US 2007/0190605 A1 · GENE PRODUCTS OF BACILLUS LICHENIFORMIS WHICH FORM ODOROUS SUBSTANCES AND IMPROVED BIOTECHNOLOGICAL PRODUCTION METHODS BASED THEREON

Abstract

The present invention relates to 25 hitherto undescribed genes of B. licheniformis and gene products derived therefrom and all sufficiently homologous nucleic acids and proteins thereof. They occur in five different metabolic pathways for the formation of odorous substances. The metabolic pathways in question are for the synthesis of: 1) isovalerian acid (as part of the catabolism of leucine), 2) 2-methylbutyric acid and/or isobutyric acid (as part of the catabolism of valine and/or isoleucine), 3) butanol and/or butyric acid (as part of the metabolism of butyric acid), 4) propyl acid (as part of the metabolism of propionate) and/or 5) cadaverine and/or putrescine (as parts of the catabolism of lysine and/or arginine). The identification of these genes allows biotechnological production methods to be developed that are improved to the extent that, to assist these nucleic acids, the formation of the odorous substances synthesized via these metabolic pathways can be reduced by deactivating the corresponding genes in the micro-organism used for the biotechnological production. In addition, these gene products are thus available for preparing reactions or for methods according to their respective biochemical properties.

A nucleic acid selected from the group consisting of:

(a) nucleic acids coding for a gene product (putative branched-chain amino acid aminotransferase;
E.C. 2.6.1.42) involved in the synthesis of 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 67% identity to the nucleotide sequence indicated in SEQ ID NO. 1;
(b) nucleic acids coding for a gene product (putative branched-chain amino acid aminotransferase;
E.C. 2.6.1.42) involved in the synthesis of 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 78% identity to the nucleotide sequence indicated in SEQ ID NO. 3;
(c) nucleic acids speA coding for a gene product (lysine and/or arginine decarboxylase;
E.C. 4.1.1.18 or 4.1.1.19) involved in the synthesis of cadaverine and/or putrescine and having a nucleotide sequence which shows at least 78% identity to the nucleotide sequence indicated in SEQ ID No. 5;
(d) nucleic acids yugJ coding for a gene product (NADH-dependent butanol dehydrogenase A;
E.C. 1.1.1.-) involved in the synthesis of butanol and/or butyric acid and having a nucleotide sequence which shows at least 81% identity to the nucleotide sequence indicated in SEQ ID NO. 7;
(e) nucleic acids coding for a gene product (acyl-CoA dehydrogenase;
E.C. 1.3.99.-) involved in the synthesis of isovaleric acid, 2-methylbutyric acid, isobutyric acid, butanol and/or butyric acid and having a nucleotide sequence which shows at least 75% identity to the nucleotide sequence indicated in SEQ ID NO. 9;
(f) nucleic acids coding for a gene product (acyl-CoA dehydrogenase;
E.C. 1.3.99.-) involved in the synthesis of isovaleric acid, 2-methylbutyric acid, isobutyric acid, butanol and/or butyric acid and having a nucleotide sequence which shows at least 64% identity to the nucleotide sequence indicated in SEQ ID NO. 11;
(g) nucleic acids coding for a gene product (3-hydroxybutyryl-CoA dehydrogenase;
E.C. 1.1.1.157) involved in the synthesis of butanol and/or butyric acid and having a nucleotide sequence which shows at least 67% identity to the nucleotide sequence indicated in SEQ ID NO. 13;
(h) nucleic acids coding for a gene product (putative enoyl-Coa hydratase protein;
E.C. 4.2.1.17) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 65% identity to the nucleotide sequence indicated in SEQ ID NO. 15;
(i) nucleic acids coding for a gene product (probable enoyl-(3-hydroxyisobutyryl)-coenzyme A hydrolase protein) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 66% identity to the nucleotide sequence indicated in SEQ ID NO. 17;
(j) nucleic acids echA8 coding for a gene product (probable enoyl-CoA hydratase;
E.C. 4.2.1.17) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 48% identity to the nucleotide sequence indicated in SEQ ID NO. 19;
(k) nucleic acids coding for a gene product (acyl-CoA dehydrogenase;
E.C. 1.3.99.-) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 54% identity to the nucleotide sequence indicated in SEQ ID NO. 21;
(l) nucleic acids acsA coding for a gene product (acetyl-coenzyme A synthetase;
E.C. 6.2.1.1) involved in the synthesis of propionic acid and having a nucleotide sequence which shows at least 67% identity to the nucleotide sequence indicated in SEQ ID NO. 23;
(m) nucleic acids yngF coding for a gene product (3-hydroxybutyryl-CoA dehydratase;
E.C. 4.2.1.55) involved in the synthesis of butanol and/or butyric acid and having a nucleotide sequence which shows at least 68% identity to the nucleotide sequence indicated in SEQ ID NO. 25;
(n) nucleic acids yusJ coding for a gene product (acyl-CoA dehydrogenase;
E.C. 1.3.99.-) involved in the synthesis of isovaleric acid, 2-methylbutyric acid, isobutyric acid, butanol and/or butyric acid and having a nucleotide sequence which shows at least 77% identity to the nucleotide sequence indicated in SEQ ID NO. 27;
(o) nucleic acids ykwC coding for a gene product (hypothetical oxidoreductase;
E.C. 1.1.-.-) involved in the synthesis of 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 77% identity to the nucleotide sequence indicated in SEQ ID NO. 29;
(p) nucleic acids coding for a gene product (probable phosphate butyryltransferase;
E.C. 2.3.1.19) involved in the synthesis of butanol and/or butyric acid and having a nucleotide sequence which shows at least 51% identity to the nucleotide sequence indicated in SEQ ID NO. 31;
(q) nucleic acids coding for a gene product (probable butyrate kinase;
E.C. 2.7.2.7) involved in the synthesis of butanol and/or butyric acid and having a nucleotide sequence which shows at least 77% identity to the nucleotide sequence indicated in SEQ ID NO. 33;
(r) nucleic acids acsA coding for a gene product (acetyl-coenzyme A synthetase;
E.C. 6.2.1.1) involved in the synthesis of propionic acid and having a nucleotide sequence which shows at least 79% identity to the nucleotide sequence indicated in SEQ ID NO. 35;
(s) nucleic acids ytcl coding for a gene product (acetate-CoA ligase;
E.C. 6.2.1.1) involved in the synthesis of propionic acid and having a nucleotide sequence which shows at least 74% identity to the nucleotide sequence indicated in SEQ ID NO. 37;
(t) nucleic acids speA coding for a gene product (lysine and/or arginine decarboxylase;
E.C. 4.1.1.18 or E.C. 4.1.1.19) involved in the synthesis of cadaverine and/or putrescine and having a nucleotide sequence which shows at least 68% identity to the nucleotide sequence indicated in SEQ ID NO. 39;
(u) nucleic acids ysiB coding for a gene product (probable Enoyl-CoA hydratase;
E.C. 4.2.1.17) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 75% identity to the nucleotide sequence indicated in SEQ ID NO. 41;
(v) nucleic acids coding for a gene product (similar to 3-hydroxyacyl-CoA dehydrogenase;
E.C. 1.1.1.35) involved in the synthesis of 2-methylbutyric acid and having a nucleotide sequence which shows at least 76% identity to the nucleotide sequence indicated in SEQ ID NO. 43;
(w) nucleic acids coding for a gene product (2-oxoglutarate dehydrogenase E1 component;
E.C. 1.2.4.2) involved in the synthesis of isovaleric acid, 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 80% identity to the nucleotide sequence indicated in SEQ ID NO. 45;
(x) nucleic acids yhfL coding for a gene product (probable acid-CoA ligase;
E.C. 6.2.1.-) involved in the synthesis of propionic acid and having a nucleotide sequence which shows at least 67% identity to the nucleotide sequence indicated in SEQ ID NO. 47;
and
(y) nucleic acids ywhG coding for a gene product (agmatinase;
E.C. 3.5.1.11) involved in the synthesis of cadaverine and/or putrescine and having a nucleotide sequence which shows at least 85% identity to the nucleotide sequence indicated in SEQ ID NO. 49.

47 more elements

Gene products of bacillus licheniformis which form odorous substances and improved biotechnological production methods based thereon

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Gene products of bacillus licheniformis which form odorous substances and improved biotechnological production methods based thereon

Life of this patent

Abstract

Claims30