Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, China.
Appl Biochem Biotechnol. 2022 Jul;194(7):3016-3030. doi: 10.1007/s12010-022-03891-y. Epub 2022 Mar 25.
Enzymatic hydrolysis using β-mannanase and α-galactosidase is necessary to produce low molecular weight galactomannan (LMW-GM) from galactomannans (GM) in the leguminous seeds. In this study, different ratios of avicel and melibiose were used as the inductors (carbon sources) for Trichoderma reesei to metabolize the enzyme cocktail containing β-mannanase and α-galactosidase using one-pot fermentation technology. The obtained enzyme cocktail was used to efficiently produce LMW-GM from GM in Sesbania cannabina seeds. Results showed that 15 g/L avicel and 10 g/L melibiose were the best carbon sources to prepare enzyme cocktail containing β-mannanase and α-galactosidase with activities of 3.69 ± 0.27 U/mL and 0.51 ± 0.02 U/mL, respectively. Specifically, melibiose could effectively induce the metabolite product of α-galactosidase by T. reesei, which showed good performance in degrading the galactose substituent from GM backbone. The degradation of galactose alleviated the spatial site-blocking effect for enzymatic hydrolysis by β-mannanase and improved the yield of LMW-GM. This research can lay the foundation for the industrial technology amplification of LMW-GM production for further application.
利用β-甘露聚糖酶和α-半乳糖苷酶进行酶解,是从豆科种子中的半乳甘露聚糖(GM)中生产低分子量半乳甘露聚糖(LMW-GM)所必需的。在这项研究中,使用不同比例的微晶纤维素和棉子糖作为诱导物(碳源),用于里氏木霉代谢含有β-甘露聚糖酶和α-半乳糖苷酶的酶混合物,采用一锅发酵技术。所得的酶混合物用于从田菁种子中的 GM 中高效生产 LMW-GM。结果表明,15 g/L 微晶纤维素和 10 g/L 棉子糖是制备含有β-甘露聚糖酶和α-半乳糖苷酶的酶混合物的最佳碳源,其酶活分别为 3.69±0.27 U/mL 和 0.51±0.02 U/mL。具体来说,棉子糖可以有效地诱导里氏木霉产生α-半乳糖苷酶的代谢产物,这对半乳糖取代物从 GM 主链上的降解表现出良好的性能。半乳糖的降解减轻了β-甘露聚糖酶酶解的空间位阻效应,提高了 LMW-GM 的产量。这项研究可以为 LMW-GM 生产的工业技术放大奠定基础,以进一步应用。