Biocatalytic methods to convert cyclohexane oxidation process waste streams to useful products

Abstract

The invention relates to methods for enriching monomer content in a cycloalkane oxidation process mixed organic waste stream. In particular, the methods involve combining a biocatalyst with a mixed organic waste stream from a cycloalkane oxidation process, and enzymatically converting dimeric and/or oligomeric components of said waste stream into monomeric components. The methods may enrich the content of diacids, adipic acid, and/or other α,ω-difunctional C6 alkanes in the mixed organic waste stream. Additionally, the treated mixed organic waste streams may have improved burning efficiency.

1 .- 58 . (canceled) 
     
     
         59 . A recombinant host cell comprising at least one exogenous nucleic acid encoding a hydrolase classified under EC 3.1.1.-, wherein the hydrolase classified under EC 3.1.1.-, when contacted with a mixed organic waste stream from a cycloalkane oxidation process, hydrolyzes at least a portion of dimeric, oligomeric or cyclic esters of the mixed organic waste stream into aliphatic monomeric components. 
     
     
         60 . The recombinant host cell of  claim 59 , wherein the hydrolase classified under EC 3.1.1.- is selected from a lipase classified under EC 3.1.1.3, an esterase classified under EC 3.1.1.1, a cutinase classified under EC 3.1.1.74, a polyhydroxyalkanoate (PHA) depolymerase classified under EC 3.1.1.75 or EC 3.1.1.76, a lactone hydrolase classified under EC 3.1.1.25, and a gluconolactonase classified under EC 3.1.1.17. 
     
     
         61 . The recombinant host cell of  claim 59 , further comprising one or more enzymes from an endogenous or heterologous ω-oxidation pathway. 
     
     
         62 . The recombinant host cell of  claim 61 , wherein the one or more enzymes from the endogenous or heterologous ω-oxidation pathway is selected from:
 a P450 cytochrome oxidase, an ω-hydroxylase, ω-oxygenase enzyme or alkane-1-monooxygenase from the class EC 1.14.15.3; and 
 a fatty alcohol oxidase or an alcohol dehydrogenase from the class EC 1.1.1-. 
 
     
     
         63 . The recombinant host cell of  claim 59 , further comprising one or more enzymes from an endogenous or heterologous cyclohexanol and/or cyclohexane-1,2-diol degradation pathway. 
     
     
         64 . The recombinant host cell of  claim 63 , wherein the one or more enzymes from the endogenous or heterologous cyclohexanol degradation pathway is selected from:
 a cyclohenol dehydrogenase (ChnA) classified under EC 1.1.1.245, a cyclohexanone monooxygenase (ChnB) classified under EC 1.14.13.22, a 6-hexanolide hydrolase (ChnC) classified under EC 3.1.1.-, a 6-hydroxyhexanoate dehydrogenase (ChnD) classified under EC 1.1.1.258, and a 6-oxohexanoate dehydrogenase (ChnE) classified under EC 1.1.1.63.   
     
     
         65 . The recombinant host cell of  claim 63 , wherein the one or more enzymes from the endogenous or heterologous cyclohexane-1,2-diol degradation pathway is selected from:
 a cyclohexane-1,2-diol dehydrogenase classified under EC 1.1.1.174 and either a 2-hydroxycyclohexanone 2-monooxygenase classified under EC 1.14.13.66 or a cyclohexane-1,2-dione hydrolase classified under EC 3.7.1.11 or EC 3.7.1.10.   
     
     
         66 . The recombinant host cell of  claim 59 , further comprising a carboxylate reductase classified under EC 1.2.99.6, or an aldehyde dehydrogenase classified under EC 1.2.1.3 or EC 1.2.1.31, wherein the recombinant host cell converts 6-oxohexanoic acid to adipic acid. 
     
     
         67 . The recombinant host cell of  claim 59 , further comprising a 6-hydroxyhexanoate dehydrogenase classified under EC 1.1.1.258, and/or an alcohol/aldehyde dehydrogenase classified under EC 1.2.1.10, wherein the recombinant host cell converts 6-oxohexanoic acid to 1,6-hexanediol. 
     
     
         68 . The recombinant host cell of  claim 59 , further comprising an ω-aminotransferase classified under EC 2.6.1.191, wherein the recombinant host cell converts 6-oxohexanoic acid to 6-aminocaproic acid. 
     
     
         69 . The recombinant host cell of  claim 68 , further comprising an amidohydrolase classified under EC 3.5.2.-, wherein the recombinant host cell converts 6-aminocaproic acid to caprolactam. 
     
     
         70 . The recombinant host cell of  claim 68 , further comprising an aldehyde dehydrogenase classified under EC 1.2.1.3 or EC 1.2.1.31, and/or an aminotransferase classified under 2.6.1.191, wherein the recombinant host cell converts 6-aminocaproic acid to hexamethylenediamine. 
     
     
         71 . The recombinant host cell of  claim 59 , further comprising a laccase classified under EC 1.10.3.2. 
     
     
         72 . The recombinant host cell of  claim 59 , wherein the recombinant host cell has a deficiency in beta oxidation activity, adipate CoA ligase activity, CoA transferase activity, and/or acyl-CoA oxidase activity. 
     
     
         73 . The recombinant host cell of  claim 59 , wherein the recombinant host cell is a prokaryote. 
     
     
         74 . The recombinant host cell of  claim 73 , wherein said prokaryote is selected from  Escherichia, Clostridia, Corynebacteria, Cupriavidus, Pseudomonas, Delftia, Bacillus, Lactobacillus, Lactococcus, Rhodococcus, Marinobacter, Acinetobacter, Oleiphilus, Arthrobacter, Sphingomona, Burkholderia , and  Alcanivorax.    
     
     
         75 . The recombinant host cell of  claim 74 , wherein said prokaryote is selected from  Escherichia coli, Clostridium ljungdahlii, Clostridium autoethanogenum, Clostridium kluyveri, Corynebacterium glutamicum, Cupriavidus necator, Cupriavidus metallidurans, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas oleavorans, Delftia acidovorans, Bacillus subtillis, Lactobacillus delbrueckii, Lactococcus lactis, Rhodococcus erythropolis. Pseudomonas aeroginosa, Marinobacter hydrocarbonoclasticus, Acinetobacter venetianus, Oleiphilus messinensis, Arthrobacter viscosus, Sphingomona spaucimobilis, Burkholderia cepacia , and  Alcanivorax diesolei.    
     
     
         76 . The recombinant host cell of  claim 59 , wherein the recombinant host cell is a eukaryote. 
     
     
         77 . The recombinant host cell of  claim 76 , wherein said eukaryote is selected from  Aspergillus, Yarrowia, Issatchenkia, Debaryomyces, Arxula, Kluyveromyces, Candida, Rhodotorula, Rhizopus, Trichosporon, Trichoderma , and  Lipomyces.    
     
     
         78 . The recombinant host cell of  claim 77 , wherein said eukaryote is selected from  Aspergillus niger, Yarrowia lipolytica, Issathenkia orientalis, Debaryomyces hansenii, Arxula adenoinivorans, Kluyveromyces lactis, Candida albicans, Candida cloacae, Candida guillermondii, Candida intermedia, Candida maltosa, Candida parapsilosis, Candida zeylenoides , and  Trichoderma reesii.