Host cells and methods for isobutanol production

A technology of host cells and recombinant host cells, which is applied in the field of recombinant host cells and can solve problems such as high price and non-environmental protection

Active Publication Date: 2018-04-17
BUTAMAXTM ADVANCED BIOFUELS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods use starting materials derived from petrochemicals and are generally expensive and not environmentally friendly

Method used

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  • Host cells and methods for isobutanol production
  • Host cells and methods for isobutanol production
  • Host cells and methods for isobutanol production

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0208] The biological diversity screening of KARI-encoding genes from various bacterial and fungal species is described in Example 1, which shows a suitable KARI for isobutanol production. Using the present invention, those skilled in the art should easily be able to identify other suitable polypeptides with KARI activity.

[0209] The sequences disclosed herein and available in the art can be used to identify sequences of other polynucleotides, genes and / or polypeptides from the literature and bioinformatics databases well known to the skilled person. For example, such sequences can be identified by BLAST searches of publicly available databases using the polynucleotide or polypeptide sequences provided herein. In such methods, the identity can be based on the Clustal W alignment method, which uses GAP PENALTY=10, GAP LENGTHPENALTY=0.1, and the default parameters of the Gonnet 250 series of protein weight matrix.

[0210] In addition, the polynucleotide or polypeptide sequences d...

example

[0460] The present invention will be further defined in the following examples. It should be understood that, although these examples show preferred embodiments of the present invention, they are only given in an illustrative manner. Through the above discussion and these examples, those skilled in the art can determine the essential characteristics of the present invention, and without departing from the spirit and scope of the present invention, can make various changes and modifications to the present invention to adapt to various applications and conditions. .

[0461] General method :

[0462] The standard recombinant DNA and molecular cloning techniques used in the examples are well known in the art and are described in: Sambrook, J., Fritsch, EF and Maniatis, T., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press , Cold Spring Harbor, NY, 1989, TJSilhavy, ML; Bennan and LWEnquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold ...

example 2

[0490] Screening of KARI diversity collections for isobutanol production

[0491] The various KARI genes are evaluated based on their "effective yield" in yeast. The effective yield is determined after a certain period of growth (for example, 48h) under progressive oxygen-limited conditions. Yeast biomass at 1OD 600 The yeast cells are equivalent to 0.3g / L.

[0492] Yeast isobutanol pathway strains carrying different KARI genes were inoculated into 10 ml SEG-Ura, His medium with 0.2% glucose and 0.2% ethanol, and grown aerobic at 30°C overnight to about 2OD. Centrifuge the culture and resuspend part of the cells in SEG-Ura, His (2% glucose, 1% ethanol) in a 125ml shake flask to reach an initial OD of 0.4 in a total volume of 25ml 600 . The shake flask is closed with a screw-tight solid plastic cap, and the culture is grown in the flask with the lowest air and oxygen exchange with the external environment under the condition of progressive oxygen limitation. After 48 hours of in...

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Abstract

Provided herein are recombinant yeast host cells that produce isobutanol and methods of their use. The provided yeast host cell comprises: isobutanol biosynthesis pathway and reduced or eliminated aldehyde dehydrogenase activity, reduced or eliminated at least one of the acetolactate reductase activity; or encoding has ketol acid reductoisomerase activity The heterologous polynucleotide of the polypeptide.

Description

[0001] Government licensing rights [0002] The present invention is supported by the US government under the contract number DE-AR0000006 awarded by the Department of Energy. The US government has certain rights in this invention. Technical field [0003] The present invention relates to a recombinant host cell and method for fermentation production of isobutanol. [0004] Cross references to related applications [0005] This patent application requires US patent applications 61 / 472,484 (filed on April 6, 2011), 61 / 467,261 (filed on March 24, 2011), 61 / 472,487 (filed on March 24, 2011) in accordance with 35U.SC§119(e). 2011 / 4 / 6), 61 / 467,271 (filed on March 24, 2011), 61 / 570,513 (filed on December 14, 2011), 61 / 467,249 (filed on March 24, 2011), The priority of 61 / 472,497 (filed on April 6, 2011) and 61 / 472,474 (filed on April 6, 2011), each of which is incorporated by reference in its entirety. [0006] Reference to the sequence table submitted electronically via EFS-WEB [0007] ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/61C12N15/53C12N15/63C12P7/16C12P7/52
CPCC12N1/38Y02E50/10C12P7/16C12N9/0006C12N15/52C12N15/63C12P7/52C12N15/815C12N15/81C12P7/42Y02P20/52
Inventor L.C.安东尼K.J.吉布森H.何L.L.黃D.P.奥基弗A.L.克鲁科伯格Y.李L.A.马格基奥-哈尔J.麦塞瓦恩M.J.内尔森R.帕特奈克S.C.罗斯曼
Owner BUTAMAXTM ADVANCED BIOFUELS
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