Peroxisomal Fba2p and Tal2p complementally function in the rearrangement pathway for xylulose 5-phosphate in the methylotrophic yeast Pichia pastoris
Peroxisomal Fba2p and Tal2p complementally function in the rearrangement pathway for xylulose 5-phosphate in the methylotrophic yeast Pichia pastoris
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DOI：
10.1016/j.jbiosc.2019.01.008
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发表时间：
2019-07-01
影响因子：
2.8
通讯作者：
Nakagawa, Tomoyuki
中科院分区：
文献类型：
--
作者：
Fukuoka, Hirotaka;Kawase, Takato;Nakagawa, Tomoyuki
在这项工作中，我们分析了几个基因参与重排途径的木酮糖5-磷酸（Xu 5 P）在甲醇营养酵母巴斯德毕赤酵母（syn. Komagataella phaffi）。巴斯德毕赤酵母具有果糖-1，6-二磷酸醛缩酶（FBA 1和FBA 2）和转醛醇酶（TAL 1和TAL 2）的两组基因，尽管分别存在果糖-1，6-二磷酸酶（FBP 1）和转酮醇酶（TKL 1）的单拷贝基因。非发酵碳源对FBP 1和TAL 2的表达有促进作用，其中甲醇对FBP 1和TAL 2的诱导作用最强，而FBA 2仅受甲醇的诱导。另一方面，FBA 1、TAL 1和TKL 1显示组成型表达。菌株fbp 1 Delta在甲醇和非发酵碳源上表现出严重的生长缺陷，而菌株fba 2 Delta在甲醇培养基中的生长速率略有下降。Fba 2 p和Tal 2 p具有过氧化物酶体靶向信号类型1（PTS 1），EGFP-Fba 2 p和EGFP-Tal 2 p定位于过氧化物酶体。这些结果表明，Fba 2 p、Fbp 1 p和Tal 2 p参与了Xu 5 P在过氧化物酶体中的重排途径，Tal 2 p参与的卡尔文循环改变和非氧化戊糖磷酸途径以互补的方式发挥作用。(C)2019年，日本生物技术学会。All rights reserved.
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 phaffi). 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 fbp1 Delta showed severe growth defect on methanol and non-fermentable carbon sources, and growth rate of strain fba2 Delta 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. (C) 2019, The Society for Biotechnology, Japan. All rights reserved.