorption of Cu2+.
The study showed, MBFGA1’s main component was polysaccharide, and its thermal stability was superordinary. With CaCl_2, when pH>7,the flocculating rate of kaolin, soil waste water and coal waste water were above 95%,but only about 58% of landfill leachate.
Different wastewater samples could be flocculated by phosphate in the culture medium with CaCl2 when pH >7.0. K_2HPO_4 and KH_2PO_4 were the key factors when the wastewater was treated by the microbial flocculant produced by GA1(MBFGA1).The contributing ions were PO_4~(3-) and Ca~(2+). The absolute value of kaolin’s zeta potential could be brought down by these two ions to about 0, hence the stabilization of the colloid was broken. The unstable grains could be captured by the floccule of calcium phosphate, then the grains coagulated with MBFGA1 and settled. Although the wastewater could be flocculated by the culture medium alone, the effect was poor. The average granularity of kaolin was much bigger than that treated by fermentation and the floc properties are inappropriate for industrial conditions. Kaolin could also be flocculated with the help of BaSO4, but because of the difference of deposition, the effect was not as good as Ca3(PO4)2.
The growth velocity in logarithm-time and the final productivity could be increased because of Phosphate,but there was no difference in division of cell growth curve with or without Phosphate. Phosphate was not necessary when MBFGA1 was produced,but the start-point of producing could be earlier with it. The final unit productivity with phosphate was 30% higher then without phosphate when the other factors were the same; The phosphate was some kind of buffering agent which made the pH steady.Steady pH was good for the cell growth. Both the MBFGA1with phosphate or without phosphate were polysaccharide, the only difference in structure was that there was R-C≡CH in the former but not in the latter.
Paenibacillus polymyxa GA1 could adsorbed Cu~(2+), and the equilibration time was 8 min. This adsorption effect was surface adsorption, Cu~(2+) did not participate the energy and principle metabolism. The adsorption effect of GA1 in different phase was different, the effect of GA1 in stationary phase was poor, and than the effect was rising faster,the best adsorption effect occurred in the phase of last logarithm-time,the adsorption rate was 74%.
引文
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