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isolated mutants of coryneform bacteria, isolated polynucleotide, vector, recombinant microorganism, and process for producing l-lysine
isolated mutants of coryneform bacteria, isolated polynucleotide, vector, recombinant microorganism, and process for producing l-lysine
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机译：棒状细菌的分离突变体，分离的多核苷酸，载体，重组微生物以及生产l-赖氨酸的方法
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摘要
Coryneform bacterium mutant comprises a gene coding for a 2-methylcitrate dehydratase polypeptide (I) with a proteinogenic amino acid other than L-proline at position 272. Independent claims are also included for: (1) coryneform bacterium mutant produced by treating a coryneform bacterium with a mutagenic agent, isolating and multiplying the resulting mutants, preparing nucleic acids from the mutants, producing a nucleic acid molecule from the nucleic acids by PCR using a primer comprising at least 15 consecutive nucleotides from the sequence between positions 1 and 1563 of SEQ ID NO:3 or SEQ ID NO:7 (not given in the specification) and a primer comprising at least 15 consecutive nucleotides from the complementary sequence between positions 3000 and 1567 of SEQ ID NO:3 or SEQ ID NO:7, sequencing the nucleic acid molecule and determining the encoded amino acid sequence, optionally comparing the sequence with SEQ ID NO:6 (not given in the specification), and identifying a mutant containing a polynucleotide coding for a polypeptide with a proteinogenic amino acid other than L-proline at position 272; (2) isolated polynucleotide (II) coding for (I); (3) isolated polynucleotide coding for an amino acid sequence corresponding to positions 19-57 of SEQ ID NO:2 (not given in the specification) with a proteinogenic amino acid other than L-proline at position 272; (4) producing a recombinant coryneform bacterium by transferring (II) into a coryneform bacterium so that the bacterium's 2-methylcitrate dehydratase gene is replaced by (II) and multiplying the resulting bacterium; (5) producing a recombinant microorganism by transferring (II) into a microorganism, replicating (II) in the microorganism and multiplying the resulting microorganism; (6) recombinant microorganism containing (II); (7) vector containing (II); (8) recombinant microorganism containing the vector; (9) producing a L-amino acid by culturing a coryneform bacterium mutant as above so that the L-amino acid is enriched in the culture broth or in the bacterial cells; (10) animal feed additive based on a culture broth, with a lysine content of 10-73 wt.%, a water content of up to 5 wt.% and a biomass content that is at least 0.1% of the biomass in the culture broth, where the biomass is derived from a bacterium as above; (11) animal feed additive based on a culture broth, with a tryptophan content of at least 5 wt.%, a water content of up to 5 wt.% and a biomass content that is at least 0.1% of the biomass in the culture broth, where the biomass is derived from a bacterium as above.
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机译：棒状细菌突变体包含编码2-甲基柠檬酸脱水酶多肽（I）的基因，该基因在272位具有L-脯氨酸以外的蛋白原性氨基酸。独立权利要求还包括：（1）通过处理棒状细菌产生的棒状细菌突变体用诱变剂分离和繁殖所得的突变体，从突变体制备核酸，使用引物通过PCR使用核酸产生核酸分子，所述引物包含来自SEQ ID NO：1和1563位之间的序列的至少15个连续核苷酸NO：3或SEQ ID NO：7（说明书中未给出）和引物，该引物包含来自SEQ ID NO：3或SEQ ID NO：7的位置3000与1567之间互补序列的至少15个连续核苷酸。酸分子，并确定编码的氨基酸序列，可选地将该序列与SEQ ID NO：6（说明书中未给出）进行比较，并鉴定突变体ng编码在272位上具有除L-脯氨酸以外的蛋白原性氨基酸的多肽的多核苷酸; （2）分离的编码（I）的多核苷酸（II）; （3）分离的多核苷酸，其编码与SEQ ID NO：2的19-57位（在说明书中未给出）相对应的氨基酸序列，在272位具有L-脯氨酸以外的蛋白原性氨基酸。 （4）通过将（II）转移到棒状细菌中，从而用（II）代替细菌的2-甲基柠檬酸脱水酶基因，并繁殖所得的细菌，从而产生重组棒状细菌; （5）通过将（II）转移到微生物中，在该微生物中复制（II）并繁殖所得的微生物来产生重组微生物; （6）含有（II）的重组微生物; （7）含有（II）的载体; （8）含有该载体的重组微生物; （9）通过培养如上所述的棒状细菌突变体产生L-氨基酸，从而使L-氨基酸富集在培养液或细菌细胞中; （10）基于培养液的动物饲料添加剂，其赖氨酸含量为10-73 wt。％，水含量最高为5 wt。％，且生物质含量至少为培养物中生物质的0.1％肉汤，其中生物质来自上述细菌; （11）基于培养液的动物饲料添加剂，其色氨酸含量至少为5 wt。％，水含量最多为5 wt。％，且生物质含量至少为培养物中生物质的0.1％肉汤，其中生物质来自上述细菌。
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