University of Belgrade - Institute of Chemistry, Technology and Metallurgy - National Institute of the Republic of Serbia, Department of Chemistry, Njegoševa 12, 1100 Belgrade, Republic of Serbia.
Insitute of Pesticide and Environmental Protection, Banatska 31b, 11000 Belgrade-Zemun, Republic of Serbia.
Food Res Int. 2022 Oct;160:111755. doi: 10.1016/j.foodres.2022.111755. Epub 2022 Jul 29.
Production of fructooligosaccharides (FOS) is a trending topic due to their prebiotic effect becoming increasingly important for the modern human diet. The most suitable process for FOS production is the one using fungal inulinases. Introduction of new fungal inulinase producers and their implementation in production of inulinase enzymes is therefore gaining interest. This study provides a new approach to FOS synthesis by fungal enzyme complex without prior separation of any specific enzyme. Inulinase enzyme complexes could be used for the synthesis of FOS in two possible ways - hydrolysis of inulin (FOS) and transfructosylation process of sucrose (FOS), as demonstrated here. Depending on the fungal growth inducing substrate, a variety of inulinase enzyme complexes was obtained - one of which was most successful in production of FOS and another one of FOS. Substrates derived from crops: triticale, wheat bran, Jerusalem artichoke and Aspergillus welwitschiae isolate, previously proven as safe for use in food, were utilized for production of inulinase enzyme cocktails. The highest FOS production was obtained by enzyme complex rich in β-fructofuranosidase, while the highest FOS production was obtained by enzyme complex rich in endoinulinase. Both FOS and FOS showed antioxidant potential according to ABTS and ORAC, which classifies them as a suitable additive in functional food. Simultaneous zymographic detection of inulinase enzymes, which could contribute to expansion of the knowledge on fungal enzymes, was developed and applied here. It demonstrated the presence of different inulinase isoforms depending on fungal growth substrate. These findings, which rely on the innate ability of fungi to co-produce all inulinases from a cocktail, could be useful as a new, easy approach to FOS production by fungal enzymes without their separation and purification, contributing to cheaper and faster production processes.
由于其益生元作用对于现代人类饮食变得越来越重要，因此果糖低聚糖 (FOS) 的生产是一个热门话题。生产 FOS 最适合的方法是使用真菌菊粉酶。因此，引入新的真菌菊粉酶生产菌并将其应用于菊粉酶的生产正引起人们的兴趣。本研究提供了一种新的方法，通过真菌酶复合物在没有预先分离任何特定酶的情况下合成 FOS。可以通过两种可能的方式使用菊粉酶酶复合物合成 FOS - 菊粉 (FOS) 的水解和蔗糖 (FOS) 的转果糖基化过程，如这里所示。根据真菌生长诱导底物的不同，可以获得各种菊粉酶酶复合物 - 其中一种在 FOS 的生产中最成功，另一种在 FOS 的生产中最成功。来源于作物的底物：黑小麦、麦麸、洋姜和 Aspergillus welwitschiae 分离株，以前被证明可安全用于食品，被用于生产菊粉酶酶混合物。富含β-果聚糖酶的酶复合物获得了最高的 FOS 产量，而富含内切菊粉酶的酶复合物获得了最高的 FOS 产量。根据 ABTS 和 ORAC，FOS 和 FOS 都显示出抗氧化潜力，这将它们归类为功能性食品中的合适添加剂。同时开发并应用了同时进行菊粉酶酶的同工酶检测，这有助于扩展对真菌酶的了解。它证明了存在不同的菊粉酶同工酶，这取决于真菌生长的底物。这些发现依赖于真菌从混合物中共产生所有菊粉酶的固有能力，可作为一种新的、简单的方法，无需分离和纯化真菌酶即可生产 FOS，有助于降低成本和加快生产过程。
