Recombinant Escherichia coli zjut-ho1 and its application in the preparation of biliverdin

Abstract

The invention discloses a recombinant Escherichia coli zjut-ho1 and its application in the preparation of biliverdin, which is preserved in the Guangdong Microbial Culture Collection Center, with a preservation number of GDMCC No: 60372 and a preservation date of May 18, 2018. Address: 5th Floor, Building 59, Compound, No. 100 Xianlie Middle Road, Guangzhou City, Zip Code: 510075. The recombinant Escherichia coli expresses HO-1 with intracellular activity, and the recombinant Escherichia coli can be used as the HO-1 enzyme source, and hemin is used as the substrate to prepare biliverdin by biotransformation, which improves the production of biliverdin. way. Compared with the existing similar technologies, the present invention expresses ho1 from prokaryotic organisms in Escherichia coli of prokaryotic organisms, which has the advantages of high HO-1 activity and short culture period, and uses hemin as a substrate to make The yield of biliverdin is greatly improved, which is 2.6 times that of the recombinant Escherichia coli whole cell synthesis method.

Description

(1) Technical field [0001] The invention relates to a gene recombinant Escherichia coli strain expressing heme oxygenase-1 derived from algae, and the application of the strain in preparing biliverdin by biotransforming heme. (2) Background technology [0002] Biliverdin (biliverdin, CAS: 114-25-0) is a linear tetrapyrrole ring substance obtained by hydrolysis of heme by heme oxygenase (hemeoxygenase-1, HO-1) (reaction formula see figure 1 ), named for its dark green color. In vertebrate cells, under the hydrolysis of HO-1, heme opens at the α-methylene bridge to form biliverdin IXα isomers. Therefore, biliverdin generally refers to biliverdin IXα. [0003] Biliverdin is not only an intermediate metabolite of the heme metabolic circulatory system, but more importantly, it can initiate a series of physiological effects such as anti-inflammation, anti-oxidation and immune regulation, such as improving liver function and reducing alanine aminotransferase , Alleviate the ische...

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Problems solved by technology

The extraction method is to extract biliverdin directly from salmon bile or emu eggshell, but the problem is that the source of raw materials is insufficient and it is difficult for industrial application; the chemical conversion method is prepared by dehydrogenating bilirubin, but there are two problems in this method: ①Bilirubin with high price is needed as a substrate, and the source of raw materials is difficult; ②Bilirubin is extracted from the bile of mammals, and needs to be under acidic conditions, which makes bilirubin form multiple isomers, resulting in the product bile The yield and purity of chlorophyll IXα are relatively low; there are also reports on the preparation of biliverdin by the chemical oxidation heme method, which also produces more isomers with a lower yield; A patent from cyanobacteria recombines the HO-1 gene from cyanobacteria into yeast cells, expresses HO-1, and then uses heme as a substrate to produce biliverdin through whole cell biotransformation. The advantage of this method is that there are few isomers , but the patent does not report the yield

It can be seen that there is currently a lack of economical and effective biliverdin preparation methods

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