Biomass estimation during macro-scale solid-state fermentation of basidiomycetes using established and novel approaches
Biomass estimation during macro-scale solid-state fermentation of basidiomycetes using established and novel approaches
复制标题
DOI：
10.1007/s00449-015-1372-0
复制
发表时间：
2015-07-01
影响因子：
3.8
通讯作者：
Bley, Thomas
中科院分区：
文献类型：
--
作者：
Steudler, Susanne;Bley, Thomas
固态发酵（SSF）已经在食品生产中使用了数千年，由于该过程的稳健性和使用木质纤维素作为底物的可能性，它非常适合于担子菌的培养。担子菌产生多种酶和各种初级和次级代谢物，其中许多具有生物技术潜力。真菌生物量的量化对于表征SSF等过程中的生长动力学至关重要。在SSF中，真菌生长到基质中，并将其作为营养来源。因此，直接测定生物量是不可能的，必须采用间接方法。在本研究中，我们比较了在一个新开发的实验室反应器（工作体积为10 L）中测定担子菌毛毡菌SSF过程中真菌生物量的11种方法。这些方法是基于测量六种细胞特异性成分（麦角甾醇、葡萄糖胺、核酸、真菌核数、蛋白质和基因组DNA）的水平和生物活性（呼吸、木质素和纤维素水解酶的活性以及液体中的葡萄糖和蛋白质含量）的估计。从结果的重现性和可信性、所需时间和资源、可能的影响因素和基质效应等方面对方法进行评价。最可靠的生物量估计是从麦角甾醇含量、核数和呼吸量的测量中获得的。因此，这三种方法被认为是最适合过程控制和建模的。
Solid-state fermentation (SSF) has been utilised in food production for millennia and is well suited for the cultivation of basidiomycetes, due to the robustness of the process and the possibility of using lignocellulose as the substrate. Basidiomycetes produce diverse enzymes and various primary and secondary metabolites, many of which have biotechnological potential. The quantification of the fungal biomass present is essential for the characterisation of growth kinetics in processes such as SSF. In SSF, fungi grow into the substrate and use it as a nutrient source. Therefore, direct biomass determination is not possible and indirect methods have to be employed. In the presented study, we compared 11 methods for quantifying fungal biomass during SSF of the basidiomycete Trametes hirsuta in a newly developed laboratory reactor (working volume 10 L). The methods were based on measuring the levels of six cell-specific components (ergosterol, glucosamine, nucleic acids, number of fungal nuclei, protein and genomic DNA) and estimations of biological activity (respiration, activities of lignolytic and cellulolytic enzymes, and the glucose and protein contents of the liquid). The methods were evaluated with regards to reproducibility and plausibility of the results, time and resource requirements, possible influential factors, and matrix effects. The most reliable biomass estimates were obtained from measurements of ergosterol content, number of nuclei, and respiration. Thus, these three methods were deemed most suitable for process control and modelling.