脱氮硫杆菌胞内胞外酶对硫酸盐还原菌酶活抑制机理研究

Sulfate reducing bacteria (SRB) is one of the most main kinds of anaerobic bacteria recognized lead to corrosion of metal material by the researchers at home and abroad. It is very important that using the characteristics of microbe inhibit the growth of SRB in the metal material corrosion and environmental protection. The Ideas and plans that intracellular and extracellular enzymes of thiobacillus denitrification(TDN) inhibit intracellular enzymes activities of SRB are proposed. A research method that SRB enzymes activities are determined is adopted in this project. The influence of TDN intracellular and extracellular enzymes to SRB growth and its enzymes activities is explored. In this project, the main research contents are as followed. (1) TDN is cultivated and TDN intracellular and extracellular enzymes are separated and purified. (2) SRB is cultivated and SRB intracellular enzymes are separated and purified. (3) Inhibition behaviors of TDN enzymes to SRB bacteria are determined. (4) Inhibition behaviors of TDN enzymes to SRB enzymes are determined. (5)Whether SRB enzymes are inhibited by TDN enzymes will be demonstrated. Therefore, in enzymes activity inhibition mechanism, not only nitrogen oxides from TDN, but also TDN enzymes are explored to inhibit SRB enzymes activities. Perfect enzyme activity inhibition mechanism will provide theoretical guidance to the selection and application of anticorrosion microbe and have scientific significance in the field of microbial corrosion and protection research.

硫酸盐还原菌（SRB）是目前国内外学者公认的导致金属材料腐蚀最主要的一类细菌。利用微生物的特性抑制SRB的生长在金属材料的防腐及保护环境方面非常重要。本项目提出了对脱氮硫杆菌（TDN）胞内酶和胞外酶能否抑制SRB胞内酶活性进行论证的思路与方案。将采用检测SRB酶活的研究方法，探究TDN酶对SRB生长及其酶活的影响，实现对酶活抑制机理的补充和完善。本项目重点研究内容包括：①培养TDN，从中分离纯化得到TDN胞内酶和胞外酶；②培养SRB，从中分离纯化得到SRB胞内酶；③测试TDN胞内酶和胞外酶对微生物SRB的抑制效果；④测试TDN胞内酶和胞外酶对SRB胞内酶酶活的抑制效果；⑤论证TDN酶能否抑制SRB酶活，使酶活抑制机理不仅考虑TDN代谢物氮氧化物，而且也考虑TDN酶对SRB酶代谢过程的抑制作用。完善的酶活抑制机理将对防腐微生物的遴选与应用提供理论指导，在微生物防腐研究领域具有科学意义。

硫酸盐还原菌（Sulfate-reducing bacteria，SRB）是目前学者公认的导致金属材料腐蚀最主要的一类细菌。利用微生物的特性抑制SRB的生长在金属材料的防腐及保护环境方面非常重要。脱氮硫杆菌（T. denitrificans, TD）是硫杆菌属中的一类兼性厌氧菌，短杆状，以极生鞭毛运动，广泛存在于土壤、污泥和淡水等环境中。我们实验室研究发现，TDN能够有效抑制SRB对EH36钢的腐蚀。本项目对SRB和TDN进行发酵培养，通过高速离心、超声破壁、丙酮沉淀等方法得到胞内胞外酶，测定结果表明TDN胞内酶能抑制SRB胞内酶和胞外酶的酶活性。通过电化学阻抗谱法、生物检测技术和X射线光电子能谱法等，对TDN胞内酶抑制SRB腐蚀EH36钢的行为进行研究。结果表明，SRB胞内粗酶液对EH36钢产生腐蚀比较严重，纯化后的TDN胞内酶能抑制SRB胞内酶对EH36钢腐蚀。本研究有助于完善酶活性抑制机制，且能为防腐微生物的选择和应用提供理论指导，在微生物防腐研究领域具有重要的科学意义。