Hlighted for several biotechnological Imiquimod 99011-02-6 applications. An attempt is biodegradation of toxic chemicals such as herbicides, xenobiotics in the environment. Since acyl anilide herbicides in the soil environment, bacteria AAA, by attacking the amide bonds in chemicals, reducing their concentrations in the soil and k can be used Will pollute for bioremediation. Another application is the detection of S. acetaminophenol in clinical trials. pa acetaminophenol an amide bond cozy Vinegar Acid and p aminophenol. Since the AAA acts only on non-peptide amide bonds, the bioassay was proposed p acetamido phenol in biological fluids using AAA and as a template for a biosensor system. The other application is the synthesis of an aryl acylamide with the reverse reaction of the AAA, which a method for preparing biocatalytic substance acylamide invention Can Participate en aryl. A familiar example is the synthesis of aniline analgesics of AAA. Here pr We will present a new AAA bacterial gene in a genomic library from a soil bacterium that is isolated, identified a media selection acetaminophenol with p as the sole carbon source, and deposited in GenBank. Sequence analysis revealed that the gene in the family as an enzyme rt go. We studied the evolution Re relationship with other known enzymes and classified into six families through her mind, such as regions. To examine the biochemical properties, we overexpressed the Irbesartan 138402-11-6 gene in E. coli and examined the properties and the kinetic parameters of the enzyme gene on various substrates, indicating a m Aligned for biotechnological applications using the gene AAA. Materials and Methods bacterial strains Strains and plasmids We used EE coli NM522 and DH5 coli for the construction of a genomic library and as h Their general cloning. E. coli BL21 was used as the h For protein expression. To construct the genomic library, we used pBluescript II KS. A modified cloning vector pET21a independent Independent ligation for the production of recombinant proteins, which was kindly provided by Berkeley Center for Structural Genomics are provided. EE coli NM522 and were routinely coli DH5 Strength at 37 in Luria Bertani cultured medium with ampicillin, and E. coli BL21, the recombinant plasmid was grown to 25, 5052 ZYM automatic inducing medium with ampicillin, as claimed in claim Studier protocol BSGC provided. Cloning a gene of arylacylamidase genomic library from a soil bacterium was isolated a soil bacterium, as described in a previous study. Briefly, the methods as follows. Soil samples were collected from different areas in South Korea. The samples were dissolved in saline Suspended solution and filtered through Whatman No. 1 filter paper. The filtrates were in selection medium, inoculated containing 0.1% w acetaminophenol as the sole carbon source. A single isolate was separated from the plate Doxorubicin selection medium, acid bacteria after a series of transfers of lactic. To a genomic library for screening of a gene of aryl acyl Amidaseaktivit To construct t, we extracted chromosomal DNA using a commercial kit. Genomic DNA was digested with BamHI and treated with pBluescript II KS with the same restriction enzyme with T4 ligase to according to the manufacturer’s protocol. The library was screened in LB medium.