State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China.
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China.
Food Chem. 2020 Aug 1;320:126635. doi: 10.1016/j.foodchem.2020.126635. Epub 2020 Mar 19.
In pear, sucrose was mainly distributed in vacuole; and the alternation of sucrose abundance was associated the change of vacuolar invertase (VI) activity during fruit storage. However, the molecular mechanism beneath such phenomenon has not been clarified until recently. For this, a combination of metabolite, enzyme activity, transcriptome, quantitative real-time PCR (qRT-PCR), bioinformation, subcellular localization, and transient overexpression assay was conducted in this study to identify the acid invertase 1 (PbrAc-Inv1) and invertase inhibitor 5 (PbrII5) involved in sucrose degradation during 'Housui' pear storage. Both PbrAc-Inv1 and PbrII5 were located in vacuolar membrane. PbrAc-Inv1 could accelerate sucrose degradation; on the other hand, PbrII5 could bind with PbrAc-Inv1 to form a inactive complex, downregulate the VI activity, and suppressed sucrose decomposition. Based on Bio-layer interferometry (BLI) result after domain substitution, the domain on the left of catalytic 'WEC-P/V-D' box in PbrAc-Inv1 might played a key role in its interaction with PbrII5.
在梨中,蔗糖主要分布在液泡中;在果实贮藏过程中,蔗糖含量的变化与液泡转化酶(VI)活性的变化有关。然而,直到最近,这种现象背后的分子机制仍不清楚。为此,本研究结合代谢物、酶活性、转录组、定量实时 PCR(qRT-PCR)、生物信息学、亚细胞定位和瞬时过表达分析,鉴定了参与‘Housui’梨贮藏过程中蔗糖降解的酸性转化酶 1(PbrAc-Inv1)和转化酶抑制剂 5(PbrII5)。PbrAc-Inv1 和 PbrII5 均定位于液泡膜上。PbrAc-Inv1 可加速蔗糖降解;另一方面,PbrII5 可与 PbrAc-Inv1 结合形成无活性复合物,下调 VI 活性,抑制蔗糖分解。基于结构域替换后的生物层干涉(BLI)结果,PbrAc-Inv1 中催化‘WEC-P/V-D’框左侧的结构域可能在其与 PbrII5 的相互作用中发挥关键作用。