Production of riboflavin
AssigneeDsm IP Assets B.V.
InventorsMartin Lehmann, Hans-Peter Hohmann, Dietmar Laudert, Michael Hans
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Abstract
The present invention provides an improved biotechnological production of riboflavin (also referred herein as vitamin B2) through modification in the operon containing the riboflavin biosynthetic genes (rib operon), in particular modifications of/in the leader sequences (rib leader) upstream of the corresponding riboflavin biosynthetic genes (rib operon). Furthermore, the present invention relates to genetically engineered microorganisms carrying said modified sequences, processes to generate said modified sequences/microorganisms and the use thereof for production of riboflavin.
Claims14
A mutated Bacillus subtilis rib leader sequence of the riboflavin biosynthesis operon operatively linked to at least one downstream rib gene,
- wherein said mutated Bacillus subtilis rib leader sequence does not comprise a Bacillus subtilis rib leader terminator sequence and does comprise at least one ribO mutation,
- wherein said Bacillus subtilis rib leader terminator sequence consists of a contiguous sequence of nucleotides 231-263 of SEQ ID NO:
- 42, and said at least one ribO mutation is selected from the group consisting of a mutation of one of the following positions of SEQ ID NO:42:
- T31, G39, G40, G41, C55, C85, C86, G88, C93, A116, G121 and C128.
A mutated Bacillus subtilis rib leader sequence of the riboflavin biosynthesis operon operatively linked to at least one downstream rib gene and a constitutive promoter, wherein said mutated Bacillus subtilis rib leader sequence operably linked to a constitutive promoter comprises the nucleotide sequence of SEQ ID NO:67, 68, 69, 70 or 71.
A mutated Bacillus subtilis rib leader sequence of the riboflavin biosynthesis operon operatively linked to at least one downstream rib gene,
- wherein said mutated Bacillus subtilis rib leader sequence does not comprise a Bacillus subtilis rib leader terminator sequence and does comprise at least one ribO mutation,
- wherein said Bacillus subtilis rib leader terminator sequence consists of a contiguous sequence of nucleotides 231-263 of SEQ ID NO:42, and said at least one ribO mutation is selected from the group consisting of a mutation of one of the following positions of SEQ ID NO:42:
- T31, G39, G40, G41, C55, C85, C86, G88, C93, A116, G121 and C128, and
- wherein said mutated rib leader sequence leads to relaxation or reduction of a repressive effect of a non-modified Bacillus subtilis rib leader sequence on expression of a downstream rib gene when said mutated rib leader sequence is expressed in a riboflavin biosynthesis operon as compared to expression of said non-modified Bacillus subtilis rib leader sequence in the riboflavin biosynthesis operon.
A mutated Bacillus subtilis rib leader sequence of the riboflavin biosynthesis operon operatively linked to at least one downstream rib gene,
- wherein said mutated Bacillus subtilis rib leader sequence does not comprise a Bacillus subtilis rib leader terminator sequence and does comprise at least one ribO mutation,
- wherein said Bacillus subtilis rib leader terminator sequence consists of a contiguous sequence of nucleotides 231-263 of SEQ ID NO:42, and said at least one ribO mutation is selected from the group consisting of a mutation of one of the following positions of SEQ ID NO:42:
- G39, G40, G41, C85 and G121.
Dependencies are inferred from the claim text.