Structure and function of a novel fungal enzyme, lactonohydrolase, and its application to optical resolution of pantolactone

新型真菌酶内酯水解酶的结构和功能及其在泛内酯光学拆分中的应用

• 批准号: 06453167
• 负责人: SHIMIZU Sakayu
• 金额: $ 5.06万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06453167/
• 关键词: Pantolactone; Optical resolution; Lactonase; Fusarium oxysporum; High-mannose sugarchain

A fungal lactonohydrolase catalyzes the stereospecific hydrolysis of the intramolecular ester bond of D-pantoyl lactone and is useful for optical resolution of racemic pantoyl lactone. High activity of this stereospecific hydrolysis reaction was found in several filamentous fungi belonging to the genera Fusarium, Gibberella and Cylindrocarpon through the screening in a variety of microorganisms. Fusarium oxysporum AKU 3702 showed high productivity of the enzyme and the cells containing the enzyme could be used repeatedly for this hydrolysis reaction. On incubation with the mycelia of this fungus, which had been cultivated in 3% glycerol, 0.5% Polypepton, 0.5% yeast extract and 0.5% corn steep liquor, pH 6.0,46.0% of the recemic pantoyl lactone (700 mg/ml) was hydrolyzed and the optical purity of the pantoic acid formed was 96% enantiomeric excess for the D-isomer.

真菌内酯水解酶催化D-泛酰内酯的分子内酯键的立体特异性水解，并可用于外消旋泛酰内酯的光学拆分。通过对多种微生物的筛选，发现在镰刀菌属、赤霉属和刺果霉属的几种丝状真菌中，这种立体专一性水解反应具有较高的活性。尖孢镰刀菌AKU 3702表现出较高的酶产率，并且含有该酶的细胞可重复用于该水解反应。在用该真菌的菌丝体（已在3%甘油、0.5%多肽、0.5%酵母提取物和0.5%玉米浆中培养，pH 6.0）孵育时，46.0%的外消旋泛解酸内酯（700 mg/ml）被水解，并且形成的泛解酸的光学纯度为96%，对于D-异构体而言，对映体过量。

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