Multi-strategy Metabolic Engineering of Talaromyces pinophilus for α-Amylase Production from Lignocellulosic Biomass.

Authors

Jing Zeng, Jianjun Guo, Shuaiwen Zhang, Ya Li, Siyuan Yue, Peng Li, Junhui Nie, Tong Wang, Cheng Zhang, Lin Yuan

Published in

Journal of industrial microbiology & biotechnology. Aug 18, 2026. Epub Aug 18, 2026.

Abstract

Filamentous fungi are important hosts for industrial enzyme production. Growing demand for α-amylase has increased reliance on food-derived carbon substrates, necessitating fungal strains that efficiently utilize non-grain biomass. In this study, Talaromyces pinophilus Y117 was metabolically engineered to produce α-amylase from lignocellulosic biomass. A strong cellobiohydrolase I gene (cbh1) promoter (Pcbh1Tru) was identified to drive expression. Multiple rounds of multi-locus integration of the α-amylase gene were performed using homologous multi-copy genomic sequences as recombination arms with a Cre/loxP-based recyclable selection system, yielding the multi-copy strain Tp4, which achieved 4124.5 U/mL α-amylase activity in shake-flask fermentation with corncob powder as the sole carbon source. To minimize enzyme degradation, the protease gene 8538 was deleted using the Cre/lox2272 system, generating Tp4Δp. This strain showed a 50% increase in shake-flask α-amylase activity (6208.4 U/mL). In 3-L bioreactor cultivation, Tp4Δp exhibited excellent production performance, achieving 26 712.2 U/mL α-amylase activity. When corncob powder was used as the sole substrate, the cellulose and hemicellulose degradation rates reached 90.00% and 70.01%, respectively, and the enzyme yield reached 213 697.5 U per gram of corncob powder. This engineered strain demonstrates strong potential for industrial applications. The synthesis-degradation synergistic optimization strategy provides a practical approach for engineering filamentous fungal cell factories to produce enzymes directly from lignocellulosic biomass.

PMID:
42610752
Bibliographic data and abstract were imported from PubMed on 18 Aug 2026.

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