State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
J Sci Food Agric. 2011 Oct;91(13):2443-8. doi: 10.1002/jsfa.4485. Epub 2011 May 27.
Momordica charantia (MC) is used in many Asian countries as a traditional functional food and medicine. Polypeptide-P, a 166 amino acid (AA) polypeptide isolated from MC seeds, has been reported to show hypoglycaemic effects in patients with type I or type II diabetes. The AA sequence of this peptide has been determined, but its gene sequence has yet to be published.
In this study a gene-cloning strategy was employed to obtain the polypeptide-P gene sequence using degenerate reverse transcription polymer chain reaction and genome-walking methods. A complete 498 bp sequence encoding the polypeptide-P protein was cloned from MC seeds. Subsequent assays of the bioactivity of the expressed recombinant protein revealed that it had significant hypoglycaemic activity in alloxan-induced diabetic mice. This result suggests that recombinant polypeptide-P has hypoglycaemic effects.
This is the first report of cloning and expression of the MC polypeptide-P gene. The cloned gene could be helpful for exploring the mechanisms of polypeptide-P gene expression and regulation in MC. Furthermore, this gene could be used as a potential tool both for screening MC varieties with high hypoglycaemically active substance content and for breeding new varieties of MC with high economic value, which could in turn be beneficial to farmers.
苦瓜(Momordica charantia)在许多亚洲国家被用作传统的功能性食品和药物。从苦瓜种子中分离出的 166 个氨基酸(AA)多肽——多肽-P,已被报道对 I 型或 II 型糖尿病患者具有降血糖作用。该肽的 AA 序列已被确定,但它的基因序列尚未公布。
在这项研究中,我们采用基因克隆策略,使用简并逆转录聚合酶链反应和基因组步移方法获得多肽-P 基因序列。从苦瓜种子中克隆出一个完整的编码多肽-P 蛋白的 498bp 序列。随后对表达的重组蛋白的生物活性进行检测,结果表明该蛋白在四氧嘧啶诱导的糖尿病小鼠中具有显著的降血糖活性。这一结果表明重组多肽-P 具有降血糖作用。
这是首次报道苦瓜多肽-P 基因的克隆和表达。克隆的基因有助于探索苦瓜中多肽-P 基因表达和调控的机制。此外,该基因可作为一种潜在的工具,用于筛选具有高降血糖活性物质含量的苦瓜品种,培育具有高经济价值的苦瓜新品种,从而有利于农民。