Biorefinery system, methods and compositions thereof

Abstract

The present disclosure relates to bioengineering approaches for producing biofuel and, in particular, to the use of a C 1 metabolizing microorganism reactor system for converting C 1 substrates, such as methane or methanol, into biomass and subsequently into biofuels, bioplastics, or the like.

What is claimed is: 
     
         1 . A C1 metabolizing non-photosynthetic microorganism biomass, wherein the δ 13 C of the biomass is less than about −30‰. 
     
     
         2 . The biomass according to  claim 1 , wherein the δ 13 C of the biomass ranges from about −40‰ to about −60‰. 
     
     
         3 . The biomass according to  claim 1 , wherein the C 1  metabolizing non-photosynthetic microorganism is a bacteria or a yeast. 
     
     
         4 . The biomass according to  claim 3 , wherein the C 1  metabolizing bacteria is a methanotroph or methylotroph. 
     
     
         5 . The biomass according to  claim 4 , wherein the bacteria is a  Methylomonas  sp. 16a (ATCC PTA 2402),  Methylosinus trichosporium  OB3b (NRRL B-11,196),  Methylosinus sporium  (NRRL B-11,197),  Methylocystis parvus  (NRRL B-11,198),  Methylomonas methanica  (NRRL B-11,199),  Methylomonas albus  (NRRL B-11,200),  Methylobacter capsulatus  Y (NRRL B-11,201),  Methylococcus capsulatus  Bath (NCIMB 11132),  Methylobacterium organophilum  (ATCC 27,886),  Methylomonas  sp. AJ-3670 (FERM P-2400),  Methylomicrobium alcaliphilum, Methylocella silvestris, Methylacidiphilum infernorum, Methylibium petroleiphilum, Methylobacterium extorquens, Methylobacterium radiotolerans, Methylobacterium populi, Methylobacterium chloromethanicum, Methylobacterium nodulans , or any combination thereof. 
     
     
         6 . The biomass according to  claim 3 , wherein the C 1  metabolizing bacteria is a syngas metabolizing bacteria. 
     
     
         7 . The biomass according to  claim 6 , wherein the syngas metabolizing bacteria is  Clostridium autoethanogenum, Clostridium ljungdahli, Clostridium ragsdalei, Clostridium carboxydivorans, Butyribacterium methylotrophicum, Clostridium woodii, Clostridium neopropanologen , or a combination thereof. 
     
     
         8 . The biomass according to  claim 1 , wherein the C 1  metabolizing non-photosynthetic microorganism is a recombinant C 1  metabolizing non-photosynthetic microorganism comprising a heterologous polynucleotide encoding a fatty acid producing enzyme, a formaldehyde assimilation enzyme, or any combination thereof. 
     
     
         9 . The biomass according to  claim 8 , wherein the recombinant C 1  metabolizing non-photosynthetic microorganism accumulates an increased level of fatty acids or overexpresses fatty acids as compared to a parent or reference C 1  metabolizing non-photosynthetic microorganism. 
     
     
         10 . The biomass according to  claim 8 , wherein the heterologous polynucleotide encodes a thioesterase, a malonyl CoA-acyl carrier protein transacylase, an acetyl-CoA carboxylase, or any combination thereof. 
     
     
         11 . The biomass according to  claim 10 , wherein the thioesterase is a codon optimized for the C 1  metabolizing non-photosynthetic microorganism. 
     
     
         12 . The biomass according to  claim 10 , wherein the malonyl CoA-acyl carrier protein transacylase is a codon optimized  E. coli  fabD. 
     
     
         13 . The biomass according to  claim 10 , wherein the acetyl-CoA carboxylase is a codon optimized  E. coli  accA, accB, accC, accD, or any combination thereof. 
     
     
         14 . The biomass according to  claim 10 , wherein the C 1  metabolizing microorganism further comprises a mutation that minimizes or eliminates fatty acid-CoA ligase activity. 
     
     
         15 . The biomass according to  claim 1 , wherein the biomass comprises a spent media supernatant composition from a culture of a recombinant C 1  metabolizing non-photosynthetic microorganism and wherein the δ 13 C of the spent media composition is less than about −30‰. 
     
     
         16 . The biomass according to  claim 1 , wherein an oil composition is extracted or concentrated from the spent media composition.