TH76151A - Biochemical synthesis of 1,4-Butane Diamine - Google Patents

Abstract

DC60 (10/10/40) Invention involves a process for the biosynthesis of 1,4-butanediamine in microorganisms with increased activity levels of ornithene decarboxylase. (Increased ODC activity) compared to the natural level of ornithene carboxylase activity, where the increase in ODC activity was Comes by way of expressing more than usual in China Where the efficient ornithiniccarboxylase code is processed, enhanced decoding and / or decoding, and where 1,4-butane dioxide generated in microorganisms is obtained The release into the fermented and collected from the fermented water in popular detailing, the enzyme-enhanced activity was obtained by acting. Excessive output of (i) Chinese speA that composes Argenene decarboxylase and Chinese speB that encodes the actinase code, or (ii) Chinese speA that encodes Argenine code. Decarboxylase and China aguA coding Agmatin, iminohydrolase and Chinese code-coded aguB N-carbamoil puretresene amidohydrolase, and may also select Chinese speB that encodes actinase code. The invention also involves plasmid and host vectors with increased activity. Of the activity of one or more enzymes, as mentioned Invention involves a process for biosynthesis of 1,4-butanediamine in microorganisms with increased activity levels of ornithene decarboxylase. (Increased ODC activity) compared to the natural level of ornithene carboxylase activity, where the increase in ODC activity was Comes by way of expressing more than usual in China Where the efficient ornithiniccarboxylase code is processed, enhanced decoding and / or decoding, and where 1,4-butane dioxide generated in microorganisms is obtained The release into the fermented and collected from the fermented water in popular detailing, the enzyme-enhanced activity was obtained by acting. Excessive output of (i) Chinese speA that composes Argenene decarboxylase and Chinese speB that encodes the actinase code, or (ii) Chinese speA that encodes Argenine code. Decarboxylase and China aguA coding Agmatin, iminohydrolase, and Chinese aguB coded N-carbamoil puretresene amidohydrolase, and may also select Chinese speB that encodes actinase code. The invention also involves plasmid and host vectors with increased activity. Of the activity of one or more enzymes, as mentioned

Claims (21)

1. The process for the biosynthesis of 1,4-bitendiamine in microorganisms with increased levels of ornithine decarboxylase activity (increased ODC activity) compared to natural levels of ornithine decarboxylase activity is characterized by the increase in ornithine decarboxylase activity through the overexpression of ornithine decarboxylase encoding genes, resulting in increased transcription and/or translation efficiency, and the generated 1,4-bitendiamine is released into the fermentation broth and collected from the broth. 2. The procedure under Claim 1, whereby the enhanced decoding and/or translation efficiency is achieved by the use of a commonly controlled, strong promoter or a strong induced promoter. 3. The procedure under claim 2, whereby the enhanced decoding and/or translation efficiency is achieved by using a potent promoter induced by isopropyl-beta-D-thiogalactoside (IPTG). 4. The procedure under Claim 1, whereby the enhanced decoding and/or translation efficiency is achieved by using a promoter selected from a group comprising the promoters T7, T5, ptac, and plac. 5. Procedure according to claim 1, where the ornithine decarboxylase encoding gene has a ribosome binding site (RBS) located to the left of the gene's encoding region, and the RBS is adapted to better recognize the RNA template by ribosomes. 6. Procedure under Claim 1, where the gene encoding ornithyrene decarboxylase is ornithyrene decarboxylase speF or speC (each listed in E.C.4.1.1.17). 7. Procedure under Claim 6, where the gene processing the overexpressed ornithine decarboxylase is the ornithine decarboxylase speF gene. 8. Procedure pursuant to claim 6, where the ornithine decarboxylase encoding gene that is overexpressed is the ornithine decarboxylase sprF or speC gene from one of the selected genera from the group comprising Escherichia, Shigella, Salmonella, Yersinea, and Shewanella. 9. Procedure pursuant to claim 8, where the ornithine decarboxylase encoding gene that is overexpressed is an ornithine decarboxylase gene from one of selected species from a group comprising Escherichia coli, Shigella flexneri, Salmonella typhimutium, Yersinea pestis, and Shewanella oneidensis. 10. Procedure pursuant to claim 9, where the gene encoding the overexpressed ornithine decarboxylase is speF, derived from one of selected species from a group comprising Escherichia coli, Salmonella typhimurium, and Shewanella oneidensis. 11. Procedure according to Claim 1, whereby in addition to the increased activity of ODC, the increased activity of at least two other enzymes is also obtained by the overexpression of (i) the speA gene encoding arginine decarboxylase (classified in E.C.4.1.1.19) and the speB gene encoding agmatinase (classified in E.C.3.5.3.11; also referred to as the gene encoding agmatin urea hydrolase), or (ii) the speA gene encoding arginine decarboxylase (classified in E.C.4.1.1.19) and the aguA gene encoding agmatin iminohydrase (classified in E.C.3.5.3.12; also referred to as the gene encoding agmatin deiminase) and the aguB gene encoding N-carbamoyl putrasine amide hydrolase (classified in E.C.3.5.1.53) and may also select the speB gene that processes the agmatinae code (classified in E.C.3.5.3.11). 12. Procedure pursuant to claim 11, where the gene encoding the overexpressed arginine decarboxylase speA is derived from one of the selected genera from the group comprising Escherichia, Shigella, Salmonella, Yersinia, Pasteurella, and Neisseria. 13. Procedure pursuant to claim 12, where the encoding gene for the overexpressed arginine decarboxylase is an arginine decarboxylase speA gene derived from one of a selected species from a group comprising Escherichia coli, Shigella flexneri, Salmonella enterica, Yersinia pestis, Pasteurella multocida, and Neisseria meningitidis. 14. Procedure pursuant to claim 13, whereby the gene encoding the overexpressed acmatinae is an acmatinae speB genera derived from one of the selected genera from the group comprising Escherichia, Salmonella, Proteus, Photorhabdus, Vibrio, and Neisseria. 15. Procedure pursuant to claim 14, where the gene encoding the overexpressed acmatinae is an acmatinae speB genera derived from one of the selected genera from the group comprising Escherichia coli, Salmonella enterica, Proteus mirabilis, Photorhabdus luminescens, Vibrio cholerae, and Neisseria meningitidis. 16. Procedure pursuant to claim 15, where the encoding gene for acmateine imidohydrase and/or the encoding gene for N-carbamoyl purestracene amide hydrolase is the gene aguA acmateine imidohydrase and/or the gene aguB N-carbamoyl purestracene amide hydrolase derived from one of the selected genera from the group comprising Pseudomonas, Streptococcus, Streptomyces, Azotobacter, Arabidopsis, Novosphingobium, and Bacillus. 17. Procedure pursuant to claim 16, where the encoding gene for acmateine imidohydrase and/or the encoding gene for N-carbamoyl purestracene amide hydrolase is the gene aguA acmateine imidohydrase and/or the gene aguB N-carbamoyl purestracene amide hydrolase derived from one of the selected genera from the group comprising Pseudomonas aeruginosa, Streptococcus mutans, Streptomyces avermitilis, Azotobacter vinelandii, Arabidopsis thaliana, Novosphingobium aromaticivorans, and Bacillus cereus. 18. A procedure pursuant to one of the claims 1 through 17 is performed while ensuring that intracellular levels of ornithine are increased. 19. A procedure under any one of the claims in claims 1 through 17, whereby the procedure is performed on selected host organisms from the group comprising Saccharomyces sp., Bacillus sp., Corynebacterium sp., Escherichia sp., and Pichia sp. 20. A procedure pursuant to one of the claims in claims 1 through 17, whereby the procedure was performed in selected host organisms from the group comprising Saccharomyces cerevisiae, Corynebacterium sp., and Escherichia sp., and whereby in addition to the level of ornithine decarboxylase activity at least relative to the level of arginine decarboxylase activity, together with agmatinae and/or agmatine iminohydrase, and N-carbamoyl purtresine aminohydrase, was added. 21. Vectors, plasmids, and hosts containing at a level of increased activity of one or more enzymes as described in any one of the claims of Reputation 1-20.