Gottfried Wilhelm Leibniz Universität Hannover, Institut für Lebensmittelchemie, Callinstraße 5, 30167, Hannover, Germany.
Bioprocess Biosyst Eng. 2018 Sep;41(9):1391-1401. doi: 10.1007/s00449-018-1966-4. Epub 2018 Jun 12.
A glycosidase of the basidiomycete Bjerkandera adusta (BadGluc) was found in screenings to possess a strong decolorizing ability towards malvidin-3-galactoside, an anthocyanin abundant in various berry fruits. The BadGluc was purified from the culture supernatant via FPLC, and the corresponding gene was identified which showed low similarity to other characterized glucosidases. Scanning the primary sequence with PROSITE no active site motif was detected. Eventually, a specific 18 aa consensus pattern was identified manually. The active site motif possessed an undescribed sequence which was only found in a few hypothetical proteins. The corresponding gene was cloned and expressed in Pichia pastoris GS115 yielding activities up to 100 U/L using 4-nitrophenyl-β-d-glucopyranoside (pNPG) as substrate. The enzyme possessed a good temperature (70% after 1 h at 50°C) and pH stability (70% between pH 2 and 7.5), and preferably catalysed the hydrolysis of delphinidin-3-glucoside and cyanidin-3-glucoside, regardless of the position of the terminal Hexa-His tag. This novel glucosidase worked in aqueous solution as well as on pre-stained fabrics making it the first known candidate anthocyanase for applications in the detergent and food industries.
从土壤真菌 Bjerkandera adusta 中发现的一种糖苷酶（BadGluc）具有很强的矢车菊素-3-半乳糖苷（在各种浆果中含量丰富的一种花青素）的脱色能力。BadGluc 通过 FPLC 从培养上清液中纯化，鉴定了相应的基因，该基因与其他特征糖苷酶的相似度较低。用 PROSITE 扫描一级序列未检测到活性位点基序。最终，通过手动鉴定出一个特定的 18 个氨基酸共识模式。活性位点基序具有一个未描述的序列，仅在少数假设蛋白中发现。相应的基因被克隆并在毕赤酵母 GS115 中表达，使用 4-硝基苯基-β-d-吡喃葡萄糖苷（pNPG）作为底物时，酶活最高可达 100 U/L。该酶具有良好的温度（50°C 下 1 小时后保留 70%的活性）和 pH 稳定性（在 pH 2 到 7.5 之间保留 70%的活性），并且无论末端六组组氨酸标签的位置如何，都优先催化飞燕草素-3-葡萄糖苷和矢车菊素-3-葡萄糖苷的水解。这种新型糖苷酶在水溶液和预染色织物中均能发挥作用，使其成为用于洗涤剂和食品工业的首个已知的花青素酶候选酶。
