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Widespread and active piezotolerant microorganisms mediate phenolic compound degradation under high hydrostatic pressure in hadal trenches
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作者 Hao Ling yongxin lv +2 位作者 Yu Zhang Ning‑Yi Zhou Ying Xu 《Marine Life Science & Technology》 SCIE CSCD 2024年第2期331-348,共18页
Phenolic compounds,as well as other aromatic compounds,have been reported to be abundant in hadal trenches.Although high-throughput sequencing studies have hinted at the potential of hadal microbes to degrade these co... Phenolic compounds,as well as other aromatic compounds,have been reported to be abundant in hadal trenches.Although high-throughput sequencing studies have hinted at the potential of hadal microbes to degrade these compounds,direct microbiological,genetic and biochemical evidence under in situ pressures remain absent.Here,a microbial consortium and a pure culture of Pseudomonas,newly isolated from Mariana Trench sediments,efficiently degraded phenol under pressures up to 70 and 60 MPa,respectively,with concomitant increase in biomass.By analyzing a high-pressure(70 MPa)culture metatranscriptome,not only was the entire range of metabolic processes under high pressure generated,but also genes encod-ing complete phenol degradation via ortho-and meta-cleavage pathways were revealed.The isolate of Pseudomonas also contained genes encoding the complete degradation pathway.Six transcribed genes(dmpKLMNOP_(sed))were functionally identified to encode a multicomponent hydroxylase catalyzing the hydroxylation of phenol and its methylated derivatives by heterogeneous expression.In addition,key catabolic genes identified in the metatranscriptome of the high-pressure cultures and genomes of bacterial isolates were found to be all widely distributed in 22 published hadal microbial metagenomes.At microbiological,genetic,bioinformatics,and biochemical levels,this study found that microorganisms widely found in hadal trenches were able to effectively drive phenolic compound degradation under high hydrostatic pressures.This information will bridge a knowledge gap concerning the microbial aromatics degradation within hadal trenches. 展开更多
关键词 Hadal trench High hydrostatic pressure Phenolic compounds degradation Piezotolerant microorganisms Widespread distribution
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基于内源Ⅲ-B型CRISPR-Cas系统构建超嗜热火球菌Pyrococcus yayanosii A1的基因敲降系统
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作者 陈柔珂 吕永新 +2 位作者 张欢欢 宋庆浩 徐俊 《微生物学报》 CAS CSCD 北大核心 2021年第7期1920-1932,共13页
【目的】亚氏火球菌(Pyrococcus yayanosii)A1菌株的最适生长温度为98℃,最适生长压力为5.2×10^(4) Pa,是研究此类严格厌氧、超嗜热和兼性嗜压古菌的环境适应性机制的良好材料。本研究基于P.yayanosii A1基因组中Ⅲ-B型CRISPR-Cas... 【目的】亚氏火球菌(Pyrococcus yayanosii)A1菌株的最适生长温度为98℃,最适生长压力为5.2×10^(4) Pa,是研究此类严格厌氧、超嗜热和兼性嗜压古菌的环境适应性机制的良好材料。本研究基于P.yayanosii A1基因组中Ⅲ-B型CRISPR-Cas系统,旨在建立可应用于此类古菌的基因表达敲降系统(gene knock-down system)。【方法】人工构建的mini-CRISPR簇,由内源性CRISPR簇Group 1的重复序列(repeats)和待敲降的淀粉酶基因(PYCH_13690)中的原间隔序列(protospacer)组成。将该mini-CRISPR簇插入到具有辛伐他汀抗性的穿梭载体pLMOShhp中,使之转录与目的基因mRNA匹配的crRNA,引导Cmr复合物对其进行切割。【结果】使用高静水压(HHP)诱导型启动子Phhp诱导mini-CRISPR簇表达时,淀粉酶基因的mRNA数量在5.2×10^(4) Pa静水压时下调到原来的67.05%,在1.0×10^(2) Pa静水压时下调为原来的49.69%;而使用组成型启动子Phmtb诱导mini-CRISPR簇表达时,淀粉酶基因的mRNA数量在5.2×10^(4) Pa静水压时下调为原来的58.48%,在1.0×10^(2) Pa静水压时下调为原来的23.97%。使用鲁戈氏碘液显色法对这两类基因表达敲降突变菌株的培养物中残留的淀粉含量进行分析,结果显示突变菌株的淀粉降解能力明显降低。【结论】在P.yayanosii A1中建立了基于内源Ⅲ-B型CRISPR-Cas系统的基因敲降系统,可以用于抑制此类超嗜热嗜压古菌体内基因的表达。 展开更多
关键词 超嗜热古菌 嗜压微生物 PYROCOCCUS yayanosii CRISPR-Cas 基因敲降
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