Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof

A technology of cinnamyl alcohol dehydrogenase and coding gene, applied in the biological field, can solve the problems of complex process, low yield of extract and high cost

Inactive Publication Date: 2012-02-08
GRADUATE SCHOOL OF THE CHINESE ACAD OF SCI GSCAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biosynthesis of artemisinin is special, and it is only synthesized in specially differentiated cells—glandular secretory trichome (GST) on leaves and flower buds, which leads to the extremely low content of artemisinin in Artemisia annua , ab

Method used

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  • Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof
  • Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof
  • Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof

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Experimental program
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Embodiment 1

[0037] Embodiment 1, preparation and functional verification of enzyme

[0038] 1. Acquisition of the full-length cDNA sequence of the gene AaCAD encoding cinnamyl alcohol dehydrogenase from Artemisia annua

[0039] (1) Cloning of the 3' end sequence of the cinnamyl alcohol dehydrogenase gene AaCAD of Artemisia annua

[0040] The total RNA of the leaves of Artemisia annua (Artemisia annha L.) (the seeds of Artemisia annua were collected from Youyang, Chongqing, China) was extracted using the Universal Plant Total RNA Extraction Kit of Biotech Biotechnology Co., Ltd. The reverse transcription kit reverse-transcribes the total RNA into cDNA, and the primer used for reverse transcription is OligodT-adaptor 5′-CTG ATC TAG AGG TAC CGG ATC CAT ATT TTT TTT TTT TTTTTT-3′.

[0041] Primers were designed according to the ORF of the partial sequence of the cinnamyl alcohol dehydrogenase gene obtained from the Artemisia annua glandular trichome library, and the sequences of the specific ...

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Abstract

The invention discloses protein with cinnamyl alcohol dehydrogenase activity and a coding gene as well as application thereof. The protein with cinnamyl alcohol dehydrogenase activity provided by the invention is the protein shown as (a) or (b): (a) protein constituted by an amino acid sequence shown as a sequence 2 in a sequence table; and (b) protein derived from (a) by substituting and/or deleting and/or adding one or more amino acid residues for the amino acid sequence shown as the sequence 2 in the sequence table and with the cinnamyl alcohol dehydrogenase activity. By adopting the protein with the cinnamyl alcohol dehydrogenase activity provided by the invention, conversion of coniferyl aldehyde, sinapaldehyde and geranialdehyde into the form of alcohols thereof can be efficiently catalyzed, and conversion of precursor artemisinin aldehyde for artemisinin biosynthesis into artemisinin alcohol can also be catalyzed in an in-vitro enzymatic reaction system, and the later can be applied to adjusting and controlling an artemisinin biosynthesis process from a molecular level; and thus, a foundation is laid for producing a high-yield artemisinin strain.

Description

technical field [0001] The invention relates to a protein with cinnamyl alcohol dehydrogenase activity and its coding gene and application in the field of biotechnology. Background technique [0002] Lignin is the second most abundant organic matter in the world after cellulose. It is mainly a complex phenolic polymer formed by three kinds of lignin alcohol monomers, namely p-coumaroyl alcohol, coniferyl alcohol and sinapyl alcohol. As one of the main polymers in the cell wall, lignin is filled in the cellulose framework, which can enhance the mechanical strength of the plant and prevent the plant from lodging, which is conducive to the water transport of the tissue. At the same time, lignin and many related products have many functions in plant biotic or abiotic resistance, and can resist the invasion of adverse external environments. In addition, lignin is also very beneficial to human health. Lignin generally exists in the seeds of beans, wheat bran, cocoa, strawberries...

Claims

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Application Information

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IPC IPC(8): C12N9/04C12N15/53C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11C12P7/22C12P7/04C12P7/02
Inventor 王红李星马东明陈建林
Owner GRADUATE SCHOOL OF THE CHINESE ACAD OF SCI GSCAS
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