Microorganism and process for the preparation of L-methionine
The present invention relates to microorganisms and processes for the efficient preparation of L-amino acids such as L-methionine. In particular, the present invention relates to microorganisms and processes in which the formation and/or accumulation of homolanthionine in the methionine pathway is reduced and/or prevented.
1. A process for the preparation of L-methionine, which comprises the following steps:
cultivating and/or fermenting a microorganism, wherein said microorganism is a Corynebacterium glutamicum cell that has its endogenous mcrB and metB genes disrupted by a deletion or an insertion, wherein said microorganism produces or overproduces L-methionine and in which the formation and/or accumulation of homolanthionine in the methionine pathway is reduced and/or prevented, wherein the production of an endogenous transcriptional regulator protein (McbR) is reduced compared to that of the corresponding wild-type microorganism, and the microorganism
(a) has introduced therein a heterologous gene coding for a cystathionine-β-lyase (MetC) mutant which is capable of efficiently converting homolanthionine into homocysteine,
(b) has introduced therein a heterologous gene coding for a cystathionine-γ-synthase (MetB) mutant which is capable of efficiently converting O-acetyl-homoserine and cysteine into cystathione and which is not capable of converting O-acetyl-homoserine and homocysteine into homolanthionine, or
(c) produces more of a protein selected from the group consisting of a protein having the activity of cob(I)alamin dependent methionine synthase I (MetH), and cob(I)alamin independent methionine synthase II (MetE) than the corresponding wild-type microorganism; and
isolating L-methionine.
2. The process of claim 1 , wherein the production of an endogenous cystathionine-γ-synthase (MetB) is reduced compared to the corresponding wild-type microorganism.
3. The process of claim 2 , wherein an endogenous gene which codes for cystathionine-γ-synthase (MetB) is disrupted or eliminated.
4. The process of claim 3 , wherein the disrupted endogenous gene results in the expression of a non-functional cystathionine-γ-synthase (MetB) protein in the cultivated microorganism.
5. The process of claim 1 , wherein an endogenous gene which codes for transcriptional regulator protein (McbR) is disrupted and/or eliminated.
6. The process of claim 5 , wherein the disrupted endogenous gene results in the expression of a non-functional transcriptional regulator protein (McbR) protein.
7. The process of claim 1 , wherein expression of a protein selected from the group consisting of a protein having the activity of cob(I)alamin dependent methionine synthase I (MetH), and cob(I)alamin independent methionine synthase II (MetE) is increased as compared to the corresponding wild-type microorganism.
8. The process of claim 1 , wherein L-methionine is concentrated in the medium or in the microorganism.
9. A process for the preparation of a L-methionine containing animal feedstuffs additive from fermentation broths, comprising the following steps:
cultivating and/or fermenting a microorganism, wherein said microorganism is a Corynebacterium glutamicum cell that has its endogenous mcrB and metB genes disrupted by a deletion or an insertion, wherein said microorganism produces or overproduces L-methionine and in which the formation and/or accumulation of homolanthionine in the methionine pathway is reduced and/or prevented;
wherein the microorganism
(a) has introduced therein a heterologous gene coding for a cystathionine-β-lyase (MetC) mutant which is capable of efficiently converting homolanthionine into homocysteine,
(b) has introduced therein a heterologous gene coding for a cystathionine-γ-synthase (MetB) mutant which is capable of efficiently converting O-acetyl-homoserine and cysteine into cystathione and which is not capable of converting O-acetyl-homoserine and homocysteine into homolanthionine, or
(c) produces more of a protein selected from the group consisting of a protein having the activity of cob(I)alamin dependent methionine synthase I (MetH), and cob(I)alamin independent methionine synthase II (MetE) than the corresponding wild-type microorganism;
removing water from the L-methionine-containing fermentation broth; removing an amount of 0 to 100 wt. % of the biomass formed during fermentation; and
drying the fermentation broth to obtain the animal feedstuffs additive in powder or granule form.