Physiological and Biochemical Properties of Agaricus bisporus in Shaking Flask Culture
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摘要:目的 研究双孢蘑菇W192液体菌种摇瓶培养菌丝体生长规律，为液体种子活力判断提供参考。方法 采用液体摇瓶培养，分别测定了液体培养过程中双孢蘑菇菌丝体生物量、菌丝球数量和直径、发酵液pH值、还原糖和氨基氮含量、羧甲基纤维素酶、淀粉酶、酸性蛋白酶、漆酶胞外酶活性生理生化指标。结果 摇瓶培养第8 d时，双孢蘑菇菌丝体生物量最大，为9.7 mg·mL−1；菌丝球数量最多，为880个·mL−1；菌丝体平均直径最大，为0.809 mm；培养液中还原糖、氨基氮含量最高，分别为5.228、0.079 mg·mL−1；羧甲基纤维素酶、淀粉酶、漆酶酶活性最高，分别为0.69 、2.11 、15.02 U。摇瓶培养第10 d时，酸性蛋白酶活性最高，酶活性为2.93 U。将摇瓶培养8 d的液体种子转接液体培养基中进行模拟发酵罐液体菌种培养，获得的菌丝球数量最多，达115个·mL−1，菌丝体生物量最大，达7.56 mg·mL−1。结论 双孢蘑菇液体菌种活性与上述指标具有一定的相关性，结合液体种子活力验证，判断第8 d的液体菌种活力最高，可作为液体种子用于发酵罐扩繁培养。Abstract:Objective Mycelial growth of Agaricus bisporus W192 in a shaking flask was monitored to determine viability of the culture method for scale-up application.Method Physiological and biochemical properties of A. bisporus mycelia biomass in the liquid culture including the number and diameter of mycelium pellets, pH of fermentation broth, content of reducing sugar and amino nitrogen as well as the activity of extracellular carboxymethyl cellulase, amylase, acid protease, and laccase were monitored for the analysis.Result On the 8th day of culture, the fungal biomass reached a maximum at 9.7 mg·ml−1, the greatest number of mycelium pellets of 880 CFU·mL−1, the largest average diameter of mycelium at 0.809 mm, the peak reducing sugar and amino nitrogen contents at 5.228 and 0.079 mg·ml−1, respectively, and the highest activities of carboxymethyl cellulase, amylase, and laccase of 0.69 , 2.11 , and 15.02 U, respectively. The acid protease activity of 2.93U maxed on the 10th day. By transferring the mycelia in a simulated liquid fermentation tank after 8d of culture, 115 CFU·mL−1 of mycelium pellets and 7.56mg·mL−1 in biomass were obtained.Conclusion The mycelial growth of A. bisporus in the liquid shaking flask culture correlated with some of selected physiological and biochemical indices. The established method was deemed applicable for tank fermentation in propagating A. bisporus for large scale production of the mushroom.
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图 7 不同培养时间胞外酶酶活性的变化
Figure 7. Changes on extracellular enzyme activities in flask culture
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图 8 不同培养时间的液体种子对液体菌种菌丝球数量的影响
Figure 8. Effect of culture time on number of mycelium pellets
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