Dehalogenase HldD1, encoding gene thereof and application of dehalogenase HldD1

A dehalogenase and encoding technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of toxicity, environmental and health consequences, and seriousness of microorganisms

Active Publication Date: 2021-04-20
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI +1
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Problems solved by technology

However, because the stability of halogenated organic substances is generally high, and they are even toxic to microorganisms that may degrade them, the biodegradability of halogenated organic substances has been poor for many years.
Excessive exposure to these halogenated pollutants can lead to serious environmental and health consequences

Method used

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  • Dehalogenase HldD1, encoding gene thereof and application of dehalogenase HldD1
  • Dehalogenase HldD1, encoding gene thereof and application of dehalogenase HldD1
  • Dehalogenase HldD1, encoding gene thereof and application of dehalogenase HldD1

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Embodiment 1

[0059] Embodiment 1, expression and activity determination of HldD1 protein

[0060] 1. Acquisition of the gene encoding the HldD1 protein

[0061] Using the genomic DNA of the new strain D1 isolated from deep-sea sediment samples in the Indian Ocean as a template, PCR amplification was performed with D1F and D1R primers:

[0062] D1F: 5'-AAGAAGGAGATATACATATGGCCATCGACGCCCTGCGCACC-3'

[0063] D1R: 5'-TCGAGTGCGGCCGCAAGCTTGCCGAGCCCGAACTTCTCCAG-3'

[0064] The PCR amplification product was recovered and cloned into the pET21a(+) vector (Novagen) to obtain the recombinant vector pET-D1, which was transformed into Escherichia coli BL21(DE3) and then sent for sequencing.

[0065] Sequencing results show that: the vector contains the nucleotides shown in sequence 1, and the gene in sequence 1 is named HldD1, the coding region of the gene is the 1-924th position of sequence 1, and the protein encoded by the gene is named HldD1, The amino acid sequence of HldD1 is shown in the 1-308 ...

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Abstract

The invention discloses dehalogenase HldD1, an encoding gene thereof and application of the dehalogenase HldD1. The invention provides a protein represented by a sequence 2 of a sequence list. Experiments prove that the dehalogenase HldD1 obtained through subjecting a gene sequence (sequence 1) disclosed by the invention to prokaryotic expression through Escherichia coli has high activity and has relatively high organic solvent stability separately under the treatment conditions of not higher than 20% of ethanol, not higher than 20% of acetonitrile and not higher than 40% of DMSO.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a dehalogenase HldD1 and its coding gene and application. Background technique [0002] Halogenated organic compounds are widely used in many fields. Among them, due to their extensive use in industrial enterprises and daily necessities, they have become an important class of environmental pollutants. These halogenated organics have continued to cause widespread environmental problems in the biosphere for many years due to their very stable chemical bonds. Halogenated organics are the most used and studied class of organics, mainly due to their relatively simple synthetic routes. However, due to the generally high stability of halogenated organic substances and their toxicity to microorganisms that may degrade them, the biodegradability of halogenated organic substances has been poor for many years. Excessive exposure to these halogenated pollutants can lead to serious ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/14C12N15/55C12N15/70C12N1/21C12P5/02C12R1/19
CPCY02A50/20
Inventor 董志扬张重雄王丽胡然戴欣
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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