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J Biosci Bioeng ; 128(1): 33-38, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-30711353

RESUMEN

In this work, we analyzed several genes participating in the rearrangement pathway for xylulose 5-phosphate (Xu5P) in the methylotrophic yeast Pichia pastoris (syn. Komagataella phaffii). P. pastoris has two set of genes for fructose-1,6-bisphosphate aldolase (FBA1 and FBA2) and transaldolase (TAL1 and TAL2), although there are single-copy genes for fructose-1,6-bisphosphatase (FBP1) and transketolase (TKL1), respectively. Expressions of FBP1 and TAL2 were upregulated by non-fermentative carbon sources, especially methanol was the best inducer for them, and FBA2 was induced only by methanol. On the other hand, FBA1, TAL1 and TKL1 showed constitutive expression. Strain fbp1Δ showed severe growth defect on methanol and non-fermentable carbon sources, and growth rate of strain fba2Δ in methanol medium was slightly decreased. Moreover, Fba2p and Tal2p possessed peroxisome targeting signal type 1 (PTS1), and EGFP-Fba2p and EGFP-Tal2p were found to be localized in peroxisomes. From these findings, it was suggested that Fba2p, Fbp1p and Tal2p participate in the rearrangement pathway for Xu5P in peroxisomes, and that the altered Calvin cycle and non-oxidative pentose phosphate pathway involving Tal2p function in a complementary manner.


Asunto(s)
Fructosa-Bifosfato Aldolasa/genética , Metanol/metabolismo , Pentosafosfatos/metabolismo , Pichia , Transaldolasa/genética , Fructosa-Bifosfato Aldolasa/metabolismo , Regulación Enzimológica de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Prueba de Complementación Genética , Redes y Vías Metabólicas/genética , Peroxisomas/genética , Peroxisomas/metabolismo , Pichia/enzimología , Pichia/genética , Pichia/crecimiento & desarrollo , Pichia/metabolismo , Saccharomyces cerevisiae/metabolismo , Transaldolasa/metabolismo , Transcetolasa/genética , Transcetolasa/metabolismo
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