Cyanobacterium sp. for production of compounds 机翻标题: 暂无翻译,请尝试点击翻译按钮。

公开号/公开日
WO2014100799 A3 2014-08-21 [WO2014100799]WO2014100799 A2 2014-06-26 [WO2014100799] / 2014-08-212014-06-26
申请号/申请日
2013WO-US77364 / 2013-12-23
发明人
WANG KUI;SHI TUO;PIVEN IRINA;INABA MASAMI;ULICZKA FRANK;KRAMER DAN;ENKE HEIKE;BAIER KERSTIN;FRIEDRICH ALEXANDRA;DUEHRING ULF;
申请人
ALGENOL BIOFUELS;
主分类号
IPC分类号
C12N-001/21C12N-015/63
摘要
(WO2014100799) A genetically enhanced cyanobacterial host cell, Cyanobacterium sp.  ABICyanoI, is disclosed.  The enhanced Cyanobacterium sp.  ABICyanoI produces a compound or compounds of interest.
机翻摘要
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地址
代理人
代理机构
;
优先权号
2012US-61741000 2012-12-21 2013US-61835294 2013-06-14
主权利要求
(WO2014100799) 1. A genetically enhanced. Cyanobacterium sp. host cell comprising at least one recombinant gene, wherein said recombinant gene encodes one protein selected from a group consisting of a. protein that is involved in a. biosyntheiie pathway for the production of a chemical compound or a marker protein. 45. The host cell of any of the pre vious claims for ethanol production, wherein the first recombinant gene encodes AdhE enzyme directly converting acetyl-CoA to ethanol. 46. A transformable Cyanobocte um sp. cell comprising an extracellular polymer layer (EPS) preheated with compounds selected from a group consisting of: N- aeetyleysteine, lysozynie, .beta.-galactosidase and combinations thereof. 63. A method for trans rming cyanobacteriai cells having an extracellular polymeric layer (EPS) comprising treating the cells with compounds selected from a group consisting of: N-acetylcysteine, lysozynie, and .beta.-galactosidase and combhiations thereof, before and/or during transformation with a recombinant UNA. 65. A plasmid vector suitable for transformation of Cyanohact rmm sp. ABICyano, comprising a recombinant gene, wherein said recombinant gene encodes at least one protein selected from a gr oup consisting of a protein thai is involved in a hiosynthetic pathway for the production of a chemical compound or a marker protein and an origin of replication suitable for replication in the Cyanohacteriwn ABICyanol. 77. Tlie plasmid vector of the preceding cl aims 73 to 76, wherein the first and/or second promoter is an inducible promoter and lias at least 90%, preferably 95% sequence identity or is identical to the promoters of SEQ ID NOs 9-4 i. 84. An isolated nucleic acid sequence having at least 95% identity to the sequence of the 6. 8 fcb plasmid (SEQ ID NO: 1). 85. An isolated Cyanobacteriwn sp. ABICyano 1 with the accession number PT A- 13311. 86. A method for introducing a recombinant nucleic acid sequence into a cyanobacterial cell with an extracellular polymeric layer (EPS) comprising the method steps of: A) subjecting the cyanobacterial cell to compounds increasing the permeability of the extracellular polymeric layer (EPS) and cell wall respectively of the cyanobacterial cell, and B) introducing said recombinant nucleic acid sequence into the cyanobacterial cell 88. A kit for producing a chemical compound via photosynthesis, comprising: - host cells of any of the c la ims 1 to 46 including the recombinant gene encoding one protein that is involved in a biosynthetic pathway for the production of said chemical compound, - a vessel for culturing the host cells, - means for illumination of the host cells. 93. A genetically enhanced ethanologenic copper inducible ABICyanol host cell. 95. A genetically enhanced ethanologenic ABICyan l host cell comprising adli genes that encode for alcohol dehydrogenase enzymes with catalytic properties that result in increased ethanol production relative to the ethanol production of alcohol dehydrogenase from Synechocystis sp. PCC 6803. 97. A genetically enhanced ethanologenic ABICyanol host cell comprising multiple copies of pdc genes, 99. The host cell of c laim 98 comprising plasmids selec ted irom the group consisting of #1792, #1743, #1744, #1749, #1751, #1817, #1818, #1728, and #1578. 100. A method for generating knockouts in the genome of ABICyanol comprising introducing a heterologous polynucleotide construct into a ABICyanol host cell wherein said heterologous polynucleotide construct comprises flanking regions of about 2 kbp or greater that are homologous to a. gene or gene portion in said genome of(')ABICyanol. 105. A method for producing ethanol using a genetically enhanced ethanologenic ABICyanol host cell comprising a plasmid selected from the group consisting of #1495, #1578, #1580, #1581, #1601, #1606, TK412 and TK411. 107. A genetically enhanced ethanologenic ABICyanol host cell comprising an ethanologenic cassette comprising adh and pdc genes that are under the control of endogenous metal inducible promoters. 115. A genetically enhanced ethanologenie ABICyanol host cell comprising a genomic knockout of a ycf37 gene. 116. A method for producing ethanol comprising knocking out a genomic ycf37 gene in an ethanologenie ABICyanol host cell. 117. A genetically enhanced ethanologenie Cyanobacterium sp. ABICyanol host cell. 140. A genetically enhanced ethanologenic Cy nobacteritmt sp.. ABICvanol host cell comprising a genetically enhanced plasmid derived from a plasmid that is endogenous to CycmobacteHum sp. ABICvanol, wherein said genetically enhanced plasmid comprises at least 50% of a 6.8 kfo plasmid endogenous to Cyanobacterium sp. ABICyanol consisting of SEQ ID NO: 1. 144. A genetically enhanced ethanologenic Cyanobacterium sp. ABICyanol host cell comprising Cyanobacteriwn sp. ABICyanol deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA-1331 1 comprising a genetically enhanced plasmid derived from a plasmid that is endogenous to Cyanobacterium sp. ABICyanol, and wherein said plasmid comprises a heterologous adh gene and a heterologous pdc gene, and wherein said adh gene is operably linked to a constitutive promoter, and wherein said pdc gene is operably linked to an inducible promoter, and wherein said Cyanobacterium sp. ABICyanol can tolerate salinities ranging up to about 35 practical salinity units, and can tolerate temperatures ranging up to about 55 degrees Celsius, and can tolerate dissolved oxygen concentrations ranging up to about 1000 .micro. .eta.o /L, and can tolerate ethanoi concentrations ranging up to about one percent for 16 weeks, and wherein production of ethanoi by said ethanologenic Cyanobacterium sp. ABICyanol host cell is from about 0.017 percent ethanoi (vol/vol) per OD[750] per day to about 0.031 percent ethanoi (vol/vol) per GD[750] per day. 147. A genetically enhanced etiianologenic Cyanobacterium sp. ABICyanoI host cell comprising Cyanobacterium sp. ABICyanoI deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA-1331 land a plasmid selected from the group consisting of TK293 (SEQ ID NO: 43), #1578 (SEQ ID NO: 48), and #1658. (SEQ ID NO: 72) and where m said Cyanobacterium sp. ABICyanoI can tolerate salinities ranging up to about 35 practical salinity units, and can tolerate temperatures ranging up to about 55 degrees Celsius, and can tolerate dissolved oxygen concentrations ranging up to about 1000 .micro.n o /L, and can tolerate ethanol concentrations ranging up to about one percent for 16 weeks, and wherein production of ethanol by said ethanologenic Cyanobacterium sp. ABICyanoI host cell i from about 0.024 percent ethanol (vol/vol) per OD[750] per day to about 0.049 percent ethanol (vol/vol) per OD[?]so per day. 148. A non-naturally occurring ethanologenic Cyanobacterium sp. organism. 152. A non-naturally occurring(') ethanologenic Cyanobacterium sp. ABICyanol herem said organism comprises a genetically enhanced plasmid derived from a plasmid that is endogenous to Cyanobacterium sp. ABICyanol selected from th group consisting of plasmids comprising SEQ ID NO. l and SEQ ID NO: 2. 156. A non-naturally occurring ethanologenic Cyanobacterium sp. ABICyanol organism comprising Cyanobacterium sp. ABICyanol deposited in the American Type Culture Collection (ATCC) under ATCC accession number TA- 33.pi., and further comprising an ethanologenic pathway and at least one exogenous nucleic acid encoding an ethanologenic pathway enzyme expressed in a sufficient amount to produce ethanol, said ethanologenic pathway comprising a pyruvate decarboxylase, and an alcohol dehydrogenase. 180. Wherein said constitutive promoter and said inducible promoter are endogenous to Cyanobacterium sp. ABICyanol. 184. A non-naturaily occurring Cyanobactetium sp. ABICyanol organism comprising Cyanobacterium sp. ABICyano deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA-13311 and a plasmid selected from the group consisting of TK293 (SEQ ID NO; 10) and #1578 (SEQ ID NO: 15), and wherein said Cyanobacterium sp. ABICyanol can tolerate salinities ranging up to about 35 practical salinity units, and can tolerate temperatures ranging up to about 55 degrees Celsius, and can tolerate dissolved oxygen concentrations ranging up to about 1000 .micro..eta. o /L, and can tolerate ethanol concentrations ranging up to about one percent for 16 weeks, and wherein production of ethanol by said ethanologenic Cyanobacterium sp. ABICyanol host cell is from about 0.031 percent ethanol (vol/vol) per QD[75]o per day to about 0.049 percent ethanol (vol/vol) per OD[750] er day. 85. A non-naturally occurring ethanologenic Cyanobacterium sp. organism derived from Cyanobacterimn sp. ABICyanol, wherein said organism comprises a genetically enhanced plasmid derived from a plasmid that is endogenous to Cyanobacterium sp. ABICyanol, and wherein said organism produces ethanol from about 0.002% (vol/vol) per day up to about 0.07% (vol/vol) per day. 205. A non-naturaliy occurring ethanologenic Cyanobacterimn sp. organism comprising Cyanobacterimn sp. ABICyanol deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA- 3311, and further comprising an ethanologenic pathway comprising one, two. or three exogenous nucleic acids each encoding an eihanologemc pathway enzyme expressed in a sufficient amount to produce ethanol. and wherein said ethanologenic pathway has been introduced into the chromosomal DNA of said Cyanobactetium sp. ABICyanol, and wherein said ethanologenic pathway comprises at least one pyruvate decarboxylase gene operably linked to a promoter and an alcohol dehydrogenase gene operably linked to a promoter. 209. A method for producing ethanoi comprising growing a non-naturally occurring ethanologenic Cyanobactermm sp. organism comprising Cyanobacterimn sp. ABICyano(')l deposited in the American Type Culture Collection (ATCC) under ATCC accession number PTA-13311, and further comprising an ethanologenic pathway comprising at least one exogenous nucleic acid encoding an ethanologenic pathway enzyme expressed in a sufficient amount for the organism to produce ethanoi, said ethanologenic pathway comprising a pyruvate decarboxylase; and an alcohol dehydrogenase. 215. Polynucleotide sequence of any of SEQ ID NOs 43, 61, 62, 66, 72, 73, 74, 82, 83, 84, 85, and 106. 4055. 4580 of SEQ ID NO: 43), P^ (nucleotides 2112-2680 of SEQ ID NO: 43), PpsbA (nucleotides 2112-2264 SEQ ID NO: 44), Ppeo (nucleotides 101-533 of SEQ ID NO: SOX (nucleotides 1169-1247 of SEQ ID NO: 55), [5! A] (nucleotides 480- 581 of SEQ ID NO: 56) and P[petE] (nucleotides 96-491 of SEQ ID NO; 46).
法律状态
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专利类型码
A3A2
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