Novel phthalate hydrolase EstJ6 as well as coding gene and application thereof

A phthalate ester and hydrolase technology, applied in the fields of hydrolase, application, genetic engineering, etc., can solve the problems of low efficiency of new esterases, non-cultivation, and restrictions on the development and utilization of new enzymes
CN110982803AActive Publication Date: 2020-04-10NANJING AGRICULTURAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NANJING AGRICULTURAL UNIVERSITY
Publication Date
2020-04-10

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Abstract

The invention provides a novel phthalate hydrolase gene derived from a soil metagenome library. The nucleotide sequence and an amino acid sequence of the novel phthalate hydrolase gene are shown as SEQ ID NO.1 and SEQ ID NO.2. After the esterase gene is connected to an expression vector pET28a (+), the esterase gene is transformed into escherichia coli BL21 (DE3) to realize heterologous expression. The molecular weight of the purified recombinase (EstJ6) is 33.31 kDa. The EstJ6 has wide substrate specificity on phthalate, and the EstJ6 not only can hydrolyze phthalate with a simple side chain,but also can hydrolyze diethyl hexyl phthalate and monoethyl hexyl phthalate with complex and longer side chains. In addition, site-specific mutagenesis experiments show that the catalytic triad residue of EstJ6 is S146-E240-H270, and mutation of any amino acid in the three causes the EstJ6 to lose the catalytic ability. The novel phthalate hydrolase disclosed by the invention can be applied to the fields of food industry, agriculture, biotechnology and the like due to the specific activity and enzymatic characteristics of the novel phthalate hydrolase.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a novel phthalate hydrolase obtained by screening from a soil metagenomic library, its coding gene and its application. Background technique

[0002] Phthalates (PAEs), a class of toxic organic compounds, are ubiquitous in the environment due to their widespread use as additives or plasticizers in the manufacture of plastics. Six of them (dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), dioctyl phthalate (DNOP), phthalate Diethylhexyl dicarboxylate (DEHP) and butylbenzyl phthalate (BBP) have been listed as priority pollutants by the US Environmental Protection Agency and some of their international counterparts. Among them, DEHP occupies a high proportion (about 31%) in the environment. The massive use of PAEs has resulted in serious environmental problems and a variety of diseases, including respiratory diseases, childhood obesity, and neurop...

Claims

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