Characterization of the Propham Biodegradation Pathway in Starkeya sp. Strain YW6 and Cloning of a Novel Amidase Gene mmH
Characterization of the Propham Biodegradation Pathway in Starkeya sp. Strain YW6 and Cloning of a Novel Amidase Gene mmH
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Starkeya sp 中 Propham 生物降解途径的表征。
DOI：
10.1021/acs.jafc.8b06928
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发表时间：
2019
影响因子：
6.1
通讯作者：
Liu Hongming
中科院分区：
文献类型：
--
作者：
Sun Lina;Gao Xinhua;Chen Wei;Huang Kaihua;Bai Naling;Lyu Weiguang;Liu Hongming
我们之前分离出了一株降解单色磷的菌株starkeyasp。YW6，也可以降解苯胺。在这里，我们发现菌株YW6通过一种途径代谢苯胺，苯胺首先被水解成苯胺，然后转化为儿茶酚，儿茶酚随后通过异性裂解途径被氧化。从菌株YW6中克隆出新的氨基酶基因，并在大肠杆菌bl21 （DE3）中表达。具有芳基酰基酰胺酶活性的MmH被纯化用于酶分析。生物信息学分析证实，MmH属于酰胺酶特征（AS）酶家族，与多个AS家族成员具有26-50%的同源性。MmH（分子量为53 kDa）在40°C和pH 8.0条件下活性最强，对苯胺的kcat1和kkm值分别为33.4 s - 1和16.9 μM。这些特性使得MmH适合用于新型酰胺的生物合成和环境修复。
We previously isolated a monocrotophos-degrading strainStarkeyasp. YW6, which could also degrade propham. Here, we show that strain YW6 metabolizes propham via a pathway in which propham is initially hydrolyzed to aniline and then converted to catechol, which is then oxidized via anortho-cleavage pathway. The novel amidase genemmHwas cloned from strain YW6 and expressed inEscherichia coliBL21(DE3). MmH, which exhibits aryl acylamidase activity, was purified for enzymatic analysis. Bioinformatic analysis confirmed that MmH belongs to the amidase signature (AS) enzyme family and shares 26–50% identity with several AS family members. MmH (molecular mass of 53 kDa) was most active at 40 °C and pH 8.0 and showed high activity toward propham, withKcatandKmvalues of 33.4 s–1and 16.9 μM, respectively. These characteristics make MmH suitable for novel amide biosynthesis and environmental remediation.