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Astaxanthin-producing engineering bacterium based on dunaliella salina metabolic pathway and adonis aestivalis CBFD and HBFD and construction and application of astaxanthin-producing engineering bacterium

A technology of Dunaliella salina and Calendula officinalis, applied in genetic engineering, microorganism-based methods, enzymes, etc., can solve problems such as no astaxanthin production

Active Publication Date: 2020-11-10
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Dunaliella salina is a green alga with high production of β-carotene, which is the precursor of astaxanthin synthesis, but there is no related report on the production of astaxanthin in Dunaliella salina

Method used

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  • Astaxanthin-producing engineering bacterium based on dunaliella salina metabolic pathway and adonis aestivalis CBFD and HBFD and construction and application of astaxanthin-producing engineering bacterium
  • Astaxanthin-producing engineering bacterium based on dunaliella salina metabolic pathway and adonis aestivalis CBFD and HBFD and construction and application of astaxanthin-producing engineering bacterium
  • Astaxanthin-producing engineering bacterium based on dunaliella salina metabolic pathway and adonis aestivalis CBFD and HBFD and construction and application of astaxanthin-producing engineering bacterium

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Embodiment 1

[0044] 1. Preparation of Dunaliella salina culture solution

[0045] Prepare salina culture fluid without sodium chloride, the formula of the culture fluid is as follows: NaNO 3 0.420g / L, NaH 2 PO 4 2H 2 O 0.156g / L, NaHCO 3 0.840g / L, KCl 0.074g / L, MgSO 4 ·7H 2 O 1.230g / L, CaCl 2 2H 2 O0.044g / L, 0.1% Fe-EDTA solution 0.5mL / L. Add 1 mL / L of A5 trace element solution to the culture medium to adjust the pH to 7.5. Wherein, Fe-EDTA liquid composition is as follows: Na 2 EDTA 0.189g / L, FeCl 3 ·6H 2 O 0.244g / L; A5 trace element solution composition is as follows: H 3 BO 3 2.86g / L, MnCl 2 4H 2 O 1.81g / L, ZnSO 4 ·7H 2 O 0.22g / L, CuSO 4 ·5H 2 O 0.08g / L, (NH 4 ) 6 Mo 7 o 24 4H 2 O 0.04g / L.

[0046] 2. Construction of Dunaliella salina astaxanthin expression vector

[0047] The pCR2.1 vector sequence, the Dbgapdhpro promoter sequence, the RBS2 sequence (chloroplast localization peptide), the CBFD gene sequence of Calendula officinalis, the HBFD gene sequence o...

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Abstract

The invention discloses an astaxanthin-producing engineering bacterium based on a dunaliella salina metabolic pathway and adonis aestivalis CBFD and HBFD as well as construction and application of theastaxanthin-producing engineering bacterium. Dunaliella salina is adopted as a material, and exogenous genes adonis aestivalis astaxanthin are inserted to synthesize related genes CBFD and HBFD, so that the CBFD and the HBFD can be stably expressed in recombinant algae cells, conversion of the dunaliella salina from beta carotene to astaxanthin is realized, and accumulation of the astaxanthin inthe dunaliella salina is further realized. A new way is opened up for the production of natural astaxanthin.

Description

technical field [0001] The invention relates to the field of molecular cloning, in particular to an astaxanthin-producing engineering bacterium based on the metabolic pathway of Dunaliella salina and CBFD and HBFD of Calendula philodendron and its construction and application. Background technique [0002] Improving the biosynthesis of astaxanthin through metabolic engineering has gradually become a research hotspot in recent years. Since the main synthetic pathways and metabolic processes of astaxanthin in vivo have basically been described in detail, and genes encoding different enzymes in the astaxanthin synthesis route have been cloned from cells of bacteria, plants, algae and fungi, this It provides a basis for constructing engineering strains and optimizing astaxanthin synthetic metabolic network by using metabolic engineering technology. Haematococcus pluvialis or Phaffia rhodozyme, which can synthesize astaxanthin itself, can be used as receptors for metabolic engin...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/13C12N15/82C12P23/00C12R1/89
CPCC12N9/0071C12N9/00C12N15/8206C12P23/00
Inventor 姜建国陈浩宏谢红梁明华何羽婧
Owner SOUTH CHINA UNIV OF TECH