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首页> 外文期刊>Applied Microbiology and Biotechnology >A Pseudomonas putida capable of stereoselective hydrolysis of nitriles
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A Pseudomonas putida capable of stereoselective hydrolysis of nitriles

机译：能够对腈进行立体选择性水解的恶臭假单胞菌

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摘要

Pseudomonas putida NRRL-18668 contains a nitrile hydratase capable of stereoselective hydrolysis of 2-(4-chlorophenyl)-3-methylbutyronitrile at more than 90% enantiomeric excess (ee) to the (S)-amide. This soil isolate was recovered from enrichments using (R,S)-2-methylglutaronitrile as the sole nitrogen source. Enzyme expression is constitutive and does not show a high level of catabolite repression. The organism is capable of growth on a wide variety of aliphatic mono- and dinitrile compounds. The hydrolysis activity on propionitrile is approximately 10.3 mu mole h(-1) mg wet cells(-1). The enzyme in cell-free preparations is inhibited by a number of heavy metals, phenylhydrazine, and cyanide. Substrate specificity is broad with highest rates shown on C-4 and C-5 aliphatic mononitriles. The strain appears somewhat unusual in its dependence on cobalt supplementation for maximum enzyme activity and the ability to hydrolyze some aromatic nitriles. This strain is also capable of a two-step hydrolysis of 2-(4-isobutylphenyl)propionitrile and 2-(6-methoxy-2-napthyl)-propionitrile to the (S)-acids (ibuprofen and naproxen respectively) with stereoselectivity residing primarily in the aliphatic amidase. This appears to be the first description of a steroselective nitrile hydratase from a gram-negative organism.
机译：恶臭假单胞菌NRRL-18668含有腈水合酶，该腈水合酶能够以（S）-酰胺90％以上的对映体过量（ee）立体选择性地水解2-（4-氯苯基）-3-甲基丁腈。使用（R，S）-2-甲基戊二腈作为唯一氮源从浓缩物中回收土壤分离物。酶表达是组成性的，并且不显示高水平的分解代谢物阻遏作用。该生物能够在多种脂族单和二腈化合物上生长。对丙腈的水解活性约为10.3摩尔摩尔h（-1）毫克湿细胞（-1）。无细胞制剂中的酶被多种重金属，苯肼和氰化物抑制。底物特异性广泛，在C-4和C-5脂族单腈上显示出最高的比率。该菌株在最大程度上依赖于钴的补充以获得最大的酶活性和水解某些芳族腈的能力时，显得有些不寻常。该菌株还能够将2-（4-异丁基苯基）丙腈和2-（6-甲氧基-2-萘基）丙腈两步水解为（S）-酸（分别为布洛芬和萘普生），且具有立体选择性。主要存在于脂肪族酰胺酶中。这似乎是对革兰氏阴性生物的立体选择性腈水合酶的首次描述。

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