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首页> 外文期刊>Biotechnology Progress >Stress survival of a genetically engineered Pseudomonas in soil slurries:Cytochrome P-450cam-catalyzed dehalogenation of chlorinated hydrocarbons
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Stress survival of a genetically engineered Pseudomonas in soil slurries:Cytochrome P-450cam-catalyzed dehalogenation of chlorinated hydrocarbons

机译：基因工程假单胞菌在土壤泥浆中的应力存活：细胞色素P-450cam催化氯代烃的脱卤

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摘要

Biological treatment of hazardous chemical wastes has potential as an effective, practical, and economically viable process in above the ground treatment systems that consist of both genetically engineered microorganisms (GEMs) and bioreactors with process control instruments to create ideal conditions for biodegradation. A strain of Pseudomonas putida coexpressing cytochrome P-450cam and luciferase (lux) that provides both the reductive detoxification potential of the hemoprotein and a mechanism for its reduction in the absence of "normal" P-450 redox partners was evaluated for its ability to survive and remain metabolically competent under nutrient stress in soil slurry microcosms. More than 74% of the cells of engineered Pseudomonas were culturable after 7 days of multiple nutrient (C,N,P) starvation. The diagnostic luminescence and carbon monoxide-difference spectra for the two engineered traits could be detected in a significant fraction of the surviving population. The GEM could be revived after repeated desiccation and starvation using Luria broth, benzoate, or citrate as nutrients. Soil slurries inoculated with the GEM transformed hexachloroethane (HCE) to tetrachloroethylene (tetraCE) 8-10-fold faster than uninoculated slurries. The GEM also transformed the insecticide, gamma-HCH (gamma-3,4,5,6-hexachlorocyclhexene), to gamma-3,4,5,6-tetrachlorocyclohexene (gamma tetraCH) in soil slurries under subatmospheric conditions. These results indicate that GEMs can be;constructed with broad substrate range detoxification catalysts such as cytochrome P-450 for remediation.
机译：在地面处理系统（包括基因工程微生物（GEM）和生物反应器）以及地面处理系统组成的地面处理系统中，危险化学废物的生物处理具有潜力，可以作为一种有效，实用且经济上可行的方法，从而为生物降解创造理想的条件。评价了共表达细胞色素P-450cam和荧光素酶（lux）的恶臭假单胞菌菌株，该菌株既可提供血红蛋白的还原性解毒潜能，又可提供在不存在“正常” P-450氧化还原伴侣的情况下可降低其毒理的机制并在养分胁迫下保持土壤泥浆微观世界的代谢能力。经过多次营养（C，N，P）饥饿7天后，可以培养超过74％的工程化假单胞菌细胞。这两个工程性状的诊断发光和一氧化碳差异光谱可在幸存的人口中检测到。使用Luria肉汤，苯甲酸盐或柠檬酸盐作为营养物反复干燥和饥饿后，GEM可以恢复。用GEM接种的土壤泥浆比未接种的泥浆快8-10倍，将六氯乙烷（HCE）转化为四氯乙烯（tetraCE）。创业板还在低于大气压的条件下，将杀虫剂γ-六氯环己烷（γ-3,4,5,6-六氯环己烯）转化为γ-3,4,5,6-四氯环己烯（γtetraCH）。这些结果表明GEMs可以用宽范围的底物范围的解毒催化剂如细胞色素P-450进行修复。

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