Angelini Luciana G, Campeol Elisabetta, Tozzi Sabrina, Gilbert Kerry G, Cooke David T, John Philip
Dipartimento di Agronomia e Gestione dell'Agroecosistema, Università di Pisa, via S.Michele degli Scalzi 2, 56100 Pisa, Italy.
Biotechnol Prog. 2003 Nov-Dec;19(6):1792-7. doi: 10.1021/bp0300218.
A method to quantify the indigo precursor indican (indoxyl-beta-D-glucoside) in Polygonum tinctorium L. has been developed. Plant material was extracted in deionized water, and indican was identified and quantified using high performance liquid chromatography (HPLC) coupled to an evaporative light scattering detector (ELSD). Results confirmed that with this method it is possible to measure indican content in a short time, obtaining reliable and reproducible data. Using this method, leaf indican content was quantified every 15 days during the growing season (from May to October) in P. tinctorium crops grown in a field experiment in Central Italy. Results showed that indican increased along the growing season until flowering and was positively affected by photosynthetic active radiation (PAR). Indican is naturally hydrolyzed by native beta-glucosidase to indoxyl and glucose, the indoxyl yielding indigo. The activity of two enzymes, sweet almond beta-glucosidase and Novarom G preparation, were compared with P. tinctorium native beta-glucosidase to evaluate indigo production. Results showed that the ability to promote indigo formation increased as follows: almond beta-glucosidase <or= Novarom G.
已开发出一种定量蓼蓝中靛蓝前体靛苷（吲哚酚-β-D-葡萄糖苷）的方法。将植物材料用去离子水提取，然后使用配备蒸发光散射检测器（ELSD）的高效液相色谱（HPLC）对靛苷进行鉴定和定量。结果证实，采用该方法能够在短时间内测定靛苷含量，获得可靠且可重复的数据。利用此方法，在意大利中部进行的田间试验中，对蓼蓝作物生长季节（5月至10月）每隔15天的叶片靛苷含量进行了定量。结果表明，靛苷在生长季节一直增加直至开花，且受到光合有效辐射（PAR）的正向影响。靛苷会被天然的β-葡萄糖苷酶自然水解为吲哚酚和葡萄糖，吲哚酚进而生成靛蓝。将甜杏仁β-葡萄糖苷酶和Novarom G制剂这两种酶的活性与蓼蓝天然β-葡萄糖苷酶进行比较，以评估靛蓝的产量。结果表明，促进靛蓝形成的能力按以下顺序增强：杏仁β-葡萄糖苷酶≤Novarom G。
