College of Enology, Northwest A&F University, Yangling, Shaanxi, China.
Shaanxi Engineering Research Center for Viti-Viniculture, Yangling, Shaanxi, China.
J Food Sci. 2021 Jun;86(6):2327-2345. doi: 10.1111/1750-3841.15741. Epub 2021 Apr 30.
β-glucosidase is a pivotal enzyme that hydrolyzes bound volatile aromatic compounds. However, the activity of β-glucosidase in winemaking and the mechanism by which it affects the flavor and taste of wines have not been fully investigated. In this study, we profiled the characteristics of β-glucosidase derived from wine-related yeasts isolated from different wine-making regions in China, and analyzed the enzyme activity from different parts of the cells under aerobic and anaerobic conditions. A total of 56 strains of wine-related yeasts producing β-glucosidases were screened using the YNB-C medium (YNB 6.7 g L , cellobiose 5 g L , pH 5.0). We found that strain Clavispora lusitaniae C117 produced the highest enzyme activity (152.39 µmol pNP ml h ). In most strains, β-glucosidase were located in whole cells (periplasmic space) and permeabilized cells (intracellular). The non-Saccharomyces species had the highest enzymatic activity in a strain-dependent manner. Under aerobic conditions, C. lusitaniae C117, Hanseniaspora guilliermondii A27-3-4, Metschnikowia pulcherrima F-1-6, and Pichia anomala C84 had the highest β-glucosidase activity. We further investigated the β-glucosidase activity during the wine fermentation and the effects of sugar, pH, temperature, and ethanol on the enzyme activities of P. anomala C84 and commercial Saccharomyces yeast strains RC212 and VL1. The presence of fructose, glucose, and sucrose strongly inhibited enzyme activity. Similarly, low pH and low temperature inhibited the activity of β-glucosidase, whereas ethanol promoted enzyme activity. Our findings provide a theoretical basis on understanding the different yeast characteristics of β-glucosidase and their potential application for further improving wine aroma complexity.
β-葡萄糖苷酶是一种关键的酶，可水解结合的挥发性芳香化合物。然而，β-葡萄糖苷酶在酿酒中的活性以及它影响葡萄酒风味和口感的机制尚未得到充分研究。在本研究中，我们对从中国不同酿酒地区分离得到的与葡萄酒相关的酵母中提取的β-葡萄糖苷酶的特性进行了分析，并在有氧和无氧条件下分析了细胞不同部位的酶活性。使用 YNB-C 培养基（YNB 6.7 g/L，纤维二糖 5 g/L，pH 5.0）筛选出 56 株产生β-葡萄糖苷酶的与葡萄酒相关的酵母。我们发现，酿酒酵母 C117 产生的酶活性最高（152.39 µmol pNP ml h）。在大多数菌株中，β-葡萄糖苷酶位于整个细胞（周质空间）和渗透化细胞（细胞内）中。非酿酒酵母属的菌株在依赖于菌株的方式下具有最高的酶活性。在有氧条件下，C. lusitaniae C117、汉逊酵母 A27-3-4、酒香酵母 F-1-6 和异常毕赤酵母 C84 具有最高的β-葡萄糖苷酶活性。我们进一步研究了葡萄酒发酵过程中的β-葡萄糖苷酶活性，以及糖、pH 值、温度和乙醇对异常毕赤酵母 C84 和商业酿酒酵母 RC212 和 VL1 菌株的酶活性的影响。果糖、葡萄糖和蔗糖的存在强烈抑制了酶的活性。同样，低 pH 值和低温也抑制了β-葡萄糖苷酶的活性，而乙醇则促进了酶的活性。我们的研究结果为了解不同酵母β-葡萄糖苷酶的特性及其在进一步提高葡萄酒香气复杂性方面的潜在应用提供了理论依据。
