Metabolic engineering of <i>Komagataella phaffii</i> for enhanced production of heterologous proteins and small molecules: key strategies and future directions

ABSTRACT Komagataella phaffii (syn. Pichia pastoris ) is an important host for the production of recombinant proteins and small molecules, often exhibiting superior performance compared to other heterologous hosts. With advancements in synthetic biology and computational biology, metabolic engineering has become a key strategy for achieving high-level heterologous proteins and small molecules in K. phaffii for industrial and commercial applications. In this review, we provide an overview of advances in K. phaffii metabolic engineering. We first discuss the development of the K. phaffii expression system. Subsequently, we analyze core optimization strategies, including metabolic flux control, metabolic dynamic control, application of system modeling, chassis cell optimization, and recombinant protein and small-molecule production, before finally summarizing the current challenges and future development directions.