| Title |
Recombinant Expression of Trichoderma reesei Cel61A in Pichia pastoris: Optimizing Yield and N-terminal Processing
|
|---|---|
| Published in |
Molecular Biotechnology, August 2015
|
| DOI | 10.1007/s12033-015-9887-9 |
| Pubmed ID | |
| Authors |
Magali Tanghe, Barbara Danneels, Andrea Camattari, Anton Glieder, Isabel Vandenberghe, Bart Devreese, Ingeborg Stals, Tom Desmet |
| Abstract |
The auxiliary activity family 9 (AA9, formerly GH61) harbors a recently discovered group of oxidative enzymes that boost cellulose degradation. Indeed, these lytic polysaccharide monooxygenases (LPMOs) are able to disrupt the crystalline structure of cellulose, thereby facilitating the work of hydrolytic enzymes involved in biomass degradation. Since these enzymes require an N-terminal histidine residue for activity, their recombinant production as secreted protein is not straightforward. We here report the expression optimization of Trichoderma reesei Cel61A (TrCel61A) in the host Pichia pastoris. The use of the native TrCel61A secretion signal instead of the alpha-mating factor from Saccharomyces cerevisiae was found to be crucial, not only to obtain high protein yields (>400 mg/L during fermentation) but also to enable the correct processing of the N-terminus. Furthermore, the LPMO activity of the enzyme is demonstrated here for the first time, based on its degradation profile of a cellulosic substrate. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| France | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 110 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Japan | 1 | <1% |
| Unknown | 109 | 99% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 28 | 25% |
| Researcher | 23 | 21% |
| Student > Master | 17 | 15% |
| Student > Bachelor | 7 | 6% |
| Student > Doctoral Student | 4 | 4% |
| Other | 11 | 10% |
| Unknown | 20 | 18% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 34 | 31% |
| Agricultural and Biological Sciences | 31 | 28% |
| Engineering | 4 | 4% |
| Chemistry | 3 | 3% |
| Chemical Engineering | 2 | 2% |
| Other | 7 | 6% |
| Unknown | 29 | 26% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 20 August 2015.
All research outputs
#20,288,585
of 22,824,164 outputs
Outputs from Molecular Biotechnology
#804
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Outputs of similar age
#223,252
of 266,176 outputs
Outputs of similar age from Molecular Biotechnology
#5
of 8 outputs
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