灵芝真菌液体发酵及其产物应用的研究进展

灵芝作为一种白腐真菌,同时也是珍稀的食药用真菌,富含多种生物活性成分。液体发酵技术生产周期短、效率高、产量大、品质稳定,是开发利用灵芝资源的重要途径。近年来,灵芝属真菌菌丝体液体发酵技术的开发与应用取得了较大进展。本文对灵芝属真菌液体发酵产物的主要活性成分及其药用效果、液体发酵工艺优化和发酵产物的应用进行综述,并对本领域的未来进行展望。     

灵芝液体发酵条件的研究

本文研究了某些因素对灵芝液体发酵的影响。4天菌龄的液体菌种发酵效果最好;300毫升三角瓶以装入60毫升培养液发酵较合适;蔗糖、花生饼粉分别为灵芝液体发酵的理想碳、氮源;液体发酵培养基最佳配方为:蔗糖4％、花生饼粉3％、硫酸铵0.15％、磷酸二氢钾0.15％,可得灵芝菌丝体百分干重为0.83,每100毫升发酵液得粗多糖0.3克。     

灵芝MP-01菌株液体发酵培养基的筛选

对灵芝MP-01菌株在三种培养基上的生长性状进行了研究,并对其液体发酵过程中的pH值变化、发酵全液的多糖含量变化、菌丝生物量及发酵培养性状等进行了观测。结果表明,玉米面培养基是灵芝MP-01菌株液体发酵的适宜培养基,其适宜的终止发酵时间在培养的第6 d,此时发酵液多糖含量达到10.32 mg/mL。     

虎乳灵芝液体发酵培养研究

目的:建立虎乳灵芝菌丝液体发酵培养方法。方法:采用改变培养基的pH和选择不同的碳源及氮源,通过比较虎乳灵芝菌丝的低温冷冻干燥重量,分别研究菌丝生长的最适培养基。结果:虎乳灵芝菌丝最适生长的液体培养基pH 6~7(中性微偏弱酸);菌丝生长的最好的液体培养基中的碳源为葡萄糖,氮源为酵母粉。结论:葡萄糖 酵母粉液体培养基可以作为虎乳灵芝菌丝大量发酵培养的培养基。     

玉米水解糖液体培养灵芝发酵条件的优化

目的：优化液体培养灵芝的发酵条件,提高多糖产量。方法：采用玉米水解糖为主要成分的培养基,通过单因素和正交实验,对赤芝G22菌株液体培养过程中影响多糖产量的发酵温度、摇床转速等工艺条件进行了研究。结果：经极差分析和方差分析确定了多糖高产的最佳发酵条件为：发酵温度27℃、摇床转速170r/min、培养基初始pH值6.5、发酵时间144 h。结论：通过优化液体发酵条件,可显著提高灵芝多糖的产量。在最佳发酵条件下液体培养G22菌株,灵芝总多糖产量由1.851g/L提高到2.439g/L,提高了31.0%。     

灵芝-银杏双向液体发酵条件优化及抗氧化的研究

本研究采用双向液体发酵技术,以灵芝作为出发菌株,银杏叶为药性基质,以发酵产物的抗氧化活性为检测指标,通过单因素及正交实验,对双向发酵条件进行了优化,并进一步对发酵后的活性物质进行了检测。结果表明,灵芝‐银杏双向液体发酵的最佳条件为:银杏叶接种量为0.15g/100m L、初始p H 8.0、发酵时间9d、添加银杏叶的发酵时间为第2天,在此基础上,菌株的还原力达到0.731;活性物质多糖含量为245.20mg/g,三萜含量为70.96mg/g,总黄酮质含量为10.98mg/g,分别是对照的2.38倍、1.96倍和2.10倍。     

灵芝多糖液体发酵条件的研究

液体发酵灵芝是目前灵芝多糖开发的有效途径。以灵芝的生物量和胞外多糖为指标,对影响灵芝发酵的条件进行了研究。单因素实验表明灵芝发酵的最佳碳源、(?)源和生长因子分别是葡萄糖、酵母膏和维生素B1,最适温度、起始pH值和摇床转速分别是28℃、5．5和160 rpm。最佳培养方式是接种后静置4 h再振荡培养,其生物量和胞外多糖的产量最高,分别为7．743g/L和0．907g/L。     

灵芝液体发酵清除自由基活性产物发酵条件的优化

以红灵芝(Ganoderma.lucidum)为实验菌株,对其液体发酵活性产物清除自由基的发酵条件进行了研究。实验考察了培养基成分对灵芝液体发酵所产生的有效产物清除自由基能力的影响。结果表明,葡萄糖、酵母粉分别为灵芝发酵活性产物清除自由基效果较合适的碳源、氮源,最佳质量浓度分别为40 g.L-1和3 g.L-1,清除自由基的能力分别为55.7%、40.8%和66.5%、50.6%;铁离子对灵芝发酵产物清除超氧阴离子具有明显的效果,适宜质量分数为70×10-6;清除超氧阴离子的能力为54.6%,而硒离子对灵芝发酵产物抑制羟自由基有明显的促进作用,最适的质量分数为90×10-6清除羟自由基的能力为67.7%。油酸能促进灵芝发酵产物的清除超氧阴离子和羟自由基的能力,适宜的质量分数为0.1%,清除两种自由基的能力分别为:73.7%,53%。     

13种野生灵芝菌丝体中胞内三萜与多糖含量的比较

为探究不同野生灵芝的主要活性成分以及对野生灵芝的开发利用价值,对13种野生灵芝菌株在同一条件下进行液体发酵,采用化学分析的方法,比较菌丝体胞内三萜和多糖的含量差异。结果显示,13种灵芝菌株的三萜和多糖含量有很大差异,其中无柄紫灵芝Ganoderma mastoporum、亮盖灵芝G. lucidum和树舌灵芝G. applanatum的三萜含量较高;树舌灵芝G. applanatum、紫芝G. sinense和褐灵芝G. brownii的多糖含量较高。目前国内广泛栽培灵芝G. lingzhi的野生菌株发酵产物中的三萜和多糖含量并不是最高的,研究结果表明不同种类的野生灵芝还有进一步挖掘的潜在价值。     

甘蔗纤维固定灵芝菌丝及其在液体发酵中的应用

以甘蔗纤维作为灵芝菌丝固定载体,通过扫描电子显微镜观察确定载体固定时间,通过液体发酵灵芝菌丝球大小形态、生物量、胞外多糖和胞外三萜含量确定载体形状、大小与数量。结果表明,灵芝菌丝在载体上固定时间为7d,载体为1.5cm×1.5cm（直径×高度）的圆柱体小块（Y1.5）,接种数量6块,可连续稳定发酵7代。以甘蔗纤维固定发酵制备灵芝液体发酵种子,发酵后菌丝球大小均一,生物量、胞外多糖含量和三萜含量分别提高78%、84%和60%。     

An unstructured kinetic model for the improvement of triterpenes production by Ganoderma lucidum G0119 based on nitrogen source effect

The kinetics of cell growth and triterpenes production for liquid submerged fermentation of the medicinal mushroom Ganoderma lucidum were investigated. A kinetic model was developed based on the Logistic and Luedeking-Piret equations for cell growth, substrate consumption and triterpene formation. The kinetic parameters of the model were optimized by specifically designed Runge-Kutta genetic algorithms. The mathematical model simulated the experimental data well and was capable of explaining the behavior of triterpenes production. The predictions of the kinetics from this model are very good both for normal fermentation kinetics under nitrogen limitation as well as for predictions of transitions to sluggish fermentations. The resulting model is very useful for scaling up liquid submerged fermentation of the mushroom G. lucidum and its application to the industrial production of triterpene.     

影响灵芝三萜类物质液态发酵合成的关键因素研究进展

灵芝是我国名贵的食药两用型菌类,具有广泛的药用价值,其三萜类物质为灵芝中最主要的药理活性物质之一。灵芝液态发酵因具有生长周期短、环境条件可控、目标产物质量稳定及易实现规模化制备等特点而成为获得灵芝三萜类物质最有前景的方式。灵芝三萜代谢途径、发酵工艺及参数、溶解氧控制等是影响灵芝三萜类物质液态发酵合成的关键因素。本文总结了灵芝三萜生物合成的代谢途径和相关的酶(基因)、液态发酵方式和发酵参数调节的溶解氧控制这3个层面对灵芝三萜类物质生物合成的影响,并对今后的研究方向进行了展望,为液态培养灵芝三萜类物质调控及高产提供参考,也为下一步研究提供借鉴。     

灵芝液态深层发酵三萜类化合物研究进展

三萜类化合物是灵芝中主要的活性化学成分之一,由于其具有多种重要的生理活性,现已成为国内外学者研究灵芝的热点。本文总结了灵芝三萜发酵工艺的优化及其生物合成中的信号转导等方面的进展,并在此基础上提出了灵芝发酵研究中存在的问题,以期为灵芝三萜液态深层发酵的调控研究及发酵生产工艺的开发提供参考和启示。     

樟芝产生三萜类化合物的液体培养条件的响应面法优化

樟芝(Antrodia cinnamomea)是一种非褶菌目、多孔菌科的多年生蕈菌类,樟芝子实体中含有多种生理活性物质包括三萜类化合物,但野生樟芝只生长在台湾特有的牛樟树树干腐朽的心材内壁和枯死倒伏的牛樟树表面,生长极其缓慢,很难获得,价格昂贵。本研究的目的是通过液体培养樟芝的菌丝体以产生三萜类化合物,用响应面法优化了樟芝产生三萜类化合物的液体发酵条件。首先对樟芝的液体发酵的培养基成分及培养条件进行了单因素优化,研究发现樟芝液体发酵产生三萜类化合物的最佳碳源和氮源分别为葡萄糖和酵母提取物,最佳浓度分别为100 g/L和9 g/L,优化的培养基配方为(g/L):葡萄糖100.0,酵母提取物9.0,Mg SO4·7H2O 0.5,KH2PO41.0。优化的培养条件为:初始pH值6.0,温度28℃,接种量10%(v/v),摇床转速180 r/min。在此基础上,用响应面法对樟芝液体发酵产生三萜类化合物影响最大的三个因素即碳源、氮源和培养温度进行了优化,结果优化的三种因素条件为:葡萄糖104.71 g/L,酵母提取物9.93 g/L,温度28.42℃。用优化的培养条件液体培养樟芝,其三萜类化合物的产量达到391.45 mg/L,比优化前的产量223.39 mg/L提高了75.23%,为进一步研究液体培养樟芝产生三萜类化合物奠定了基础。     

雷公藤解毒持效双向发酵工艺的建立

采用双向发酵的原理,运用"发酵过程动态比较法"研究了灵芝接种于雷公藤药性基质上发酵不同时间所得菌质的化学成分、急性毒性和免疫功能的变化,以其确定雷公藤解毒持效双向发酵的最佳发酵终点。研究结果表明:发酵第30天所得的菌质(G30)总二萜的含量最低,为0.57%;与雷公藤生药比较,G30的LD50最高,且发酵第30天所得菌质的体液免疫和细胞免疫抑制作用最强。综合对成分含量、毒性及药效的动态数据联系比较、分析,确定了雷公藤解毒持效双向发酵的发酵终点与适宜的发酵周期是菌丝长满瓶后的第30天。     

Treatment of coloured industrial effluents with Pleurotus spp

The decolouration of fermentation residues (vinasse) and liquid extract of coffee pulp by the mushroom Pleurotus ostreatus was studied in addition to laccase activity. The fungus was inhibited in both residues when they remained undiluted. In submerged cultivation on wastewaters a good production of biomass (14.8 g/l for vinasse and 5.4 g/l for extract of coffee pulp) and also laccase activity (14.1 U/ml for vinasse and 3.0 U/ml for extract of coffee pulp) up to the 10 days of fermentation was observed, being significantly greater in the culture with vinasse. It was shown that treatment with this mushroom reduces both the chemical oxygen demand and the colour, contributing to their biological treatment.     

Effects of fermentation products of Ganoderma lucidum on growth performance and immunocompetence in weanling pigs

The purpose of this study was to test fermentation, for its products of a Chinese medicinal mushroom, Ganoderma lucidum, cultured by submerged fermentation for its effect on growth performance and immunocompetence in weanling piglets. In Experiment 1, 72 weanling piglets were allotted to one of four treatments receiving these fermentation products (GLF, expressed as amount of beta-glucans) at 0 (control), 50, 100, and 150 mg/kg feed for 4 weeks. The results showed that at a supplementation level of 50 mg/kg feed, GLF caused the best growth performance, the highest pseudorabies antibody titre, and a decrease of blood glucose level. It was also demonstrated that GLF up-regulated the cell-mediated immune response related cytokines (IL-2, IFN-gamma, and TNF-alpha) expression in different lymphoid tissues. After challenging with porcine circovirus (PCV) type 2 (Experiment 2), a supplementation with 50 mg GLF per kg feed also inhibited PCV-2 virus amplification, and ameliorated lymphocyte depletion in different lymphoid tissues. Conclusively, feed supplemented with GLF at 50 mg/kg could be beneficial to counteract the physiological stress in weanling piglets.     

Effects of fermentation products of Ganoderma lucidum on growth performance and immunocompetence in weanling pigs

The purpose of this study was to test fermentation, for its products of a Chinese medicinal mushroom, Ganoderma lucidum, cultured by submerged fermentation for its effect on growth performance and immunocompetence in weanling piglets. In Experiment 1, 72 weanling piglets were allotted to one of four treatments receiving these fermentation products (GLF, expressed as amount of β-glucans) at 0 (control), 50, 100, and 150 mg/kg feed for 4 weeks. The results showed that at a supplementation level of 50 mg/kg feed, GLF caused the best growth performance, the highest pseudorabies antibody titre, and a decrease of blood glucose level. It was also demonstrated that GLF up-regulated the cell-mediated immune response related cytokines (IL-2, IFN-γ, and TNF-α) expression in different lymphoid tissues. After challenging with porcine circovirus (PCV) type 2 (Experiment 2), a supplementation with 50 mg GLF per kg feed also inhibited PCV-2 virus amplification, and ameliorated lymphocyte depletion in different lymphoid tissues. Conclusively, feed supplemented with GLF at 50 mg/kg could be beneficial to counteract the physiological stress in weanling piglets.     

Optimization of nutrient medium for submerged cultivation of Ganoderma lucidum (Curt.: Fr.) P. Karst

The dependence of the amount of the grown vegetative mycelium of Ganoderma lucidum on the composition of the nutrient medium has been studied under conditions of submerged cultivation. The medium was optimized using full factorial and steepest ascent experimental designs. The addition of two carbon sources to the medium considerably improved the submerged growth of the mushroom. An optimized medium provided for a high yield (20-20.95 g/l) of the morphologically homogeneous mycelium and shortened the cultivation period to 3–4 days.     

Solid-state (solid-substrate) food/beverage fermentations involving fungi

“Solid-substrate” fermentation developed in the Orient is a very useful fermentation method. It is presently used to produce a variety of foods, beverages and related products. Solid-substrate fermentation products utilizing fungi including soy sauce, miso and tempe, ontjom, sake, and bread have been produced for centuries at the home and village level. They are examples of economical methods of preserving and improving the flavor, texture and nutritive values of cereal/legume substrates. “Solid-substrate” fermentation is also applied to animal products such as milk to produce Roquefort and Camembert cheeses which diversify the food flavors available to man “Solid-substrate”fermentation has certain advantages. The substrate is concentrated; the product can be extracted with relatively small quantities of solvent; the product can be easily dehydrated; moisture level can be controlled favoring the desired organisms; enzyme concentration is generally higher than is submerged fermentation; product concentration is generally higher than in submerged cultures; it is the only technique that yields true mushroom fruiting bodies and it can be used not only for production of crude enzyme concentrates (koji) but also for raising the protein content of high starch substrates. It also can be used to increase the content of vitamins at low cost. Disadvantages of “solid-substrates”from the modern industrial processing view point are the greater difficulty of handling solid substrate and the greater difficulty of controlling the fermentation parameters, temperature, pH and oxygen, and rate of microbial growth compared with liquid submerged fermentations.