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Expression of enzymes in yeast for lignocellulose derived oligomer CBP

A lignocellulose and xylanase technology, applied in the direction of enzymes, biofuels, biochemical equipment and methods, etc., can solve the problem of expensive external enzyme addition

Inactive Publication Date: 2015-07-22
LALLEMAND HUNGARY LIQUIDITY MANAGEMENT LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, adding enzymes externally is expensive

Method used

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  • Expression of enzymes in yeast for lignocellulose derived oligomer CBP
  • Expression of enzymes in yeast for lignocellulose derived oligomer CBP
  • Expression of enzymes in yeast for lignocellulose derived oligomer CBP

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0393] Production of M3799 and M3059, robust and efficient xylose-utilizing Saccharomyces cerevisiae strains

[0394] The strains described in U.S. Application No. 61 / 557,971, filed November 10, 2011, and International Application No. PCT / US2012 / 064457, filed November 9, 2012 (both of which are incorporated herein by reference) M2874, which is a xylose-utilizing derivative of the robust S. cerevisiae strain M2390, was engineered to improve its performance on concentrated C5-containing liquors derived from hardwoods. To achieve this goal, the strains were subjected to two forms of continuous culture (chemostat and pH auxostat), with the results as figure 2 described in.

[0395] The soluble sugars produced in the process were washed out of the solids by hardwood chips pretreated at a severity of 3.8, in a laboratory autoclave at 121 °C with 0.5% H 2 SO 4 Hydrolyze them for 3 hours, followed by 15M NH 4 OH to adjust the pH to 5.8 to produce a C5-containing liquor. The solu...

Embodiment 2

[0400] Characterization of accessory cellulase expression and activity

[0401] Soluble oligomers extracted from hardwood after pretreatment consist of various sugars linked in multiple ways. Linkages require different types of enzymes to hydrolyze them into their constituent monosaccharides. Figure 5 Several oligomers presumed to be present in hardwood-derived oligomers based on a review of the literature related to the composition of hardwoods are presented (e.g. Wilfor (2005) Wood Sci Tech; Teleman (2003) Carbohydrate Res; Shallom (2003) Curr Op Biotech ; Spanikova (2006) FEBS Letters; Rowell, RM. Handbook of Wood Chemistry and Composites. London: Taylor & Francis (2005)), and the general types of enzymes expected to hydrolyze these types of bonds.

[0402] Extracted forms of oligomers, especially xylooligomers, are noteworthy as potent inhibitors of cellulase during cellulose hydrolysis (Qing, Q., Yang, B., Wyman, C.E. Xylooligomers are strong inhibitors of cellulose hy...

Embodiment 3

[0508] Generation of engineered strains of Saccharomyces cerevisiae for enzyme expression and integrated bioprocessing of hardwood-derived soluble oligomers

[0509] As described above, a group of enzymes was discovered by screening various enzyme types and several amino acid sequences for each active type capable of hydrolyzing soluble oligomers isolated from lignocellulose with very high efficiency. The next set of enzymes selected for strain engineering was based on these data.

[0510] Table 13. Enzymes selected for CBP strain construction

[0511]

[0512] Using U.S. Application No. 61 / 557,971, filed November 10, 2011, International Application No. PCT / US2012 / 064457, and International Publication No. WO2011 / 140386, filed November 9, 2012 (these applications and publications are adopted Several strains were generated by the method described in (incorporated herein by reference) for the integration of genetic element islands into S. cerevisiae. The strains produced to ...

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Abstract

The present invention provides a multi-component enzyme system that hydrolyzes hemicellulose oligomers from hardwood which can be expressed, for example, in yeast such as Saccharomyces cerevisiae. In some embodiments, this invention provides for the engineering of a series of biocatalysts combining the expression and secretion of components of this enzymatic system with robust, rapid xylose utilization, and ethanol fermentation under industrially relevant process conditions for consolidated bioprocessing. In some embodiments, the invention utilizes co-cultures of strains that can achieve significantly improved performance due to the incorporation of additional enzymes in the fermentation system.

Description

[0001] Reference Sequence Listing [0002] The contents of the Sequence Listing filed with this application are hereby incorporated by reference in their entirety. [0003] Statement Regarding Federally Funded Research and Development [0004] This invention was made in part by the US Government and Department of Energy under Award No. DE-FC36-08GO18103. This invention was also supported in part by Bioenergy Science Center Oak Ridge National Laboratory, a US Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research, under contract DE-PS02-06ER64304. The US Government has certain rights in this invention. Background of the invention [0005] Energy conversion, utilization and access constitute many of the great challenges of our time, including those related to sustainable development, environmental quality, security and poverty. New applications of emerging technologies are needed to address these challenges. Biotechnol...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N15/81
CPCY02E50/16C12P19/02C12P19/14C12N9/18C12P7/10C12N9/248C12Y301/01072C12N9/2402C12N9/2437C12N9/2445C12N9/2465C12N9/2471C12N9/2482C12N9/2491C12N9/2494C12Y301/01006C12Y302/01021C12Y302/01037C12Y302/01072C12Y302/01131C12Y302/01139Y02E50/10C12P3/00C12P7/16C12P7/28C12P7/52C12P7/56
Inventor J·E·麦克布莱德E·威斯瓦尔I·施卡雷H·许N·索恩格恩H·H·豪尔E·斯通豪斯
Owner LALLEMAND HUNGARY LIQUIDITY MANAGEMENT LLC
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