Kickenweiz, T.; Glieder, A.; Wu, J. C., Construction of a cellulose-metabolizing Komagataella phaffii (Pichia pastoris) by co-expressing glucanases and β-glucosidase. Applied Microbiology and Biotechnology 2018, 102 (3), 1297-1306.
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.
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Funding Info:
This work was financially supported by the Science and Engineering Research Council (SERC) of the Agency for Science, Technology and Research (A*STAR), Singapore (SERC grant no 1526004159, ICES/15-175A02) and by the A*STAR Research Attachment Programme Scholarship from the A*STAR Graduate Academy.