Isobutanol production using yeasts with modified transporter expression
View Patent ↗The present invention relates to recombinant microorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise one or more modifications resulting in the reduction in the expression and/or activity of an endogenous transporter protein. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.
1. A recombinant microorganism comprising enzymes of an isobutanol producing metabolic pathway, wherein said recombinant microorganism is a yeast microorganism and comprises at least one exogenous gene encoding a polypeptide that catalyzes a step in the conversion of pyruvate to isobutanol, wherein said recombinant microorganism is engineered to disrupt, partially delete, or completely delete a gene encoding OPT1, and wherein said recombinant microorganism produces more isobutanol than a corresponding yeast microorganism that has not been engineered to disrupt, partially delete, or completely delete a gene encoding OPT1.
2. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is engineered to disrupt, partially delete, or completely delete a gene encoding pyruvate decarboxylase.
3. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is engineered to disrupt, partially delete, or completely delete a gene encoding glycerol-3-phosphate dehydrogenase.
4. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is of the Saccharomyces clade.
5. The recombinant microorganism of claim 4 , wherein said recombinant microorganism is a Saccharomyces sensu stricto microorganism.
6. The recombinant microorganism of claim 5 , wherein said Saccharomyces sensu stricto microorganism is selected from the group consisting of S. cerevisiae, S. kudriavzevii, S. mikatae, S. bayanus, S. uvarum, S. carocanis and hybrids thereof.
7. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is a Crabtree-negative yeast microorganism.
8. The recombinant microorganism of claim 7 , wherein said Crabtree-negative yeast microorganism is classified into a genus selected from a group consisting of Saccharomyces, Kluyveromyces, Pichia, Hansenula, Issatchenkia and Candida.
9. The recombinant microorganism of claim 8 , wherein said Crabtree-negative yeast microorganism is selected from the group consisting of Saccharomyces kluyveri, Kluyveromyces lactis, Kluyveromyces marxianus, Pichia anomala, Pichia stipitis, Pichia kudriavzevii, Issatchenkia orientalis, Hansenula anomala, Candida utilis and Kluyveromyces waltii.
10. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is a Crabtree-positive yeast microorganism.
11. The recombinant microorganism of claim 10 , wherein said Crabtree-positive yeast microorganism is classified into a genus selected from a group consisting of Saccharomyces, Kluyveromyces, Zygosaccharomyces, Debaryomyces, Pichia, Candida , and Schizosaccharomyces.
12. The recombinant microorganism of claim 11 , wherein said Crabtree-positive yeast microorganism is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces uvarum, Saccharomyces bayanus, Saccharomyces paradoxus, Saccharomyces castelli, Kluyveromyces thermotolerans, Candida glabrata, Zygosaccharomyces bailli, Zygosaccharomyces rouxii, Debaryomyces hansenii, Pichia pastorius, Schizosaccharomyces pombe , and Saccharomyces uvarum.
13. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is a post-WGD (whole genome duplication) yeast microorganism.
14. The recombinant microorganism of claim 13 , wherein said post-WGD yeast microorganism is classified into a genus selected from a group consisting of Saccharomyces or Candida.
15. The recombinant microorganism of claim 14 , wherein said post-WGD yeast microorganism is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces uvarum, Saccharomyces bayanus, Saccharomyces paradoxus, Saccharomyces castelli , and Candida glabrata.
16. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is a pre-WGD (whole genome duplication) yeast microorganism.
17. The recombinant microorganism of claim 16 , wherein said pre-WGD yeast microorganism is classified into a genus selected from a group consisting of Saccharomyces, Kluyveromyces, Candida, Pichia, Debaryomyces, Hansenula, Issatchenkia, Pachysolen, Yarrowia and Schizosaccharomyces.
18. The recombinant microorganism of claim 17 , wherein said pre-WGD yeast microorganism is selected from the group consisting of Saccharomyces kluyveri, Kluyveromyces thermotolerans, Kluyveromyces marxianus, Kluyveromyces waltii, Kluyveromyces lactis, Candida tropicalis, Pichia pastoris, Pichia anomala, Pichia stipitis, Pichia kudriavzevii, Issatchenkia orientalis, Debaryomyces hansenii, Hansenula anomala, Pachysolen tannophilis, Yarrowia lipolytica , and Schizosaccharomyces pombe.
19. A method of producing isobutanol, comprising:
(a) providing a recombinant microorganism of claim 1 ;
(b) cultivating the recombinant microorganism in a culture medium containing a feedstock providing the carbon source, until a recoverable quantity of isobutanol is produced.