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Transcription factors for the production of cellulose degrading enzymes

A transcription factor, a technology encoding transcription factors, applied in the field of cellulose degradation

Inactive Publication Date: 2014-07-09
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this strategy has greatly reduced the cost of enzyme production, the resulting strains have hundreds of mutations

Method used

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  • Transcription factors for the production of cellulose degrading enzymes
  • Transcription factors for the production of cellulose degrading enzymes
  • Transcription factors for the production of cellulose degrading enzymes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0315] Example 1: Induction of cellulolytic enzymes in wild-type Neurospora crassa

[0316] To better understand how filamentous fungi sense and respond to cellulose in their environment, next-generation RNA sequencing was used to profile genome-wide mRNA abundance in Neurospora crassa. For these experiments, cultures grown for 16 hours in sucrose minimal medium (SMM; linear growth phase) and then transferred from SMM to cellulose-only carbon source medium (CMM; cellulose minimal medium); RNA samples were taken at 30 min, 1 hr, 2 hr and 4 hr after transfer from SMM to CMM and compared to cultures transferred to SMM at the same time points. Typical expression patterns of genes known to be involved in cellulose degradation are shown in Figure 1A middle. The expression level of genes in this subset increased by about 1 order of magnitude within 30 min after transfer. Transcript abundance remained constant for about 1 hour before increasing several orders of magnitude betwee...

Embodiment 2

[0320] Example 2: Necessary Regulators for Cellulose Degradation

[0321] To identify transcription factors required for cellulose degradation, we screened ~200 N. crassa transcription factor deletion sets for mutants with deletions grown on cellulose. Two mutants with severe growth defects on cellulose but normal growth on sucrose were identified. For cellulose degradation regulators 1 and 2, the corresponding genes NCU07705 and NCU08042 were tentatively named cdr-1 and cdr-2, respectively. However, it was later discovered that the prefix "cdr" was previously used for genes related to cadmium resistance in Neurospora crassa. Therefore, for cellulose degradation regulators 1 and 2, the genes NCU07705 and NCU08042 were renamed clr-1 and clr-2, respectively. It should be noted that although some figures may refer to cdr-1 and cdr-2, the descriptions of these figures in Examples 2-4 below refer to cdr-1 as clr-1 and cdr-2 as clr -2.

[0322] Deletion mutants of clr-1 and cl...

Embodiment 3

[0327] Example 3: Phylogenetic Analysis

[0328] A phylogenetic analysis of clr-1 and clr-2 was performed to gain insight into the evolutionary history of the two genes. The maximum likelihood phylogenetic tree of clr-1 and clr-2 homologues substantially recapitulates the previous fungal tree ( Figure 6 ). Trees generated using Bayesian inference also showed consistent trees (Fig. 9). The similarity between the two trees provides support for the idea that CLR-1 and CLR-2 may have co-evolved and the hypothesis that they may function together as a hybrid.

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Abstract

Provided herein are methods and compositions for increasing the production of one or more cellulases from a fungal host cell. The disclosure is based, on the surprising discovery that mis-expression of the transcriptional regulator clr-2 in a filamentous fungal cell was able to induce expression of cellulase genes under non-inducing or starvation conditions, resulting in increased secretion of cellulases from the cell. Advantageously, mis-expression of the transcription factor clr-2 in a filamentous fungal cell cultured in the absence of cellulose or cellobiose results in increased secretion of cellulases. The disclosure relates inter alia to a method of degrading cellulose-containing material, to a method of increasing the production of one or more cellulases from a fungal cell and to a method of reducing the viscosity of a pretreated biomass material, by contacting pretreated biomass material with a fungal host cell containing at least one recombinant nucleic acid encoding clr-2 or a related transcription factor.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 61 / 510,466, filed July 21, 2011, which is hereby incorporated by reference in its entirety. [0003] Submission of a sequence listing as an ASCII text file [0004] The following submission as an ASCII text file is hereby incorporated by reference in its entirety: Sequence Listing in Computer Readable Form (CRF) (File Name: 677792002140SeqList.txt, Data Recorded: Jul 23, 2012, Size: 978KB). field of invention [0005] The present disclosure relates to the degradation of cellulose. In particular, the disclosure relates to polypeptides involved in cellular response to cellulose, as well as related nucleotides and compositions. The present disclosure also relates to methods and uses of polypeptides, nucleotides and compositions thereof in relation to cellular responses to cellulose. Background technique [0006] Liquid fuels derived from biomass ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C07K14/37
CPCC12P19/14C07K14/37C12N15/80C12P19/02
Inventor 田朝光T·肖克N·L·格拉斯S·科拉代蒂J·克雷格
Owner RGT UNIV OF CALIFORNIA
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