The potential applications of Pichia pastoris for methanol bioconversion are extensive. In order to elucidate the mechanism of methanol metabolism in P. pastoris, four peroxisomal isoenzymes (i.e., Rpe1-2, Rki1-2, Tal1-2, and Fba1-2.) involved in methanol utilization (MUT) pathway were investigated. Their roles in methanol metabolism were verified by constructing single-gene knockout mutants and quadruple-gene knockout mutant. Moreover, cytoplasmic isozymes and peroxisomal isozymes were complemented in quadruple-gene knockout mutant, respectively. The results showed that the growth of quadruple-gene knockout mutant was severely inhibited in methanol medium, and the Fba1-2 deficiency had the greatest effect on the growth phenotype. At the same time, Fba1-2 complementation could rescue the biomass production of quadruple-gene knockout mutant most effectively. These results further confirm that Fba1-2 is a crucial enzyme for the methanol assimilation and highlight the significance of peroxisome localization of assimilation pathway. Additionally, by comparing the growth phenotypes of SHB17 single-gene knockout mutant and TAL1-2 single-gene knockout mutant, it is further confirmed that the altered Calvin cycle is the main pathway of regenerating XU5P in methanol metabolism. This study can provide theoretical basis for optimizing the MUT pathway in P. pastoris.
ZOU Xiaoyi
,
WANG Shijie
,
YANG Yankun
,
BAI Zhonghu
. The role of four peroxisomal isoenzymes on methanol metabolism in Pichia pastoris[J]. Food and Fermentation Industries, 2024
, 50(3)
: 9
-15
.
DOI: 10.13995/j.cnki.11-1802/ts.035425
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