Glucoamylase is one of the most widely produced enzymes globally with diverse applications, such as in the fields of food, bioenergy, and papermaking. Microbially expressed glucoamylase serves as a potential additive in industrial alcohol fermentation to improve the utilization rate of starch. Komagataella phaffii is widely recognized for its capacity for high-density fermentation and efficient secretion of heterologous proteins, making it an ideal host for large-scale production under relatively simple fermentation conditions. However, the expression of glucoamylase in Komagataella phaffii currently faces challenges, including the potential toxicity of methanol as an inducer and overall low production yields. In this study, we first employed the improved transcription signal amplification device (iTSAD) in Komagataella phaffii to express the glucoamylase from Schizophyllum commune, achieving a titer of 3.31 g/L in a 5-L bioreactor along with a maximum glucoamylase activity of 5.2 U/mL. This study represents the first demonstration of high-efficiency methanol-free production of glucoamylase in Komagataella phaffii and provides strategies for the efficient expression and secretion of other industrial enzymes in Komagataella phaffii.
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Funding
Hubei Provincial Natural Science Foundation of China(2025AFD283)
Hubei Provincial Technology Innovation Programme Project of China(2025BCA006)
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Jiangnan University
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