Selection of suitably non-repressing carbon sources for expression of alcohol oxidase isozyme promoters in the methylotrophic yeast Pichia methanolica

Nakagawa, Tomoyuki; Wakayama, Keishi; Hayakawa, T.

doi:10.1016/j.jbiosc.2014.12.007

Cited by 4 publications

(3 citation statements)

References 19 publications

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“…4A). These results are consistent with those of previous reports (Nakagawa et al ., 2004, 2010a, 2015). Moreover, intracellular levels of thiamine pyrophosphate (TPP) and flavin adenine dinucleotide (FAD), which are co‐enzymes for AOD isozymes and DAS, respectively, in L‐MeOH‐grown cells were significantly increased compared with Glc‐grown cells ( P < 0.05; Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The yeast can rapidly adapt its carbon source to shift from glucose to methanol without medium replacement and followed by biomass production (Raymond et al, 1998). Furthermore, O. methanolica has two strong AOD promoters (Nakagawa et al, 2006(Nakagawa et al, , 2015, and the non-homologous recombination for the expression cassettes performs effectively in O. methanolica (Hiep et al, 1993). However, to realize highly productive methanol biomass conversions, a fermentation system that can work efficiently under high methanol conditions, particularly to produce commodity chemicals, is required.…”

Section: Introductionmentioning

confidence: 99%

“…The yeast can rapidly adapt its carbon source to shift from glucose to methanol without medium replacement and followed by biomass production (Raymond et al ., 1998). Furthermore, O. methanolica has two strong AOD promoters (Nakagawa et al ., 2006, 2015), and the non‐homologous recombination for the expression cassettes performs effectively in O. methanolica (Hiep et al ., 1993).…”

Section: Introductionmentioning

confidence: 99%

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Metabolic regulation adapting to high methanol environment in the methylotrophic yeast Ogataea methanolica

Cai

Doi²,

Shimada

et al. 2021

Microbial Biotechnology

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Since methylotrophic yeasts such as Ogataea methanolica can use methanol as a sole carbon feedstock, they could be applied to produce valuable products from methanol, a next-generation energy source synthesized from natural gases, using genetic engineering tools. In this study, metabolite profiling of O. methanolica was conducted under glucose (Glc) and low and high methanol (L-and H-MeOH) conditions to show the adaptation mechanism to a H-MeOH environment. The yeast strain responded not only to the presence of methanol but also to its concentration based on the growth condition. Under H-MeOH conditions, O. methanolica downregulated the methanol utilization, glycolytic pathway and alcohol oxidase (AOD) isozymes and dihydroxyacetone synthase (DAS) expression compared with L-MeOH-grown cells. However, levels of energy carriers, such as ATP, were maintained to support cell survival. In H-MeOH-grown cells, reactive oxygen species (ROS) levels were significantly elevated. Along with increasing ROS levels, ROS scavenging system expression was significantly increased in H-MeOH-grown cells. Thus, we concluded that formaldehyde and H 2 O 2 , which are products of methanol oxidation by AOD isozymes in the peroxisome, are overproduced in H-MeOH-grown cells, and excessive ROS derived from these cells is generated in the cytosol, resulting in upregulation of the antioxidant system and downregulation of the methanol-utilizing pathway to suppress overproduction of toxic intermediates.

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Section: Resultsmentioning

confidence: 99%