SYNERGISTIC BACTERIAL AND YEAST COMBINATIONS
The present disclosure concerns a symbiotic combination of host cells engineered to produce a first metabolic product, for example a carbohydrate, and to convert the second metabolic product into a second metabolic product, for example an alcohol.
1 . A combination of a first microbial host cell having a first metabolic pathway comprising one or more first enzymes for producing a first metabolic product and a second microbial host cell having a second metabolic pathway comprising one or more second enzymes for converting at least in part the first metabolic product into a second metabolic product, wherein:
at least one of the first microbial host cell or the second microbial host cell is recombinant;
at least one of the first microbial host cell or the second microbial host cell is a bacterial host cell;
at least one of the first microbial host cell or the second microbial host cell is a yeast host cell;
when the first microbial host cell is a recombinant first microbial host cell, the recombinant first microbial host cell has increased activity in the first metabolic pathway, when compared to a corresponding native first microbial host cell, for producing the first metabolic product; and
when the second microbial host cell is a recombinant second microbial host cell, the recombinant second microbial host cell has increased activity in the second metabolic pathway, when compared to a corresponding native second microbial host cell, for converting at least in part the first metabolic product into the second metabolic product.
2 . The combination of claim 1 , wherein the first microbial host cell is a bacterial host cell and the second microbial cell is a yeast host cell.
3 . The combination of claim 2 , wherein at least one of the one or more first enzymes are native enzymes and/or at least one of the one or more second enzymes are heterologous enzymes.
4 . (canceled)
5 . The combination of claim 2 , wherein the first metabolic product is an organic acid or an esther thereof and/or the second metabolic product is ethanol and wherein:
the one or more first enzymes comprises a citrate lyase, and/or
the one or more second enzymes comprise one or more of:
one or more heterologous polypeptides having acetaldehyde dehydrogenase activity, and/or one or more heterologous polypeptides having acetyl-coA synthetase activity.
6 .- 10 . (canceled)
11 . The combination of claim 5 , wherein (i) the yeast host cell is the recombinant yeast host cell and (ii) the heterologous polypeptide having acetaldehyde dehydrogenase activity is an acetylating dehydrogenase (AADH) or a bifunctional acetaldehyde/alcohol dehydrogenase (ADHE), the one or more second enzymes comprising an heterologous polypeptide having NADP + -dependent alcohol dehydrogenase activity and/or an heterologous polypeptide having acetyl-coA synthetase activity.
12 .- 13 . (canceled)
14 . The combination of claim 1 , wherein the first microbial host cell is a yeast host cell and the second microbial host cell is a bacterial host cell.
15 . The combination of claim 14 , wherein at least one of the one or more first enzymes is awe heterologous enzymes and/or at least one of the one or more second enzymes is a heterologous enzyme.
16 . (canceled)
17 . The combination of claim 14 , wherein the first metabolic product is a carbohydrate and/or the second metabolic product is ethanol.
18 . (canceled)
19 . The combination of claim 17 , wherein (i) the carbohydrate is trehalose and (ii) the one or more first enzymes comprises trehalose-6-phosphate synthase and/or trehalose-6-phosphate phosphatase.
20 .- 23 . (canceled)
24 . The combination of claim 17 , wherein (i) the carbohydrate is mannitol, and (ii) the one or more first enzymes comprises mannitol-1-phosphate 5-dehydrogenase, the one or more second enzymes comprise a product of at least one gene from a mannitol utilization operon, and/or the one or more second enzymes comprises a mannitol transporter.
25 .- 30 . (canceled)
31 . The combination of claim 17 , wherein (i) the carbohydrate is sorbitol and (ii) the one or more first enzymes comprises sorbitol-6-phosphate dehydrogenase, and/or the one or more second enzymes comprises a product of at least one gene from a sorbitol utilization operon.
32 .- 35 . (canceled)
36 . The combination of claim 17 or 18 , wherein (i) the carbohydrate is glycerol, (ii) the one or more second enzymes comprise at least one of a glycerol dehydrogenase, a dihydroxyacetone kinase, a glycerol kinase, a glycerol-3-phosphate dehydrogenase, and/or a glycerol facilitator.
37 .- 41 . (canceled)
42 . The combination of claim 36 , wherein the yeast host cell has increased activity, when compared to the corresponding native yeast host cell, in an NADP + -dependent aldehyde dehydrogenase and/or in a phosphoketolase.
43 .- 44 . (canceled)
45 . The combination of claim 1 , wherein the yeast host cell is from Saccharomyces sp. or from Saccharomyces cerevisiae.
46 . (canceled)
47 . The combination of claim 1 , wherein the bacterial host cell further comprises a third metabolic pathway comprising one or more third enzymes for producing a third metabolic product.
48 . (canceled)
49 . The combination of claim 47 , wherein the third metabolic product is ethanol and the one or more third enzymes for producing the third metabolic product comprises a pyruvate decarboxylase and/or an alcohol dehydrogenase; and/or wherein the bacterial host cell has a decreased lactate dehydrogenase activity when compared to the corresponding native bacterial host cell.
50 .- 59 . (canceled)
60 . The combination of claim 1 , wherein the bacterial host cell is a lactic acid bacteria.
61 . The combination of claim 60 , wherein the bacterial host cell is from Lactobacillus sp. or from Lactobacillus paracasei.
62 .- 65 . (canceled)
66 . A process for converting a biomass into a fermentation product, the process comprises contacting the biomass with the combination of claim 1 under condition to allow the conversion of at least a part of the biomass into the fermentation product.
67 .- 71 . (canceled)
72 . A commercial package comprising:
(i) a combination of a first microbial host cell having a first metabolic pathway comprising one or more first enzymes for producing a first metabolic product and a second microbial host cell having a second metabolic pathway comprising one or more second enzymes for converting at least in part the first metabolic product into a second metabolic product, wherein:
at least one of the first microbial host cell or the second microbial host cell is recombinant;
at least one of the first microbial host cell or the second microbial host cell is a bacterial host cell;
at least one of the first microbial host cell or the second microbial host cell is a yeast host cell;
when the first microbial host cell is a recombinant first microbial host cell, the recombinant first microbial host cell has increased activity in the first metabolic pathway, when compared to a corresponding native first microbial host cell, for producing the first metabolic product; and
when the second microbial host cell is a recombinant second microbial host cell, the recombinant second microbial host cell has increased activity in the second metabolic pathway, when compared to a corresponding native second microbial host cell, for converting at least in part the first metabolic product into the second metabolic product; and
(ii) instructions to perform a process for converting a biomass into a fermentation product, the process comprises contacting the biomass with the combination of (i) under condition to allow conversion of at least a part of the biomass into the fermentation product.
73 .- 75 . (canceled)