Enhanced cellodextrin metabolism

A technology of cellodextrin and dextrin, applied in the direction of transferase, peptide, enzyme, etc., can solve problems such as determination

Inactive Publication Date: 2014-02-19
RGT UNIV OF CALIFORNIA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these engineered strains may not ferment glucose with optimal metabolism because the strain's endogenous system for detecting and responding to the presence of glucose depends on extracellular glucose levels

Method used

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  • Enhanced cellodextrin metabolism
  • Enhanced cellodextrin metabolism
  • Enhanced cellodextrin metabolism

Examples

Experimental program
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preparation example Construction

[0419] Other aspects of the invention relate to the preparation of host cells containing recombinant cellodextrin transporter, recombinant cellodextrin phosphorylase, recombinant β-glucosidase, recombinant glucose phosphomutase or recombinant hexokinase one or more. In some embodiments, the host cell may further comprise one or more glucose response genes of the invention, one or more pentose transporters of the invention, and / or genes involved in pentose utilization, of one or more recombinases. These host cells can be used to degrade cellodextrins and produce carbohydrates or carbohydrate derivatives from cellodextrins.

[0420] The method of producing and culturing the host cell of the present invention may comprise introducing or transforming an expression vector containing the recombinant polynucleotide into the host cell. Those of ordinary skill in the art are aware of these methods for transforming expression vectors into host cells. For example, one method of transf...

Embodiment 1

[0463] introduction

[0464] Engineering microbes to convert sugars found in plant cell walls into fuels and other chemicals has garnered considerable attention. Plant cell walls are composed of cellulose (a polymer of glucose), hemicellulose (a heterogeneous polymer of pentoses, hexoses, and sugar acids), and lignin (a heterogeneous phenolic polymer). Plant cell walls are abundant in agriculture and municipal waste, and are dedicated energy crops. The yeast, Saccharomyces cerevisiae, is a favored platform for these engineering projects because of its stability, ease of genetic manipulation, and ability to undergo high-carbon transitions. In addition to this, Saccharomyces cerevisiae has numerous deficiencies including inability to naturally ferment pentose sugars, sensitivity to solvents and sensitivity to inhibitory compounds found in decomposed plant material.

[0465] Another drawback is that Saccharomyces cerevisiae cannot naturally ferment cellodextrins, such as cellob...

Embodiment 2

[0567] This example describes the identification of conserved motifs for cellodextrin phosphorylases and the identification of additional cellodextrin phosphorylases.

[0568] The first round of PSI-BLAST was run with Clostridium thermocellum (BAA22081.1), Acidovibrio cellulolyticus (ZP_07328763.1) and Clostridium lentocellum (YP_004310865.1) cellodextrin phosphorylase amino acid sequences as simultaneous input. From the results of the first round, all query results annotated as "cellodextrin phosphorylase" were used as simultaneous input for the second round of PSI-BLAST.

[0569] From the results of the second round, all sequences annotated as "cellodextrin phosphorylase" were used to generate a multiple sequence alignment by T-COFFEE.

[0570] Use multiple sequence alignment as the PRATT server ( http: / / web.expasy.org / pratt / ) to identify the highest scoring motif. The motif is shown in PROSITE format:

[0571] G-x(2)-[FY]-x-N-[AGS]-x-[AS]-W-[APS]-V-[IL]-[AS]-x(2)-A-x(2...

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Abstract

The present disclosure relates to host cells containing two or more of a recombinant cellodextrin transporter, a recombinant cellodextrin phosphorylase, a recombinant ss-glucosidase, a recombinant phosphoglucomutase, or a recombinant hexokinase; and to methods of using such cells for degrading cellodextrin, for producing hydrocarbons or hydrocarbon derivatives from cellodextrin, and for reducing ATP consumption during glucose utilization.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application 61 / 440,305, filed February 7, 2011, and US Provisional Application 61 / 566,548, filed December 2, 2011, both of which are hereby incorporated by reference in their entirety. [0003] Sequence listing submitted as an ASCII text file [0004] The contents of the ASCII text file submission are hereby incorporated herein by reference: Computer Readable Form (CRF) Sequence Listing (File Name: 677792001340SeqList.txt, Date of Record: February 6, 2012, Size: 1209KB). technical field [0005] The present invention relates to methods and compositions for the degradation of cellodextrins and for the production of hydrocarbons and hydrocarbon derivatives. Background technique [0006] Biofuels have been intensively investigated due to rising concerns about energy security, sustainability, and global climate change (Lynd et al., Science, 1991). The bioconversion of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/08
CPCC12N9/2445C12N9/1205Y02E50/16C12P7/08C12P19/02C12Y504/02002C12Y204/0102C12P7/10C12Y302/01021C12Y204/01049Y02E50/10C07K14/395C12N9/1051C12N9/90C12P19/18
Inventor 詹姆斯·H·多德纳-凯特陈勇秀乔纳森·M·加拉兹卡析辰河
Owner RGT UNIV OF CALIFORNIA
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