The methylotrophic yeast Komagataella phaffii is a eukaryotic microorganism capable of utilizing methanol as sole carbon source, having been widely employed for protein and metabolite production.However, its endogenous methanol utilization efficiency requires further improvement.This study overexpressed encoding genes of key enzymes in the non-oxidative branch of the pentose phosphate pathway (PPP), including transketolase (tkt), ribulose-5-phosphate-3-epimerase (rpe), and fructose-1,6-bisphosphatase (fba), to enhance methanol assimilation.Results showed that tkt-GS115 exhibited a 1.7% increase in OD600, with a maximum specific growth rate (μ) of 0.23 h-1.Compared to the control strain ΔFLDH, fba-ΔFLDH entered the growth phase 18 h earlier, achieving a similar μ of 0.23 h-1.Notably, rpe-ΔFDH and fba-ΔFDH rescued the growth defect of the parental strain ΔFDH in methanol minimal medium.Specifically, rpe-ΔFDH displayed significant growth potential, reaching a maximum OD600of 6.97, while fba-ΔFDH achieved a remarkably high μ of 0.36 h-1.Methanol utilization efficiency varied among the recombinant strains, including tkt-GS115, tkt-ΔFLDH, and fba-ΔFLDH showed improvements of 0.017, 0.007, and 0.034, respectively.This study demonstrates that reinforcing three key genes in the PPP non-oxidative branch effectively enhances methanol utilization efficiency in K.phaffii recombinant strains.
ZANG Xiu
,
YANG Yankun
,
BAI Zhonghu
. Regulating non-oxidative pentose phosphate pathway genes to improve methanol utility of Komagataella phaffii[J]. Food and Fermentation Industries, 2025
, 51(20)
: 1
-8
.
DOI: 10.13995/j.cnki.11-1802/ts.043026
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