Construction of a cellulose-metabolizing Komagataella phaffii (Pichia pastoris) by co-expressing glucanases and β-glucosidase

Thomas Kickenweiz, Anton Glieder, Jin Chuan Wu

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Abstract

Cellulose is a highly available and renewable carbon source in nature. However, it cannot be directly metabolized by most microbes including Komagataella phaffii (formerly Pichia pastoris), which is a frequently employed host for heterologous protein expression and production of high-value compounds. A K. phaffii strain was engineered that constitutively co-expresses an endoglucanase and a β-glucosidase both from Aspergillus niger and an exoglucanase from Trichoderma reesei under the control of bidirectional promoters. This engineered strain was able to grow on cellobiose and carboxymethyl cellulose (CMC) but not on Avicel. However, the detected release of cellobiose from Avicel by using the produced mixture of endoglucanase and exoglucanase as well as the released glucose from Avicel by using the produced mixture of all three cellulases at 50 °C indicated the production of exoglucanase under the liquid culture conditions. The successful expression of three cellulases in K. phaffii demonstrated the feasibility to enable K. phaffii to directly use cellulose as a carbon source for producing recombinant proteins or other high-value compounds.

Originalspracheenglisch
Seiten (von - bis)1297-1306
FachzeitschriftApplied Microbiology and Biotechnology
Jahrgang102
Ausgabenummer3
DOIs
PublikationsstatusVeröffentlicht - 2018

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Glucosidases
Pichia
Cellulose
Cellobiose
Cellulases
Cellulase
Carbon
Carboxymethylcellulose Sodium
Trichoderma
Aspergillus niger
Recombinant Proteins
Glucose
Proteins

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    Construction of a cellulose-metabolizing Komagataella phaffii (Pichia pastoris) by co-expressing glucanases and β-glucosidase. / Kickenweiz, Thomas; Glieder, Anton; Wu, Jin Chuan.

    in: Applied Microbiology and Biotechnology, Jahrgang 102, Nr. 3, 2018, S. 1297-1306.

    Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

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    AB - Cellulose is a highly available and renewable carbon source in nature. However, it cannot be directly metabolized by most microbes including Komagataella phaffii (formerly Pichia pastoris), which is a frequently employed host for heterologous protein expression and production of high-value compounds. A K. phaffii strain was engineered that constitutively co-expresses an endoglucanase and a β-glucosidase both from Aspergillus niger and an exoglucanase from Trichoderma reesei under the control of bidirectional promoters. This engineered strain was able to grow on cellobiose and carboxymethyl cellulose (CMC) but not on Avicel. However, the detected release of cellobiose from Avicel by using the produced mixture of endoglucanase and exoglucanase as well as the released glucose from Avicel by using the produced mixture of all three cellulases at 50 °C indicated the production of exoglucanase under the liquid culture conditions. The successful expression of three cellulases in K. phaffii demonstrated the feasibility to enable K. phaffii to directly use cellulose as a carbon source for producing recombinant proteins or other high-value compounds.

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