Method for producing 5-glycyl ethylformic acid by using engineering bacterium

A technology of aminolevulinic acid and engineering bacteria, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of complex culture conditions, many by-products, and complicated steps, and achieves broad industrialization prospects. The effect of high controllability and simple process

Inactive Publication Date: 2007-09-26
ZHEJIANG UNIV
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Problems solved by technology

[0003] At present, the production of ALA mostly adopts the chemical synthesis method, which has cumbersome steps, many by-products, low yield and serious environmental pollution; while the biological mutagenesis method mainly ...

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  • Method for producing 5-glycyl ethylformic acid by using engineering bacterium

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Embodiment

[0018] The method for producing 5-aminolevulinic acid with engineering bacteria comprises the following steps:

[0019] 1. Use an inoculation needle to dip a small amount of bacterial liquid from the glycerol tube of CGMCC No.1939 engineering bacteria Rosetta (DE3)-pET28a-R.S.hemA, and add it to the chloramphenicol containing 30 μg / ml kanamycin and 34 μg / ml chloramphenicol. Streak on the LB medium plate and cultivate overnight in a 37°C oven;

[0020] 2. Pick a single clone and inoculate it in a 250ml shake flask containing 50ml LB medium at a rotation speed of 200rmp, and culture overnight at 37°C to obtain first-grade seeds;

[0021] 3. Take 1ml of primary seeds and inoculate them in a 500ml shake flask containing 100ml of fermentation medium, culture at 37°C and 200rpm for 3h to obtain secondary seeds. Wherein the initial fermentation medium is composed of 1% peptone, 0.5% yeast powder, 0.3% succinic acid, 0.2% glycine and 0.2% glucose in mass volume ratio, and the pH valu...

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Abstract

The invention discloses a method to prepare 5-amino levulinic acid with engineering bacteria, which comprises the following steps: 1) flat-activating engineering bacteria Rosetta(DE3)-pET28a-R.S.hemA with conserve number at CGMCC No.1939 through LB culture medium; 2) seeding the monoclonal on plate into shake flask of LB culture medium; overnight-culturing at 37 deg.c; getting the first grade seed; 3) seeding the first grade seed into shake flask of LB culture medium; getting the second grade seed; 4) seeding the second grade seed into fermentation tank with ferment culture medium; utilizing isopropyl-beta-D-sulfo galactose-glycoside to cool and evoke; culturing effluxion of time continually; proceeding supplement material culture.

Description

technical field [0001] The invention relates to a method for producing 5-aminolevulinic acid with engineering bacteria. Background technique [0002] 5-Aminolevulinic acid (5-aminolevulinic acid, ALA) widely exists in organisms, is the first compound in the biosynthesis pathway of porphyrin, and is also used in the synthesis of heme, chlorophyll, vitamin B 12 common precursor of tetrapyrrole compounds. As a photodynamic agent (photodynamic agent, PDT), ALA has a wide range of uses in the fields of agrochemicals and medicine. In the field of agriculture, ALA has many functions such as weeding, insecticide, increasing plant stress resistance and promoting plant growth, etc., and is easy to degrade without residue, non-toxic to humans and animals, and has become a very promising pollution-free green agrochemical. In the field of medicine, ALA has the effect of selectively killing cancer cells. It is called the second-generation photodynamic medicine (photodynamic medicine). ...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/01
Inventor 林建平傅维琦岑沛霖
Owner ZHEJIANG UNIV
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