AU5978200A - Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbial production of folic acid - Google Patents

Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbial production of folic acid Download PDF

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AU5978200A
AU5978200A AU59782/00A AU5978200A AU5978200A AU 5978200 A AU5978200 A AU 5978200A AU 59782/00 A AU59782/00 A AU 59782/00A AU 5978200 A AU5978200 A AU 5978200A AU 5978200 A AU5978200 A AU 5978200A
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polypeptide
folic acid
seq
amino acids
deletion
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Karin Herbster
Matthias Mack
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Sygnis Pharma AG
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Axaron Bioscience AG
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/34—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Corynebacterium (G)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1235—Diphosphotransferases (2.7.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C—CHEMISTRY; METALLURGY
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
    • C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system

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Description

0091/00003 Genes from Corynebacterium glutamicum for folic acid biosynthesis and their use for the microbial production of folic acid 5 The present invention is concerned with the process for producing folic acid by fermentation using a geneti cally manipulated organism. This invention consists of the nucleotide sequences of four genes (folE, folP, folB and folK) from Corynebacterium glutamicum for 10 folic acid biosynthesis and the use thereof for the microbial production of folic acid. These four genes form an operon and are transcribed in the following sequence: folE, folP, folB, folK. 15 Folic acid is essential for animal organisms. Its derivative tetrahydrofolate is a very versatile carrier of activated one-carbon units in cells of the animal organism. Folic acid consists of three groups: a sub stituted pteridine ring, p-aminobenzoate and glutamate. 20 Mammals are unable to synthesize a pteridine ring. They absorb folic acid from the diet and from microorganisms in their intestinal tract. Folic acid deficiency leads mainly to lesions in the mucous membranes. 25 The commercial importance of folic acid is in the animal feed and human food markets. Folic acid is employed mainly as a dietary supplement. Microorganisms can be employed for the fermentative 30 production of folic acid. They can be optimized in their efficiency of folic acid biosynthesis by genetic manipulation of the folic acid biosynthetic pathway. Genetic manipulation means in this connection that the number of copies and/or the rate of transcription of 35 the genes of the folic acid biosynthetic pathway is increased. As a consequence thereof, the proportion of gene product increases, and thus the intracellular enzymic activity does too. Increased enzymic activity leads to an increased rate of dietary (e.g. glucose) 0091/00003 - 2 conversion into folic acid and thus also to an increased product concentration. For genetic manipula tion, the nucleotide sequences of the genes of the folic acid biosynthetic pathway must be identified. 5 This invention is concerned with four novel gene sequences for folic acid biosynthesis from Corynebacterium glutamicum and with their use for the microbial production of folic acid. 10 One part of the invention comprises the folE gene product. SEQ ID NO. 2 describes a polypeptide sequence. The folE gene product encodes a polypeptide of 202 amino acids with a molecular weight of 22 029 Da. The present invention is also concerned with functional 15 derivatives of this polypeptide obtainable by replacing one or more amino acids, preferably up to 25% of the amino acids, most suitably up to 15% of the amino acids, in SEQ ID NO. 2 by deletion, insertion or sub stitution or by a combination of deletion, insertion 20 and substitution. The term functional derivative means that the enzymatic activity of the derivative is still of the same order of magnitude as that of the poly peptide having the sequence SEQ ID NO. 2. Another part of the invention comprises the folP gene 25 product. SEQ ID NO. 4 describes a polypeptide sequence. The folP gene product encodes a polypeptide of 285 amino acids with a molecular weight of 29 520 Da. The present invention is also concerned with functional derivatives of this polypeptide obtainable by replacing 30 one or more amino acids, preferably up to 40% of the amino acids, most suitably up to 25% of the amino acids, in SEQ ID NO. 4 by deletion, insertion or sub stitution or by a combination of deletion, insertion and substitution. The term functional derivative means 35 that the enzymatic activity of the derivative is still of the same order of magnitude as that of the poly peptide having the sequence SEQ ID NO. 4.
0091/00003 - 3 Another part of the invention comprises the folB gene product. SEQ ID NO. 6 describes a polypeptide sequence. The folB gene product encodes a polypeptide of 131 amino acids with a molecular weight of 14 020 Da. The 5 present invention is also concerned with functional derivatives of this polypeptide obtainable by replacing one or more amino acids, preferably up to 30% of the amino acids, most suitably up to 20% of the amino acids, in SEQ ID NO. 6 by deletion, insertion or sub 10 stitution or by a combination of deletion, insertion and substitution. The term functional derivative means that the enzymatic activity of the derivative is still of the same order of magnitude as that of the poly peptide having the sequence SEQ ID NO. 6. 15 Another part of the invention comprises the folK gene product. SEQ ID NO. 8 describes a polypeptide sequence. The folK gene product encodes a polypeptide of 160 amino acids with a molecular weight of 18 043 Da. The 20 present invention is also concerned with functional derivatives of this polypeptide obtainable by replacing one or more amino acids, preferably up to 40% of the amino acids, most suitably up to 30% of the amino acids, in SEQ ID NO. 8 by deletion, insertion or sub 25 stitution or by a combination of deletion, insertion and substitution. The term functional derivative means that the enzymatic activity of the derivative is still of the same order of magnitude as that of the poly peptide having the sequence SEQ ID NO. 8. 30 Another part of the invention comprises the polynucleo tide sequences which encode the polypeptides described above. The polynucleotide sequences can be generated starting from sequences isolated from Corynebacterium 35 glutamicum (i.e. SEQ ID NOS. 1, 3, 5 and 7) in which these sequences are modified by site-directed mutagenesis or a total chemical synthesis is carried out after back-translation of the corresponding 0091/00003 - 4 polypeptide using the genetic code. These polynucleotide sequences can preferably be employed for the transformation of host organisms, and 5 in this connection preferably of microorganisms, specifically in the form of gene constructs which comprise at least one copy of one of these polynucleo tides together with at least one regulatory sequence. Regulatory sequences comprise promoters, terminators, 10 enhancers and ribosome binding sites. Preferred host organisms for transformation with these gene constructs are Corynebacterium and Bacillus species. It is also possible to employ any eukaryotic 15 microorganism, preferably yeast strains of the genus Ashbya, Candida, Pichia, Saccharomyces and Hansenula. Another part of the invention comprises the process for producing folic acid by cultivating a host organism 20 which is transformed in the manner described above, and subsequently isolating the folic acid. The processes and the procedures for cultivating micro organisms and for isolating folic acid from a microbial 25 production are familiar to trained staff. The invention is described in more detail in the following examples, as is its use for the genetic manipulation of microorganisms, to increase the 30 efficiency of folic acid synthesis. Example 1 Construction of a genome library from Corynebacterium 35 glutamicum ATCC 13032 DNA from the genome of Corynebacterium glutamicum ATCC 13032 can be obtained by standard methods which 0091/00003 - 5 have already been described, for example by J. Altenbuchner and J. Cullum (1984, Mol. Gen. Genet. 195: 134-138). The genome library can be produced in accordance with standard protocols (e.g. Sambrook, J. 5 et al. (1989) Molecular cloning: a laboratory manual, Cold Spring Harbor Laboratory Press) with any cloning vector, e.g. pBluescript II KS- (Stratagene) or ZAP ExpressTM. (Stratagene) . It is moreover possible to use any fragment size, preferably Sau3AI fragments with a 10 length of 2-9 kb, which can be incorporated into cloning vectors with digested BamHI. Example 2 15 Analysis of the nucleic acid sequence of the genome library Individual E. coli clones can be selected from the genome library constructed in example 1. E. coli cells 20 are cultivated by standard methods in suitable media (e.g. LB supplemented with 100 mg/l ampicillin), and the plasmid DNA can then be isolated. Cloning of genome fragments from the DNA of Corynebacterium glutamicum into pBluescript II KS- (see example 1) allows the DNA 25 to be sequenced with the aid of the oligonucleotides 5'-AATTAACCCTCACTAAAGGG-3' and 5' -GTAATACGACTCACTATAGGGC-3'. Example 3 30 Computer analysis of the sequences of the isolated nucleic acids The nucleotide sequences can be connected together for 35 example with the aid of the BLASTX algorithm (Altschul et al. (1990) J. Mol. Biol. 215: 403-410). It is possible in this way to discover novel sequences and elucidate the function of these novel genes.
0091/00003 - 6 Example 4 Identification of an E. coli clone which comprises a 5 nucleotide sequence of the gene for GTP cyclo hydrolase I (EC 3.5.4.16) Analysis of the E. coli clones as described in example 2, which was followed by analysis, as described in example 3, of the sequences obtained thereby 10 revealed a sequence which is described by SEQ ID NO. 1. On use of the BLASTX algorithm (see example 3), this sequence revealed similarity with GTP cyclohydrolases I (FolE; EC 3.5.4.16) from various organisms. The greatest similarity was with the GTP cyclohydrolase I 15 (FolE) from Mycobacterium tuberculosis (NRDB 006273; 72% agreement at the amino acid level). Example 5 20 Identification of an E. coli clone which comprises a nucleotide sequence of the gene for dihydropteroate synthase (EC 2.5.1.15) Analysis of the E. coli clones as described in 25 example 2, which was followed by analysis, as described in example 3, of the sequences obtained thereby revealed a sequence which is described by SEQ ID NO. 3. On use of the BLASTX algorithm (see example 3), this sequence revealed similarity with dihydropteroate 30 synthases (FolP; EC 2.5.1.15) from various organisms. The greatest similarity was with the dihydropteroate synthase (FolP) from Mycobacterium tuberculosis (NRDB 006274; 53% agreement at the amino acid level).
0091/00003 - 7 Example 6 Identification of an E. coli clone which comprises a nucleotide sequence of the gene for dihydroneopterin 5 aldolase (EC 4.1.2.25) Analysis of the E. coli clones as described in example 2, which was followed by analysis, as described in example 3, of the sequences obtained thereby 10 revealed a sequence which is described by SEQ ID NO. 5. On use of the BLASTX algorithm (see example 3), this sequence revealed similarity with dihydroneopterin aldolases (FolB; EC 4.1.2.25) from various organisms. The greatest similarity was with the dihydroneopterin 15 aldolase (FolB) from Mycobacterium tuberculosis (NRDB 006275; 61% agreement at the amino acid level). Example 7 20 Identification of an E. coli clone which comprises a nucleotide sequence of the gene for 2-amino-4-hydroxy 6-hydroxymethyldihydropteridine pyrophosphokinase (EC 2.7.6.3) 25 Analysis of the E. coli clones as described in example 2, which was followed by analysis, as described in example 3, of the sequences obtained thereby revealed a sequence which is described by SEQ ID NO. 7. On use of the BLASTX algorithm (see example 3), this 30 sequence revealed similarity with 2-amino-4-hydroxy 6-hydroxymethyldihydropteridine pyrophosphokinases (FolK; EC 2.7.6.3) from various organisms. The greatest similarity was with the 2-amino-4-hydroxy-6-hydroxy methyldihydropteridine pyrophosphokinase (FolK) from 35 Mycobacterium leprae (EMBL AL023093; 43% agreement at the amino acid level).
0091/00003 - 8 Example 8 Use of the genes for GTP cyclohydrolase I, for dihydropteroate synthase, for dihydroneopterin aldolase 5 and for 2-amino-4-hydroxy-6-hydroxymethyldihydro pteridine pyrophosphokinase from Corynebacterium glutamicum for producing folic acid The genes for GTP cyclohydrolase I, for dihydropteroate 10 synthase, for dihydroneopterin aldolase and for 2-amino-4-hydroxy-6-hydroxymethyldihydropteridine pyrophosphokinase from Corynebacterium glutamicum can be introduced with the aid of suitable cloning and expression systems into Corynebacterium glutamicw or 15 into any other microorganism. Genetically manipulated microorganisms which differ from the wild-type organism in relation to the activity or the number of gene copies can be produced. These novel genetically manipulated strains can be employed for producing folic 20 acid. Sequence list (I) General information 25 (1) Applicant: (A) Name: BASF-LYNX Bioscience AG (B) Street: Im Neuenheimer Feld 515 30 (C) City: Heidelberg (D) Country: Germany (E) Postal code: 69120 (F) Telephone: 06221/4546 (G) Fax: 06221/454770 35 (2) Title: Genes from Corynebacterium glutamicum for folic acid biosynthesis and their use for the microbial production of folic acid 0091/00003 - 9 (3) Number of sequences. 8 SEQ ID NO. 1: DNA (folE) 5 AkTGAAGGAGACAACCGTGGAT7AACCACGCTGCAGTTCCCGAGTT4CGATGAGGAGCGCGCAACAGC CAGCTCGAGTGGCTAGGGCGTACAAGGACTTTCGCGGGTCTGCATGAGGATCCCACCACTGTG CATGTGT1GAGCACC ACTTGGTGCCGTTCTTTGGCGTGGCGCACAT~TGGTTACATTCCGGGTAAGT GTTCAGGAGCGC TACCCATrC(PTCCCGC-AACTAGCAICG GGCCGTGGTGTGAAGCTGAGCACCCTCAGGCCATGCCCC-TCCTAAGCCTGGTGCTC TCCCTGA7TCGGGGGCACTA.k SEQ ID NO. 2: amino acid (FolE) LEKTP SEGHEELVLREI P £YSMCEIiKLNVPFFGVAXIGYI PGKSGKTGLSI(IARLADFAKPQ VQERL'VSQ AA-EJIA~VVZAHCAXGRPATTARGKASA~ 10 SIG SEQ ID NO. 3: DNA (foiP) AT~ACTTCCTGCACCGAGTTTTAGIATGCAGCCG GGTCTTCGCGGCAGAATA-TGTAGGTI;GAGCAGA TGTGTCGCCCCTA'GTTGCGGbGCACGCTGGAGGG GTGCCTCTGAGGCGGTTCGTATAGGTCCAGCGA CCAATCG AAC GCGCTCTGGAGTCCGGGCCGCTTC TGTACAGCCGCGTGCG'rCGGTTTCGCTGGAGGA ATCCTTTTAGATGGXCT'~AT:GGT;CCCiTAGAAC CGGTGGAGACGTTAGCCGTCACGCAGTGTTATGTCTTCGCCCGCGCCACCGCTG CTGGGCG.ACAACMGTGTCAGTGGTTCAACCTA4A A~CWG~G~GACATCCGGTA-TCGACT4CCkCT-TCGO ATCCCGGAGCGATTCCTGQC^TGGCCGCGCAAAACCGTGGCCTAIGTGTCACCCCCATTGATG CCACACACCGGTACCGGCTCCTTGACTGGTTCCT CACGATGPC CCGiKCAGAGTTGTTGCCTGGCAGGAG.CC C CATGGCTGA 0091/00003 - 10 SEQ ID NO. 4: amino acid (FoiP) MNVSSLt'r PGRCL VM'G rTV'EDFSDCKYZDVI)QA kAYELVAACADXI DVGGESTRPGAJR VDASVERDRVVFKAL~rAGiTTFSVAQkAAGAGVSINDVSGGLADPEb'MAEQ I VLO TQGAG~)GDVDHVDDLAT.GANILP~,~KR. N\WRLLQALP EFI SGPF PIL ,SYRLGRK)GIVPDDATATVAMAG aDVPVSRDVVAIWRSGGTHVHG 5 SEQ ID NO. 5: DNA (FoiB) ATGGCTGATCGTATTGkACTTAAAGGCCTTGAATGCTTCGGACACCACGGTGTGTTCGACTTTGA AAAAGAGCAAGGCCAGCCCTTCATTGTGGATGTCACCTGCTGGATGGATTTCGATGCCGCAGGTG CCCCAGCTTC CCGAATCGCCTGCTTCTGTAACT GAGGCCCATCCAGGGATTTGTCGAGACGGTGGCCAOAATCTGCGATGCTTGATGGCTAA ATGATGCGCTTCATGCGGTGGAMGTAACCATCCATAGCCCAAAGCACCGATCCCACGTACTT TTGCTG-ACGTCGCC-GTGGTTGCCCGACGTTLCCACGAAATCCATG;GCTGCTGGAAGG;AGCAACGCC TA.A SEQ ID NO. 6: amino acid (FoiB) 10 DALRIELKGLECFGIEHGVFDFEKEQGQ PF IVDVTPCWCFDAAGASDDILSD)TVDYGALALLV7A.EIV EGPSRDL IETVATESADAVMA (FDALHAVEVTIHKPK~AP IPRVAVARSRKSMAAC.SNA SEQ ID NO. 7: DNA (folK) .ATGCATGCAGTTTTCTCCATCGGTTCCAACATGGATGATCGCTACGCGCTJGCTCAACACAGTGAT CGAGGA6ATCAAGAGAGTCGTGGCGCAGTCTGC-krCTACTCAACCCCACCGTGGGGCATTG AGGATCAGGATGAAkTTCCVrCAACGCAGTGYCTCGT2-GTTGAGGTTCrAAGMAACCCCCATCCGAGTTG CTGCGCCGTQcGCCAA7AACTCGAAGAAGCCGCCGAGCGGG1'CCGCCTCCGCAAATGGGGGCCACG CACCCTCGA2'CTGGATATCGTG-CAGATCATTAAAGATGGGGAAGAGATCCTTITCTGAGGATCCCG AACTGACCTTGCCACACCCTTCGGCTTGGCACGCGTGCCETCGTGTTGATCCCTTGG-TTGGAAGCA GAACCTGATGCCGTCCTGCACC-GCACGACCATTCAC-AACATGTGGATAATCTTGATCCCALCAGA 15 CATTGAAGGTGTCACCA.AGATTTAA SEQ ID NO. 8: amino acid (folK) MVLSICSNIDRYALNTVIEFFKDE IVAQSM.YSTI'PWG-IEDQD)EFLNAVLVVEVEETPIEL LRRGQKLEEAAERVRVRWGPRTLD.?DIVIQIIDGEEILSEI)PELTLPHFWAWQRAFVLIPWLEA E PDPVLHG'TA~h'vNL0PTD IEGVTKI

Claims (8)

1. A polypeptide having GTP cyclohydrolase I activity and selected from the following group: 5 (a) a polypeptide having the amino acid sequence which is described in SEQ ID NO. 2 (b) a polypeptide which is modified by comparison 10 with that in (a) by deletion, insertion or substitution of one or more amino acids.
2. A polypeptide having dihydropteroate synthase activity and selected from the following group: 15 (a) a polypeptide having the amino acid sequence which is described in SEQ ID NO. 4; (b) a polypeptide which is modified by comparison 20 with that in (a) by deletion, insertion or substitution of one or more amino acids.
3. A polypeptide having dihydroneopterin aldolase activity and selected from the following group: 25 (a) a polypeptide having the amino acid sequence which is described in SEQ ID NO. 6 (b) a polypeptide which is modified by comparison 30 with that in (a) by deletion, insertion or substitution of one or more amino acids.
4. A polypeptide having 2-amino-4-hydroxy-6-hydroxy methyldihydropteridine pyrophosphokinase activity 35 and selected from the following group: (a) a polypeptide having the amino acid sequence which is described in SEQ ID NO. 8 0091/00003 - 12 (b) a polypeptide which is modified by comparison with that in (a) by deletion, insertion or substitution of one or more amino acids. 5
5. A polynucleotide which encodes a polypeptide corresponding to claim 1, 2, 3 or 4.
6. A gene construct having at least one copy of a 10 polynucleotide corresponding to claim 5, together with at least one regulatory sequence.
7. A host organism which is transformed with a gene construct corresponding to claim 6. 15
8. A process for producing folic acid by cultivating a host organism corresponding to claim 7 with subsequent isolation of the folic acid.
AU59782/00A 1999-06-25 2000-06-23 Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbial production of folic acid Abandoned AU5978200A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19929363 1999-06-25
DE19929363A DE19929363A1 (en) 1999-06-25 1999-06-25 New Corynebacterium glutamicum GTP cyclohydrolase I, dihydropteroate synthase, dihydroneopterin aldolase and 2-amino-4-hydroxy-6-hydroxymethyl-dihydropteridine pyrophosphokinase polypeptides
PCT/EP2000/005864 WO2001000845A1 (en) 1999-06-25 2000-06-23 Genes from corynebacterium glutamicum for the biosynthesis of folic acid and their use for the microbial production of folic acid

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US6958228B2 (en) 2000-08-02 2005-10-25 Degussa Ag Nucleotide sequence which code for the metH gene
US6942996B2 (en) 2000-08-02 2005-09-13 Degussa Ag Isolated polynucleotide from Corynebacterium encoding a homocysteine methyltransferase
DE10039044A1 (en) 2000-08-10 2002-02-21 Degussa Novel polynucleotide from Coryneform bacteria coding for lysR1 gene, useful as hybridization probe for detecting DNA coding for transcription regulator lysR1
DE10039049A1 (en) 2000-08-10 2002-02-21 Degussa Novel polynucleotide from Coryneform bacteria coding for lysR3 gene, useful as a probe for detecting DNA to isolate nucleic acids coding for transcription regulator lysR3 or for producing L-amino acids, e.g., L-lysine and L-valine
DE10039043A1 (en) 2000-08-10 2002-02-21 Degussa New nucleotide sequences coding for the luxR gene
AU2001291658A1 (en) 2000-08-26 2002-03-13 Degussa A.G. Nucleotide sequences which code for the ccpa2 gene
US6815196B2 (en) 2000-09-02 2004-11-09 Degussa Ag Nucleotide sequences encoding o-succinylhomoserine sulfhydrylase
US6812016B2 (en) 2000-09-02 2004-11-02 Degussa Ag Nucleotide sequences which code for the metY gene
AU2001293723A1 (en) 2000-09-09 2002-03-22 Degussa A.G. Nucleotide sequences coding for the dep33 gene
US6759224B2 (en) 2000-09-09 2004-07-06 Degussa Ag Nucleotide sequences which code for the sahH gene
DE10045496A1 (en) 2000-09-14 2002-03-28 Degussa New nucleotide sequences coding for the ptsi gene
DE10055869A1 (en) 2000-11-10 2002-05-29 Degussa A polynucleotide encoding the nadA gene useful for the preparation of nicotinic acid or its derivatives, as probes for discovering RNA, cDNA and DNA to isolate polynucleotides or genes which code for quinolinate synthetase A
DE10055870A1 (en) 2000-11-10 2002-05-29 Degussa New nucleotide sequences coding for the nadC gene
EP1262541A1 (en) * 2001-05-28 2002-12-04 Stichting Top-Instituut Voedselwetenschappen Production of bioavailable folic acid
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