产黑色素酵母 Sporidiobolus ruineniae A45.2 共合成没食子酸和一种新型细胞结合单宁酶。
Co-production of gallic acid and a novel cell-associated tannase by a pigment-producing yeast, Sporidiobolus ruineniae A45.2.
机构信息
Division of Biochemistry and Biochemical Technology, Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
Research Center for Multidisciplinary Approaches to Miang, Chiang Mai University, Chiang Mai, 50200, Thailand.
出版信息
Microb Cell Fact. 2020 Apr 25;19(1):95. doi: 10.1186/s12934-020-01353-w.
BACKGROUND
Gallic acid has received a significant amount of interest for its biological properties. Thus, there have been recent attempts to apply this substance in various industries and in particular the feed industry. As opposed to yeasts, fungi and bacteria and their tannases have been well documented for their potential bioconversion and specifically for the biotransformation of tannic acid to gallic acid. In this research, Sporidiobolus ruineniae A45.2 is introduced as a newly pigment-producing and tannase-producing yeast that has gained great interest for its use as an additive in animal feed. However, there is a lack of information on the efficacy of gallic acid production from tannic acid and the relevant tannase properties. The objective of this research study is to optimize the medium composition and conditions for the co-production of gallic acid from tannic acid and tannase with a focus on developing an integrated production strategy for its application as a feed additive.
RESULTS
Tannase produced by S. ruineniae A45.2 has been classified as a cell-associated tannase (CAT). Co-production of gallic acid obtained from tannic acid and CAT by S. ruineniae A45.2 was optimized using response surface methodology and then validated with the synthesis of 11.2 g/L gallic acid from 12.3 g/L tannic acid and the production of 31.1 mU/mL CAT after 48 h of cultivation in a 1-L stirred tank fermenter. Tannase was isolated from the cell wall, purified and characterized in comparison with its native form (CAT). The purified enzyme (PT) revealed the same range of pH and temperature optima (pH 7) as CAT but was distinctively less stable. Specifically, CAT was stable at up to 70 °C for 60 min, and active under its optimal conditions (40 °C) at up to 8 runs.
CONCLUSION
Co-production of gallic acid and CAT is considered an integrated and green production strategy. S. ruineniae biomass could be promoted as an alternative source of carotenoids and tannase. Thus, the biomass, in combination with gallic acid that was formed in the fermentation medium, could be directly used as a feed additive. On the other hand, gallic acid could be isolated and purified for food and pharmaceutical applications. This paper is the first of its kind to report that the CAT obtained from yeast can be resistant to high temperatures of up to 70 °C.
背景
没食子酸因其生物特性而受到广泛关注。因此，最近人们试图将这种物质应用于各个行业，特别是饲料行业。与酵母、真菌和细菌及其单宁酶不同，它们的单宁酶已被充分证明具有潜在的生物转化能力，特别是将单宁酸转化为没食子酸的生物转化能力。在这项研究中，介绍了一种新型产色素和产单宁酶的酵母 Sporidiobolus ruineniae A45.2，由于其可用作动物饲料添加剂，因此备受关注。然而，关于从单宁酸生产没食子酸和相关单宁酶特性的信息还很缺乏。本研究的目的是优化从单宁酸和单宁酶共生产没食子酸的培养基组成和条件，重点是开发一种综合生产策略，将其作为饲料添加剂应用。
结果
S. ruineniae A45.2 产生的单宁酶已被归类为细胞结合型单宁酶（CAT）。使用响应面法对 S. ruineniae A45.2 从单宁酸共生产没食子酸进行了优化，然后在 1 L 搅拌罐发酵罐中培养 48 小时后，从 12.3 g/L 单宁酸合成 11.2 g/L 没食子酸和生产 31.1 mU/mL CAT 进行了验证。将单宁酶从细胞壁中分离出来，并与天然形式（CAT）进行了比较和纯化。与 CAT 相比，纯化酶（PT）的最适 pH 和温度范围相同（pH 7），但稳定性明显较差。具体而言，CAT 在 60 分钟内可在高达 70°C 的温度下稳定，并且在其最佳条件（40°C）下可在 8 次运行中保持活性。
结论
共生产没食子酸和 CAT 被认为是一种综合的绿色生产策略。S. ruineniae 生物量可作为类胡萝卜素和单宁酶的替代来源。因此，发酵培养基中形成的没食子酸和生物质可直接用作饲料添加剂。另一方面，没食子酸可被分离和纯化用于食品和制药应用。本文首次报道，从酵母中获得的 CAT 可以耐受高达 70°C 的高温。
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