Efficient methanol assimilation was crucial for methanol-based biomanufacturing of high-valued products.Given the low methanol utility in native Komagataella phaffii cells,we disrupted the genes encoding formaldehyde ...Efficient methanol assimilation was crucial for methanol-based biomanufacturing of high-valued products.Given the low methanol utility in native Komagataella phaffii cells,we disrupted the genes encoding formaldehyde dehydrogenase and formate dehydrogenase and incorporated heterologous RuMP into K.phaffii to drive more flux into central metabolic pathways.We also performed transcriptome analysis to evaluate the metabolic impact of this genetic modification.The results showed that the biomass ofΔfldh mutant strain was 8.3%higher than that of the wild type control strain.The ratio of biomass accumulation from methanol was respectively 5.754 and 6.209 in WT andΔfldh.Recombinant RuMP-Δfdh and hps-Δfdh rescued the unlivingΔfdh and were able to growth in medium with methanol as sole carbon source.In addition,the transcription revealed the impact of disrupted fldh or fdh on TCA,PPP and the respiratory chain.Our results suggested that inactivation of fldh and expressing RuMP were beneficial for methanol utility.It was fdh rather than fldh that was indispensable for K.phaffii cell growth.This study provided new insights into how to reprogram K.phaffii to enhance its methanol assimilation rate,also the theoretical bases for the mechanism underlying the better methanol digestion.展开更多
基金supported by the National Natural Science Foundation of China(32370054)the national first-class discipline program of Light Industry Technology and Engineering(LITE2018-24).
文摘Efficient methanol assimilation was crucial for methanol-based biomanufacturing of high-valued products.Given the low methanol utility in native Komagataella phaffii cells,we disrupted the genes encoding formaldehyde dehydrogenase and formate dehydrogenase and incorporated heterologous RuMP into K.phaffii to drive more flux into central metabolic pathways.We also performed transcriptome analysis to evaluate the metabolic impact of this genetic modification.The results showed that the biomass ofΔfldh mutant strain was 8.3%higher than that of the wild type control strain.The ratio of biomass accumulation from methanol was respectively 5.754 and 6.209 in WT andΔfldh.Recombinant RuMP-Δfdh and hps-Δfdh rescued the unlivingΔfdh and were able to growth in medium with methanol as sole carbon source.In addition,the transcription revealed the impact of disrupted fldh or fdh on TCA,PPP and the respiratory chain.Our results suggested that inactivation of fldh and expressing RuMP were beneficial for methanol utility.It was fdh rather than fldh that was indispensable for K.phaffii cell growth.This study provided new insights into how to reprogram K.phaffii to enhance its methanol assimilation rate,also the theoretical bases for the mechanism underlying the better methanol digestion.
