Genetically engineered rhodobacter sphaeroides, preparation method thereof and farnesol production method

A technology of Rhodobacter sphaeroides and genetic engineering, applied in the field of Rhodobacter sphaeroides and the production of farnesol by using Rhodobacter sphaeroides, can solve the problems of less farnesol and low yield, and achieve the effect of simple method and easy extraction

Active Publication Date: 2017-09-01
FUJIAN FUKANG PHARMA
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  • Claims
  • Application Information

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Problems solved by technology

[0005] Chinese patent application CN103571763A discloses a strain of Saccharomyces cerevisiae screened from liquor grains. This Saccharomyces cerevisiae can synthesize various terpene alcohols from scratch. In addition to producing farnesol, the yeast can also synthe

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  • Genetically engineered rhodobacter sphaeroides, preparation method thereof and farnesol production method

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Example Embodiment

[0038] The preparation method of the genetically engineered Rhodobacter sphaeroides of the present invention includes the steps of constructing and transforming a recombinant plasmid and the steps of transforming the recombinant plasmid into Rhodobacter sphaeroides by bacterial conjugation.

[0039] In an exemplary embodiment, the step of constructing and transforming a recombinant plasmid includes:

[0040] The phosphatase PgpB1 gene was cloned from the genome of Yersinia pestis CO92; the pBBR1MCS2 vector was used as a template to clone the spliced ​​fragment of the pBBR1MCS2 vector, and the spliced ​​fragment was recovered; the PgpB gene and the spliced ​​fragment were mixed and reacted Then, transform E. coli T1 cells, pick positive clones, extract plasmid pBBR1MCS2-PgpB, transform it into host cells, pick positive strains, extract plasmids for verification, and obtain donor bacteria.

[0041] In an exemplary embodiment, the step of bacterial conjugation and transformation of the ...

Example Embodiment

[0053] Example 1

[0054] By heterologously expressing the Yersinia pestis phosphatase PgpB in the Rhodobacter sphaeroides 2.4.1 cell, the farnesyl ester accumulated in the MEP pathway of Rhodobacter sphaeroides can be converted into farnesol, thereby realizing the utilization of red sphaeroides. Bacteria to produce farnesol. Specific steps are as follows:

[0055] 1. Construct pBBR1MCS2-PgpB recombinant plasmid and transform E.coli S17-1 (donor bacteria):

[0056] The pBBR1MCS2-PgpB recombinant plasmid was constructed using Gibson Assembly (NEB Company, USA) seamless connection method. First, the genome of Yersinia pestis CO92 was extracted with a genome extraction kit (Quanshijin Company, Beijing). Using the genome as a template, the phosphatase PgpB1 gene was cloned from the genome using primers PgpB-F (SEQ ID NO: 2) and PgpB-R (SEQ ID NO: 3); using the pBBR1MCS2 vector as a template, primers pBBR1MCS2-F( SEQID NO: 4) and pBBR1MCS2-R (SEQ ID NO: 5) were cloned to obtain the sp...

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Abstract

The invention discloses genetically engineered rhodobacter sphaeroides, a preparation method thereof and a farnesol production method. A phosphatase PgpB gene from Yersinia pestis CO92 is transformed into rhodobacter sphaeroides and can be subjected to intracellular expression, and phosphatase PgpB with activity is generated, so that arnesyl pyrophosphate accumulated in the MEP pathway of rhodobacter sphaeroides can be directly catalyzed to generate farnesol.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rhodobacter sphaeroids and a method for producing farnesol by using the rhodobacter sphaeroids. Background technique [0002] Farnesol (Farnesol) is a kind of acyclic sesquiterpene alcohol, chemical formula C 15 h 26 O, present in trace amounts in the essential oils of Brazilian sandalwood, ambrette seed, acacia, citronella, lemongrass, musk, jasmine and neroli. For a long time, farnesol has been widely used as a fragrance ingredient in the cosmetics industry. In recent years, studies have found that farnesol also has various biological activities such as antibacterial, anti-inflammatory and anti-tumor. In addition, farnesol as a drug ingredient can significantly increase the susceptibility of various pathogenic microorganisms (such as Staphylococcus aureus, Candida albicans, Streptococcus mutans, Fusarium graminearum, etc.) to drugs. [0003] At present, the production and extr...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12P7/04C12R1/01
CPCC12N9/16C12N15/74C12P7/04
Inventor 祁峰黄建忠陈学端江贤章张明亮
Owner FUJIAN FUKANG PHARMA
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