背景:在过去的二十年里,杂交一直是用于酿造和酿酒的优质酵母的强大工具。新型杂种主要使用至少一种酿酒酵母亲本构建。然而,对用于其他目的的杂种知之甚少,例如有针对性的香料生产,例如,2-苯基乙醇(2-PE)。2-PE,一种广泛用于食品中的芳香化合物,化妆品,和制药工业,由于其毒性,在生物技术生产中提出了挑战。因此,为了提高生产率和对2-PE的耐受性,各种策略,如诱变和基因工程被广泛探索,以改善酵母菌株。虽然生物技术的努力主要集中在2-PE生产的酿酒酵母上,其他酵母属物种及其杂种仍未得到充分描述。
结果:为了解决这个差距,在这项研究中,我们分析了一种新的种间酵母杂种,II/6,来自S.uvarum和S.kudriavzevii的父母,就2-PE生物转化和对其高浓度的抗性而言,将其与亲本菌株进行比较。在本研究中使用两种已知的2-PE生物转化介质和三种不同的温度来确定最佳条件。在72小时分批培养中,II/6杂种的最大值为2.36±0.03g/L2-PE,比亲本菌株的生产率高2-20倍。我们的兴趣不仅在于确定杂种的生产率是否提高,还在于评估其对高2-PE滴度的敏感性是否也得到缓解。结果表明,杂种对毒性产物的抗性明显高于原始菌株。
结论:所进行的实验证实了杂交是一种有前途的用于修饰酵母菌株的方法。因此,2-PE产量和对其抑制作用的耐受性都可以提高。此外,这种策略允许获取非转基因菌株,减轻与其他立法要求或消费者对生产者的接受问题有关的担忧。获得的发现有可能为将来开发实用的解决方案做出贡献。
BACKGROUND: Over the last two decades, hybridization has been a powerful tool used to construct superior yeast for brewing and winemaking. Novel hybrids were primarily constructed using at least one Saccharomyces cerevisiae parent. However, little is known about hybrids used for other purposes, such as targeted flavor production, for example, 2-phenylethanol (2-PE). 2-PE, an aromatic compound widely utilised in the food, cosmetic, and pharmaceutical industries, presents challenges in biotechnological production due to its toxic nature. Consequently, to enhance productivity and tolerance to 2-PE, various strategies such as mutagenesis and genetic engineering are extensively explored to improved yeast strains. While biotechnological efforts have predominantly focused on S. cerevisiae for 2-PE production, other Saccharomyces species and their hybrids remain insufficiently described.
RESULTS: To address this gap, in this study, we analysed a new interspecies yeast hybrid, II/6, derived from S. uvarum and S. kudriavzevii parents, in terms of 2-PE bioconversion and resistance to its high concentration, comparing it with the parental strains. Two known media for 2-PE biotransformation and three different temperatures were used during this study to determine optimal conditions. In 72 h batch cultures, the II/6 hybrid achieved a maximum of 2.36 ± 0.03 g/L 2-PE, which was 2-20 times higher than the productivity of the parental strains. Our interest lay not only in determining whether the hybrid improved in productivity but also in assessing whether its susceptibility to high 2-PE titers was also mitigated. The results showed that the hybrid exhibited significantly greater resistance to the toxic product than the original strains.
CONCLUSIONS: The conducted experiments have confirmed that hybridization is a promising method for modifying yeast strains. As a result, both 2-PE production yield and tolerance to its inhibitory effects can be increased. Furthermore, this strategy allows for the acquisition of non-GMO strains, alleviating concerns related to additional legislative requirements or consumer acceptance issues for producers. The findings obtained have the potential to contribute to the development of practical solutions in the future.