Candida tropicalis cells and use thereof

Abstract

The invention relates to genetically engineered Candida tropicalis cells, use thereof and a method of production of ω-hydroxycarboxylic acids and ω-hydroxycarboxylic acid esters.

1 . A mutant  Candida tropicalis  cell, which has, compared with a wild type  Candida tropicalis  cell, a reduced activity of at least one of enzyme that is encoded by an intron-free nucleic acid sequence selected from groups A) and B)
 A) Seq ID No. 1, Seq ID No. 3, Seq ID No. 5, Seq ID No. 7, Seq ID No. 9, Seq ID No. 11, Seq ID No. 13, Seq ID No. 15, Seq ID No. 17, Seq ID No. 19, Seq ID No. 21, Seq ID No. 23, Seq ID No. 25, Seq ID No. 27, Seq ID No. 29, Seq ID No. 31, Seq ID No. 33, Seq ID No. 35, Seq ID No. 37, Seq ID No. 39, Seq ID No. 41, Seq ID No. 43, Seq ID No. 45, Seq ID No. 47, Seq ID No. 49, Seq ID No. 51, Seq ID No. 53, Seq ID No. 55, Seq ID No. 57, Seq ID No. 59, Seq ID No. 61, Seq ID No. 63, Seq ID No. 65 and Seq ID No. 67   B) a sequence that is 80% identical to at least to one sequence selected from the group consisting of Seq ID No. 1, Seq ID No. 3, Seq ID No. 5, Seq ID No. 7, Seq ID No. 9, Seq ID No. 11, Seq ID No. 13, Seq ID No. 15, Seq ID No. 17, Seq ID No. 19, Seq ID No. 21, Seq ID No. 23, Seq ID No. 25, Seq ID No. 27, Seq ID No. 29, Seq ID No. 31, Seq ID No. 33, Seq ID No. 35, Seq ID No. 37, Seq ID No. 39, Seq ID No. 41, Seq ID No. 43, Seq ID No. 45, Seq ID No. 47, Seq ID No. 49, Seq ID No. 51, Seq ID No. 53, Seq ID No. 55, Seq ID No. 57, Seq ID No. 59, Seq ID No. 61, Seq ID No. 63, Seq ID No. 65 and Seq ID No. 67.   
     
     
         2 . The  Candida tropicalis  cell according to  claim 1 , wherein the decrease in enzymatic activity is achieved by modification of the nucleic acid sequence in the cell, wherein the modification is selected from the group consisting of insertion of foreign DNA into the nucleic acid sequence in the cell, deletion of at least parts of the nucleic acid sequence in the cell, a point mutation in the nucleic acid sequence in the cell, subjecting the nucleic acid sequence in the cell to RNA interference and exchanging a part of the nucleic acid sequence in the cell with foreign DNA. 
     
     
         3 . The  Candida tropicalis  cell according to  claim 2 , wherein the foreign DNA is a selection marker gene. 
     
     
         4 . The  Candida tropicalis  cell according to  claim 1 , wherein the cell is blocked at least partially in its β-oxidation. 
     
     
         5 . The  Candida tropicalis  cell according to  claim 1 , which is derived from a strain selected from the group consisting of  Candida tropicalis  H41,  Candida tropicalis  H41B,  Candida tropicalis  H51,  Candida tropicalis  H45,  Candida tropicalis  H43,  Candida tropicalis  H53,  Candida tropicalis  H534,  Candida tropicalis  534B,  Candida tropicalis  H435,  Candida tropicalis  ATCC20962 and  Candida tropicalis  HDC100. 
     
     
         6 . The  Candida tropicalis  cell according to  claim 5 , which is derived from  Candida tropicalis  ATCC20962 or  Candida tropicalis  HDC100. 
     
     
         7 . A method for producing ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid ester, the method comprising
 a) contacting the  Candida tropicalis  cell according to  claim 1  with a medium comprising a carboxylic acid or a carboxylic acid ester, 
 b) cultivating the cell under conditions to form the corresponding ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid esters from the carboxylic acid or the carboxylic acid ester and 
 c) optionally isolating the ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid esters that formed. 
 
     
     
         8 . A method of producing a  C. tropicalis  cell according to  claim 1 , the method comprising:
 I) preparing a  C. tropicalis  cell and   II) modifying at least one gene comprising one of the sequences selected from the nucleic acid sequence groups A) and B) stated in  claim 1  by insertion of foreign DNA into the gene, deletion at least of a part of the gene, a point mutation in the gene sequence, subjecting the gene to RNA interference and exchanging a part of the gene with foreign DNA.   
     
     
         9 . The method according to  claim 7 , wherein the ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid ester is a ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid ester with a chain length of the carboxylic acid from 6 to 24 carbon atoms and a chain length of the alcohol component of the ester from 1 to 4 carbon atoms 
     
     
         10 . The method according to  claim 7 , wherein the ω-hydroxycarboxylic acid or ω-hydroxycarboxylic acid ester is a 12-hydroxydodecanoic acid or 12-hydroxydodecanoic acid methyl ester. 
     
     
         11 . A method according to  claim 7 , wherein the  Candida tropicalis  cells are derived from a strain selected from the group consisting of  Candida tropicalis  H41,  Candida tropicalis  H41B,  Candida tropicalis  H51,  Candida tropicalis  H45,  Candida tropicalis  H43,  Candida tropicalis  H53,  Candida tropicalis  H534,  Candida tropicalis  534B,  Candida tropicalis  H435,  Candida tropicalis  ATCC20962 and  Candida tropicalis  HDC100 and wherein the cells are at least partially blocked in their β-oxidation. 
     
     
         12 . A method of manufacturing a polymer, the method comprising polymerizing the ω-hydroxycarboxylic acid or of the ω-hydroxycarboxylic acid ester obtained by the method according to  claim 7 .