产多糖根瘤菌Q32对硅酸盐矿物的风化作用及其机制研究
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摘要
微生物与矿物的相互作用是自然界中普遍存在的一种地质作用,可分为直接作用和间接作用两种方式,直接作用是指微生物与矿物直接接触时,在细菌-矿物界面形成有利于矿物风化的微环境,间接作用是指微生物的代谢产物对矿物的化学降解作用。本文研究的主要是微生物与矿物的直接作用,对微生物与矿物相互作用的作用机理进行探讨,尤其是微生物多糖在矿物分解中所起的作用。
供试菌株为实验室保存的一株产胞外多糖根瘤菌Q32,本文在摇瓶培养的条件下,研究了菌株Q32对不同结构硅酸盐矿物的风化作用。结果表明,在0-21d的培养过程中,Q32对钾长石、黑云母、金云母和伊利石均有一定的风化效应。培养21d,接活菌处理发酵液中硅离子含量达到最高,比接灭活菌对照增加138-162%。菌株Q32在以上述硅酸盐矿物为唯一钾源的培养基中生长可形成较多的胞外多糖,且在细菌培养的第7d多糖含量最高,达9.46 g.L-1。
通过扫描电镜(SEM)观察,发现Q32对矿粉颗粒发生了溶蚀作用,矿物表面变得凹凸不平,有腐蚀坑出现,在矿物表面有生物膜形成；能谱(EDS)分析表明,接菌21d后矿物表面的Si、Al含量分别比原始矿物的Si、Al含量低,可以看出矿物上的金属离子被释放出来,Q32对矿物有一定的风化作用。
不同培养条件下Q32所产胞外多糖的分析表明,在培养基加供试矿物的条件下菌株所产多糖与培养基不加矿物所产多糖相比,多糖的含量增加5.8-17.3%,说明矿物能够刺激细菌产糖。气相色谱分析表明,菌株所产多糖为杂多糖,主要是由鼠李糖、木糖、甘露糖、葡萄糖和半乳糖五种单糖组成,从红外图谱上可以找到多糖的特征吸收峰,多糖的红外光谱和单糖组分分析结果表明,在不同条件下所产多糖的结构没有发生明显变化。
纯化后的胞外多糖对不同硅酸盐矿物的溶解试验表明,酸性多糖对矿物有风化作用,主要是因为羧基等酸性基团的螯合作用。多糖中吸附的有机酸有葡萄糖酸,酒石酸和苹果酸,有机酸的浓度分别仅为0.31 mM、0.27 mM和0.29 mM,但低浓度的有机酸并不足以对矿物产生风化作用,本研究表明供试矿物的风化作用主要是由Q32分泌的胞外多糖的溶解作用引起的。
The interaction between minerals and microorganisms is a common geological process in nature. Microorganisms can promote the dissolution of minerals by direct and indirect mechanisms. The direct mechanism refers to the formation of microenvironment in bacteria-mineral interface when microbes and minerals contract directly, which can promote the dissolution of minerals. The indirect mechanism means the chemical degradation effect of metabolites of microbes on minerals. In this study, the role of microbial polysaccharides produced by Rhizobium sp. Q32 in the silicate mineral weathering was investigated.
In this paper, the effect of polysaccharide-producing Rhizobium sp. Q32 on silicate minerals weathering was studied in shaking-flask condition. The result showed that during 21 days of culturing, Rhizobium sp. Q32 could promote the dissolution of K-feldspar, biotite, phlogopite and illite. After 21 days of incubation, the concentration of silicon in solution was increased by 138-162% compared with the control, indicating that the strain Q32 could promote the tested silicate mineral weathering. By using the media containing silicate mineral as the only potassium source, the strain Q32 excreted expolysaccharide, which reached the highest concentration-9.46 mg/L on the 7th day of incubation.
The mineral samples were collected and analzyzed by means of scanning electron microscope (SEM), it was observed that the surfaces of minerals were coated with biofilm and corroded pits were formed on mineral surfaces. Energy dispersive spectronmeter (EDS) images showed that the relative concentrations of Si and Al on the mineral surfaces were lower than those on raw mineral after 21 days of incubation, which demonstrated that metal ions were released from silicate minerals surfaces.
Isolation, purification and analysis of the monosaccharide compositions of expolysaccharide were carried out by the means of UV, IR and GC. The concentration of expolysaccharide produced by strain Q32 in the mineral-containing medium increasd by 5.8-17.3% compared with those in the mineral-free medium, which indicated that minerals can stimulate the the production of expolysacchrides. Rhamnose, Xylose, Galactose, Mannose and Glucose constituted the monosaccharide composition of the expolysacchrides. Characteristic absorption peaks of the polysaccharides could be found from the IR spectra. Analysis of monosaccharide composition of expolysaccharides showed that the structure of the expolysaccharide produced under different contions had not changed.
Purified expolysaccharide could dissolve silicate minerals, mainly due to chelation caused by acid group of the expolysaccharide such as carboxyl. Organic acids such as gluconic acid, tartaric acid and malic acid were aborbed in the expolysaccharide, the concentration of them were only 0.31 mM、0.27 mM and 0.29 mM, respectively. But mineral weathering could not occurred with low concentrations of organic acids, so it indicated that extracellular polysaccharides produced by the strain Q32 played the main role in the mineral weathering in this study.
供试菌株为实验室保存的一株产胞外多糖根瘤菌Q32,本文在摇瓶培养的条件下,研究了菌株Q32对不同结构硅酸盐矿物的风化作用。结果表明,在0-21d的培养过程中,Q32对钾长石、黑云母、金云母和伊利石均有一定的风化效应。培养21d,接活菌处理发酵液中硅离子含量达到最高,比接灭活菌对照增加138-162%。菌株Q32在以上述硅酸盐矿物为唯一钾源的培养基中生长可形成较多的胞外多糖,且在细菌培养的第7d多糖含量最高,达9.46 g.L-1。
通过扫描电镜(SEM)观察,发现Q32对矿粉颗粒发生了溶蚀作用,矿物表面变得凹凸不平,有腐蚀坑出现,在矿物表面有生物膜形成；能谱(EDS)分析表明,接菌21d后矿物表面的Si、Al含量分别比原始矿物的Si、Al含量低,可以看出矿物上的金属离子被释放出来,Q32对矿物有一定的风化作用。
不同培养条件下Q32所产胞外多糖的分析表明,在培养基加供试矿物的条件下菌株所产多糖与培养基不加矿物所产多糖相比,多糖的含量增加5.8-17.3%,说明矿物能够刺激细菌产糖。气相色谱分析表明,菌株所产多糖为杂多糖,主要是由鼠李糖、木糖、甘露糖、葡萄糖和半乳糖五种单糖组成,从红外图谱上可以找到多糖的特征吸收峰,多糖的红外光谱和单糖组分分析结果表明,在不同条件下所产多糖的结构没有发生明显变化。
纯化后的胞外多糖对不同硅酸盐矿物的溶解试验表明,酸性多糖对矿物有风化作用,主要是因为羧基等酸性基团的螯合作用。多糖中吸附的有机酸有葡萄糖酸,酒石酸和苹果酸,有机酸的浓度分别仅为0.31 mM、0.27 mM和0.29 mM,但低浓度的有机酸并不足以对矿物产生风化作用,本研究表明供试矿物的风化作用主要是由Q32分泌的胞外多糖的溶解作用引起的。
The interaction between minerals and microorganisms is a common geological process in nature. Microorganisms can promote the dissolution of minerals by direct and indirect mechanisms. The direct mechanism refers to the formation of microenvironment in bacteria-mineral interface when microbes and minerals contract directly, which can promote the dissolution of minerals. The indirect mechanism means the chemical degradation effect of metabolites of microbes on minerals. In this study, the role of microbial polysaccharides produced by Rhizobium sp. Q32 in the silicate mineral weathering was investigated.
In this paper, the effect of polysaccharide-producing Rhizobium sp. Q32 on silicate minerals weathering was studied in shaking-flask condition. The result showed that during 21 days of culturing, Rhizobium sp. Q32 could promote the dissolution of K-feldspar, biotite, phlogopite and illite. After 21 days of incubation, the concentration of silicon in solution was increased by 138-162% compared with the control, indicating that the strain Q32 could promote the tested silicate mineral weathering. By using the media containing silicate mineral as the only potassium source, the strain Q32 excreted expolysaccharide, which reached the highest concentration-9.46 mg/L on the 7th day of incubation.
The mineral samples were collected and analzyzed by means of scanning electron microscope (SEM), it was observed that the surfaces of minerals were coated with biofilm and corroded pits were formed on mineral surfaces. Energy dispersive spectronmeter (EDS) images showed that the relative concentrations of Si and Al on the mineral surfaces were lower than those on raw mineral after 21 days of incubation, which demonstrated that metal ions were released from silicate minerals surfaces.
Isolation, purification and analysis of the monosaccharide compositions of expolysaccharide were carried out by the means of UV, IR and GC. The concentration of expolysaccharide produced by strain Q32 in the mineral-containing medium increasd by 5.8-17.3% compared with those in the mineral-free medium, which indicated that minerals can stimulate the the production of expolysacchrides. Rhamnose, Xylose, Galactose, Mannose and Glucose constituted the monosaccharide composition of the expolysacchrides. Characteristic absorption peaks of the polysaccharides could be found from the IR spectra. Analysis of monosaccharide composition of expolysaccharides showed that the structure of the expolysaccharide produced under different contions had not changed.
Purified expolysaccharide could dissolve silicate minerals, mainly due to chelation caused by acid group of the expolysaccharide such as carboxyl. Organic acids such as gluconic acid, tartaric acid and malic acid were aborbed in the expolysaccharide, the concentration of them were only 0.31 mM、0.27 mM and 0.29 mM, respectively. But mineral weathering could not occurred with low concentrations of organic acids, so it indicated that extracellular polysaccharides produced by the strain Q32 played the main role in the mineral weathering in this study.
引文
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