摘要
利用变色圈法从孔雀石绿污染的养殖池塘中筛选到1株具有较强孔雀石绿脱色和降解能力的细菌,经过16S rDNA基因序列比对及系统进化树分析,结合VITEK 2微生物鉴定系统,鉴定该菌为Klebsiella pneumoniae,重命名为K.pneumoniae WA-1。采用分光光度法和硼氢化钾还原-高效液相色谱法研究不同碳源对孔雀石绿脱色和降解的影响,发现菌株在盐培养基中表现出很强的孔雀石绿脱色和降解能力,添加葡萄糖、半乳糖、蔗糖等碳源时孔雀石绿(5~40 mg/L)的脱色率有所提高,均接近100%,但是孔雀石绿(5 mg/L)降解率显著降低。明确了孔雀石绿首先在细胞外由孔雀石绿还原酶还原成隐性孔雀石绿,然后通过主动运输进入到细胞被进一步降解的脱色和降解过程,并揭示了不同碳源对菌株孔雀石绿脱色和降解的影响机制。本研究为K.pneumoniae WA-1的孔雀石绿降解机制研究提供重要思路,也为后期孔雀石绿降解的应用提供重要理论基础。
In this study,a strain with strong ability to decolorize and degrade malachite green was isolated from a malachite green-contaminated aquaculture pond by the color-changing zone method.By 16S rDNA sequence alignment and phylogenetic tree analysis,as well as using the microbial identification system VITEK 2,the strain was identified as Klebsiella pneumoniae and named as WA-1.The effects of different carbon sources on decolorization and degradation of malachite green by WA-1 were studied by spectrophotometry and KBH4 reduction-high performance liquid chromatography with fluorescence detection(HPLC-FLD).The results showed that this strain potently decolorized and degraded malachite green in the salt medium.The decolorization rate of malachite green at 5–40 mg/L was increased to approximately 100%after the addition of carbon sources including glucose,galactose,and sucrose,while the degradation rate was significantly reduced,especially at 5 mg/L.Malachite green was reduced to leucomalachite green by malachite green reductase outside the cells,which could enter the cells through active transport for decolorization and degradation.The mechanism of the effect of different carbon sources on the decolorization and degradation of malachite green was also revealed.The results of this study provide an important idea to reveal the mechanism of malachite green degradation by K.pneumoniae WA-1,and also provide an important theoretical basis for the application of malachite green degradation in the future.
作者
田小丽
李春生
陈胜军
薛勇
邓建朝
潘创
王悦齐
TIAN Xiaoli;LI Chunsheng;CHEN Shengjun;XUE Yong;DENG Jianchao;PAN Chuang;WANG Yueqi(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,China;Key Laboratory of Aquatic Product Processing,Ministry of Agriculture and Rural Affairs,National R&D Center for Aquatic Product Processing,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2022年第6期181-188,共8页
Food Science
基金
现代农业(特色淡水鱼)产业技术体系建设专项(CARS-46)
中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项(2020XK01)
广东省现代农业产业技术体系创新团队建设专项(2022KJ151)
广东省重点领域研发计划项目(2019B020225001)
中国水产科学研究院基本科研业务费专项(2020TD69)。
