TH64114B - The use of dimethyl disulfide for methionine production in microorganisms. - Google Patents

Abstract

DC60 (15/06/16) This invention characterizes the improved process. and methionine-producing organisms. Fabrication has shown that deltametF or deltametE deltametH organisms, for example, mutants of C. glutamicum or E. coli, can Use a methyl enclosed sulfide source such as dimethyl disulfide (DMDS) as a source of both sulfur and methyl groups by bypassing the activity requirements of MetH/MetE and MetF and want to reduce sulfate Also described in the patent are information related to MetY (also referred to as MetZ), an enzyme that incorporates methyl capd sulfide sources such as DMDS into methionine. Neene, deltametF, deltametB of C. glutamicum can use a methyl capd sulfide source such as DMDS as a source of both sulfur and methyl groups. Methionine production by prototropic organisms that have been over-engineered O-acetyl-homoserine It has been enhanced by adding a methyl capd sulfide source such as DMDS. Revised Invention Summary 15/06/16 This invention characterizes the improved process. and organisms for the production of methionine. Inventions have shown that organisms (formula)metF or organisms (formula)metE (formula)metH, for example, mutants of C. glutamicum or E. coli, can Use a methyl capd disulfide source such as dimethyl disulfide (DMDS) as a source of both sulfur and methyl groups by bypassing the activity requirements of MetH/MetE and MetF and want to reduce sulfate Also described in the patent are information related to MetY (also referred to as MetZ), an enzyme that incorporates methyl capd sulfide sources such as DMDS into methionine. The nine strain (formula)metF (formula)metB of C. glutamicum can use a methyl capd sulfide source such as DMDS as a source of both sulfur and methyl groups. Methionine production by prototropic organisms that have been over-engineered O-acetyl-homoserine It is enhanced by adding a methyl capd sulfide source such as DMDS ------------------------------- --------------------------------------- This invention characterizes the improved process. and organisms for the production of methionine. Inventions have shown that organisms (formula)metF or organisms (formula)metE (formula)metH, for example, mutants of C. glutamicum or E. coli, can Use a methyl enclosed sulfide source such as dimethyl disulfide (DMDS) as a source of both sulfur and methyl groups. By bypassing the activity requirements of MetH/MetE and MetF and the need to reduce sulfate. Also described in the patent is information related to MetY (also referred to as MetZ), an enzyme integrated into the source. Methyl-enclosed sulfides, such as DMDS, enter methionine. The metF (formula) metB species of C. glutamicum can use a methyl-enclosed sulfide source such as DMDS as the source. of both the sulfur and methyl groups. Moreover, the production of metheonine The engineered prototropic organisms that overproduce O-acetyl-homocerine were enhanced by the addition of a methyl enclosed sulfide source such as DMDS.

Claims (6)

Disclaimer (all) which will not appear on the announcement page: EDIT 15/06/16 1. Methionine production method, which consists of culturing methionine-producing microorganisms. With Methylcapide sulfide (meathyl capped sulfde compound) is included, so that methionine is produced, where methylcapide sulfide is H3C- (S ) n-CH3 and n are 2-50 2. Methionine production method, combining methionine-producing microorganisms with dimethyl disulfide (DMDS ), So that methionine was produced 3. Method of 1st or 2nd duty, with DMDS of 0.02% or higher in culture. 1 or 2, with DMDS 0.06% or higher in culture 5. Methods of methionine production, consisting of methionine-producing microorganisms cultured with a system. Transport of slow-release dimethyl disulfide (DMDS), in a manner in which methionine is produced. 6. Method of claim 5, where the slow-release DMDS transport system is composed of methionine. Combined with liquids that are incompatible with water, but dissolved in DMDS 7. Method of claim 6, where the slow-release DMDS transport system consists of a liquid of choice from a group containing: oil. Animal, mineral oil, chemical oil, vegetable oil, synthetic oil 8) Method of claim 5, whereby the transport system is used as a means of transporting the organic solvent, chloro-carbon, fluoro-carbon, chloro-fluoro-carbon or its combination. Slow release DMDS is the regulated slow feed of the DMDS 9. The method of holding rights 5, where the slow release DMDS transport system is the flow or diffusion of the DMDS through the permeable membrane. Continued DMDS 1 0. Claim Method 5, where the slow release DMDS transport system incorporates DMDS input in the gas state 1 1. Claim Method 10, where DMDS in State Gas is generated by evaporating or boiling liquid DMDS 1 2. Method of Proposition 10, where gas-state DMDS is generated by air bubbles or oxygen through liquid DMDS 1 3. Method of claim 2 and any one, where methionine-producing microorganisms are in the genus Corynebacterium 1 4. Method of claim 2 and any one, where the microorganisms producing methionine is Corynebacterium glutamicum 1 5. Method of either claim 2 and 5, where Methionine-producing microorganisms were selected from The group consists of Gram-Negative, Bacteri-Positive, Yeast and Archea 1 6. Method of one of the 2nd and 5th Clause, where methionine-producing microorganisms contain the enzyme that Biosynthesis of methionine was at least one type of deregulate 1 7. Method of claim 16, whereby the microorganisms have O-acetyl-homosyran Sulfhydrylase or O-Sulfinil-Hymoserene Dehydrated sulfhydrylase 1 8. Method of either Clause 2 and 5, where methionine-producing microorganisms contain the enzyme that Biosynthesis of at least two biodegradable methionine delegated 1 9. Method of claim 18, whereby the microorganisms contain homosirine. Acetyltransferase or homocerene, succinyl transferase, de-regulated and homocerene dehydrogenase Derekulet 2 0. Method of claim 18, whereby such microorganisms have O-acetyl-homosyrine Sulfhydrylase Dereggulate and homosyrene acetyltransferase dereggulate or O-succinil-homosyrine De-regulated sulfhydrylate and de-regulated homocyrene succinyl transferase 2 1. Methi production method. Onene, which consists of microorganism culture with a synthetic pathway. Biodegradable methionine with dimethyl disulfide (DMDS), so that methionine Was produced 2 2. Method of claim 21 where the microorganisms belong to the genus Corynebacterium 2 3. Method of claim 21, where the microorganism is Corynebacterrium glutamicum 2 4. Method of claim 21, where the microbes belong to the genus Escherichaia 2 5. Method of claim 21, Where the microorganisms were selected from a group consisting of: Gram-Negative Bacteria, Gram-Positive Bacteria, Yeast and Archea 2 6. Method of claim 21, whereby the microorganism had O-acetyl-homosyrine Sulfhydrylase or O-succinil-homosyrine Defined sulfhydrylase 2 7. Method of claim 21, whereby the microorganisms contain homosirine Acetyltransferase or homocerene, sulfinyl transferase, de-regulated, and corrected homoseri dehydrogenase Derekulet 2 8. Method of claim 21, whereby such microorganisms have O-acetyl-homosyran sulfhydrylase Dereggulate and homosyrene acetyltransferase dereggulate or O-succinil-homosyrine Dehydrated sulfhydrylate And homocyrin succinyl transferase 2 9. The method of claim 21, which also includes the process of methio extraction. Nene 3 0. Pre-combinant microorganisms that produce methionine with dimethyl disulfide (DMDS), such microorganisms. Biological methionine biosynthesis pathway is derekulate 3 1. The microbes of claim 30 belong to the genus Corynebacterium 3 2. The microbes of claim 31, which is Corynebacterium glutamicum 3 3. The microbe of claim 30 is in the genus Escherichia --------------------------------------- --------------------------------- 1. Method of methionine production that consists of cultivating Cultivate methionine-producing microorganisms When methyl sulfide compounds such as methionine are produced 2. Method of claim 1 where methyl sulfide compounds are selectable from the group. that Contains dimethyl disulfide (DMDS), dimethyl trisulfide, dimethyl tetrosulfide, and a molecular weight sulfide polymer. Higher Its ends are closed with methyl groups. 3. Method of methionine production consists of microorganisms that produce methionine in the presence of dimethyl disulfide (DMDS). ) Until methionine 4. Any of the 1-3 Claim Method, where DMDS is 0.02% or higher in culture 5. Any of the 1-3 Claim Method by The DMDS was present at 0.06% or higher in culture. 6. Methionine production method consists of microorganism culture that produces methionine when available. Slow-release dimethyl disulfide (DMDS) transport system Until methionine was produced. 7. Method according to claim 6, where the slow-release DMDS transport system was Amberlite TM XAD4. Discharge DMDS at 0.1% or higher in culture 9. Method according to claim 7, where the slow-release DMDS transport system releases DMDS of 0.3% or higher in culture 1 0. Privilege 6 where the slow-release DMDS transport is composed of a liquid that is incompatible with water but dissolves in DMDS 1. 1. Method according to claim 10, where the slow-release DMDS transport system integrates. With selectable liquids from animal oil, mineral oil, chemical oil, vegetable oil, synthetic oil, organic solvent, chloro-carbon, fluorocarbon, chloro-fluoro- Carbon or its combination 1 2. Method according to claim 6, whereby the slow-release DMDS transport system is the slow feeding of the regulated DMDS 1 3. The method of claim 6, whereby the transport system Slowly releasing DMDS This is the flow or diffusion of DMDS through a permeable membrane per DMDS 1 4. Method according to claim 6, where the slow-release DMDS transport system consists of a DMDS feeding in the gas state 1 5. Method according to claim 14, where the gas-state DMDS is generated by evaporating or boiling the liquid DMDS 1 6. Method according to claim 14, wherein the gas-state DMDS is generated by supplying Air bubbles or oxygen through liquid DMDS 1 7. Any of the methods in Claim 1, 2, 3 or 6. Where methionine-producing microorganisms are in the genus Corynebacterium 1 8. Any method according to claim 1, 2, 3 or 6. Where methionine-producing microorganisms are in the genus Corynebacterium glutamicum 1 9. Any method according to claim 1, 2, 3 or 6. Where methionine-producing microorganisms are selectable From a group consisting of gram-negative, gram-positive, yeast and archaea 2 0. Any method according to claim 1, 2, 3 or 6. Where the methionine-producing microorganisms have enzymes The biological production of methionine was uncontrolled. 2 1. Method of claim 20, whereby the microorganisms have O-acetyl-homosyrine sulfhydrylase or O-succinil-homosyrine sulfhydrylase That were left uncontrolled 2 2. one of the methods of claim 1, 2, 3 or 6 Where the methionine-producing microorganisms have enzymes 2 3. Method according to claim 22, whereby the microorganisms contain homocyrene acetyltransfer. Race or homosyrine succinil transferase Out of control And made homocerene dehydrogenase Uncontrolled 2 4. Method according to claim 22, whereby such microorganisms have O-acetyl-homosyrine sulfhydrylase Out of control And homocerene acetyltransferase Uncontrolled or O-succinil-homoserene sulfhydrylase Out of control And homosyrine succinil Uncontrolled transferase 2 5. Methionine production method comprising of microorganism culture with synthetic pathway. Biological methionine That was uncontrolled when dimethyl disulfide (DMDS) was present until methionine was produced. 2. 6. Method according to claim 25, where the microorganisms belong to the genus. Corynebacterium 2 7. Method according to claim 25 where the microorganisms belong to the genus. Corynebacterium glutamicum 2 8. Method according to claim 25, where the microorganisms belong to the genus Escherichia 2 9. Method according to claim 25 where the microorganisms can be selected from the group containing gram-negative, gram-positive, yeast and Archia 3 0. Method according to claim 25, whereby such microorganisms have O-acetyl-homosyrine sulfhydrylase or O-succinil-homosyrine sulfhydrylase 3 1. Method according to claim 25, whereby the microorganisms contain homocyrene, acetyltransferase or homocerene succinyltrans. Sferes Out of control And made homocerene dehydrogenase Uncontrolled 3 2. Method according to claim 25 whereby the microorganisms have O-acetyl-homosyrine sulfhydrylase Out of control And homocerene acetyltransferase That were made without the control or O-succinil-homoserene sulfhydrylase Out of control And uncontrolled homocyrin succinyl transferase 3 3. Method according to claim 25, which also includes the methionine separation phase 3 4 Methionine-containing constituents isolated under claim 33 3. 5. Products containing methionine synthesized by any of the above claims 3 6. Microbial Recombinant in the genus Corynebacterium For the production of methionine In the presence of dimethyl disulfide (DMDS), the microorganisms have an uncontrolled pathway for biological methionine synthesis. 3 7. Recombinant microorganisms belonging to the genus. Corynebacterium glutamicum for methionine production. In the presence of dimethyl disulfide (DMDS), the microorganisms have a biologically modified pathway of methionine synthesis. Uncontrolled 3 8. Recombinant Escherichia microorganisms for methionine production. In the presence of dimethyl disulfide (DMDS), the microorganism has an uncontrolled pathway for biosynthesis of methionine. 3 9. Methods for improving DMDS utilization for methio production. Neen that consists of selection Oxotroph methionine that grows or grows faster on low-methionine-deficient diets but contains DMDS 4.0. Microorganisms obtained from the selection method of claim 39 or the offspring of microorganisms. Such 4 1. China isolated at the microbe derivative obtained by the selection method of claim 39 4. 2. Method for identifying coded mutant alleles O-acetylhydrylase or O-succinyl homoserene sulfhydrylase A) exposure to DMDS-dependent microorganisms and coded plasmids. O-acetylhydrylase or O-succinyl homoserene sulfhydrylase B) Selection of mutant mutants of the organisms that are resistant to methionine-free diets containing methionine analogs that inhibit the growth of such organisms. C) the separation of the mutant variant. Where the resistant phenotypes are coded by And d) DNA sequencing of the relative parts of the plasmid to indicate the presence of mutant plasmids. The order has been changed in the code area for O-acetylhydrylase, or O-succinyl homoseri, sulfhydrase until the mutated alleles that are processed Code O-acetylhydrylase or O-succinyl homoseri sulfhydrylase Resistance to feed inhibition Methionine was identified 4. 3.Method according to claim 42, where the organism lacks methionine synthase (MetE and / or MetH) or methyl tetrahydrophyleductase (MetF). And the aforementioned methionine-free food contains dimethyl disulfide (DMDS) 4 4. Method of claim 42 or 43, whereby the organism is a species of Corynebacterium glutamicum 4 5. Method of claim 42 or 43, where the organism is Escherichia coli 4 species. 6. Enzyme O-acetylhydrylase or O-succinyl homosyrene sulfhydrylase Mutations isolated by means of claim 42