Zhang Chuhan, Qin Lijun
Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), College of Life Sciences/Institute of Agro-Bioengineering, Guizhou University, 550025 Guiyang, China.
J Appl Microbiol. 2025 Jun 2;136(6). doi: 10.1093/jambio/lxaf007.
To determine the optimum conditions for extracting Eucommia ulmoides gum (EUG) from E. ulmoides leaves during fermentation by Coprinellus disseminatus. At the same time, the EUG characteristics were characterized.
The ability of C. disseminatus to secrete degrading enzymes, including enzyme activity and lignocellulose content, was observed by culturing C. disseminatus on a medium with different formulations. Based on the optimal formula, the fermentation conditions were determined by an orthogonal test and EUG was extracted. After that, the characteristic parameters of EUG were determined by Fourier transform infrared (FT-IR), 1hydrogen-nuclear magnetic resonance (1H-NMR), scanning electron microscopy (SEM), polarizing optical microscope (POM), differential scanning calorimeter (DSC), X-ray diffractometer (XRD), and gel permeation chromatography (GPC). The results showed that the lignin and hemicellulose contents in Eucommia leaves decreased significantly with the increase of fermentation days, and C. disseminatus mediated fermentation had an obvious damage effect on the tissue of Eucommia leaves. At the same time, xylan can be used as an initiator to induce the expression of degradation-related enzymes. The extraction yield of EUG from E. ulmoides leaves after fermentation was 3.06%, which was significantly improved. The optimum fermentation conditions were 4% inoculum, 28°C fermentation temperature, and pH = 5. The results of infrared FT-IR and 1H-NMR showed that the extract had typical characteristics of EUG, and the surface and quenching part of the EUG were smooth, as observed by SEM. In addition, DSC and XRD analysis show that there are two crystal types, α and β, in the crystallization process of molten EUG. The main crystallization type of EUG extracted from fermented leaves was β type.
The ability of C. disseminatus to produce degrading enzymes can effectively improve the extraction rate of EUG under optimal fermentation process. The method can maintain the integrity of gutta-percha better. EUG has the characteristics of good crystallinity, many β crystal types and high molecular weight.
确定在毛头鬼伞发酵过程中从杜仲叶中提取杜仲胶(EUG)的最佳条件。同时,对杜仲胶的特性进行表征。
通过在不同配方的培养基上培养毛头鬼伞,观察其分泌降解酶的能力,包括酶活性和木质纤维素含量。基于最佳配方,通过正交试验确定发酵条件并提取杜仲胶。之后,通过傅里叶变换红外光谱(FT-IR)、氢核磁共振(1H-NMR)、扫描电子显微镜(SEM)、偏光光学显微镜(POM)、差示扫描量热仪(DSC)、X射线衍射仪(XRD)和凝胶渗透色谱(GPC)测定杜仲胶的特征参数。结果表明,随着发酵天数的增加,杜仲叶中的木质素和半纤维素含量显著降低,毛头鬼伞介导的发酵对杜仲叶组织有明显的破坏作用。同时,木聚糖可作为诱导剂诱导降解相关酶的表达。发酵后杜仲叶中杜仲胶的提取率为3.06%,显著提高。最佳发酵条件为接种量4%、发酵温度28℃、pH = 5。红外FT-IR和1H-NMR结果表明提取物具有杜仲胶的典型特征,SEM观察到杜仲胶的表面和淬火部分光滑。此外,DSC和XRD分析表明,熔融杜仲胶在结晶过程中有α和β两种晶型。从发酵叶中提取的杜仲胶的主要结晶类型为β型。
在最佳发酵工艺下,毛头鬼伞产生降解酶的能力可有效提高杜仲胶的提取率。该方法能更好地保持杜仲胶的完整性。杜仲胶具有结晶性好、β晶型多、分子量高的特点。