灵芝不同生长期子实体多糖含量及糖代谢相关酶活性研究
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摘要
研究灵芝(Ganoderma lucidum Karst)不同生长阶段子实体多糖含量和糖代谢相关酶活性的变化,初步探讨灵芝子实体多糖含量变化的酶学机制。以袋料栽培灵芝为试材,采用苯酚-硫酸法测定子实体多糖含量,分光光度法测定糖代谢相关酶活性。结果表明,3种糖代谢相关酶(己糖激酶、α-磷酸葡萄糖变位酶、磷酸葡萄糖异构酶)的活性变化趋势与子实体多糖含量的变化趋势基本一致。菌蕾期子实体多糖含量最低,为11.7 mg/g。从菌蕾期开始,随着糖代谢相关酶活性逐渐升高,子实体多糖含量逐渐增加,在子实体成熟后孢子弹射前达到最大值,为23.3 mg/g,此时,己糖激酶、α-磷酸葡萄糖变位酶、磷酸葡萄糖异构酶的活性也达到最大值,分别为23.9、25.1、18.7 U/g。之后子实体多糖含量随着糖代谢相关酶活性的显著下降而降低,孢子弹射后,子实体多糖含量仅为16.3 mg/g。三种糖代谢相关酶活性与灵芝子实体多糖含量呈现正相关,推测己糖激酶、α-磷酸葡萄糖变位酶、磷酸葡萄糖异构酶是灵芝多糖合成的关键酶。
In order to study the enzymatic mechanisms responsible for the change of polysaccharides content, the changes of polysaccharides contents and activities of sugar metabolism-related enzymes from the fruiting bodies of Ganoderma lucidum at different growth stages were investigated. The polysaccharides contents and enzymes activities were determined with phenol-sulfuric acid method and spectrophotometry method, respectively, with bag-cultivated Ganoderma lucidum as test material. The results showed that, the change tendency of activities of three kinds of sugar metabolism-related enzymes(hexokinase, α-phosphoglucomutase and phosphoglucoisomerase) was similar to that of polysaccharides contents. The polysaccharides content was the lowest in the budding stage with the value of 11.7 mg/g.From the budding stage, the polysaccharides contents increased gradually with the increase of the related enzymes activities and reached the maximum value 23.3 mg/g before the spore powder ejection in the maturation stage, while the activities of hexokinase, α-phosphoglucomutase and phosphoglucoisomerase also reached the maximum values(23.9,25.1, 18.7 U/g), respectively. And then the polysaccharides contents decreased with the significant decrease of the related enzymes activities and the polysaccharides content was only 16.3 mg/g after spore ejection. There was a significant positive correlation between the polysaccharides content and the activities of sugar metabolism-related enzymes, which speculated that hexokinase, α-phosphoglucomutase and phosphoglucoisomerase were the key enzymes in the biosynthesis of Ganoderma lucidum fruiting body polysaccharides.
In order to study the enzymatic mechanisms responsible for the change of polysaccharides content, the changes of polysaccharides contents and activities of sugar metabolism-related enzymes from the fruiting bodies of Ganoderma lucidum at different growth stages were investigated. The polysaccharides contents and enzymes activities were determined with phenol-sulfuric acid method and spectrophotometry method, respectively, with bag-cultivated Ganoderma lucidum as test material. The results showed that, the change tendency of activities of three kinds of sugar metabolism-related enzymes(hexokinase, α-phosphoglucomutase and phosphoglucoisomerase) was similar to that of polysaccharides contents. The polysaccharides content was the lowest in the budding stage with the value of 11.7 mg/g.From the budding stage, the polysaccharides contents increased gradually with the increase of the related enzymes activities and reached the maximum value 23.3 mg/g before the spore powder ejection in the maturation stage, while the activities of hexokinase, α-phosphoglucomutase and phosphoglucoisomerase also reached the maximum values(23.9,25.1, 18.7 U/g), respectively. And then the polysaccharides contents decreased with the significant decrease of the related enzymes activities and the polysaccharides content was only 16.3 mg/g after spore ejection. There was a significant positive correlation between the polysaccharides content and the activities of sugar metabolism-related enzymes, which speculated that hexokinase, α-phosphoglucomutase and phosphoglucoisomerase were the key enzymes in the biosynthesis of Ganoderma lucidum fruiting body polysaccharides.
引文
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[9]赵洲,陈向东,王立华,等.灵芝子实体多糖积累和糖代谢相关酶的关系研究[J].中国农学通报,2012,28(22):253-257. DOI:10.3969/j.issn.1000-6850.2012.22.045.
[10]刘高强,赵艳,王晓玲,等.灵芝多糖的生物合成和发酵调控[J].菌物学报,2011,30(2):198-205. DOI:10.13346/j.mycosystema.2011.02.015.
[11] TANG Y J, ZHONG J J. Exopolysaccharide biosynthesis and related enzyme activities of the medicinal fungus, Ganoderma lucidum, grown on lactose in a bioreactor[J]. Biotechnology Letters, 2002, 24(12):1023-1026. DOI:10.1023/A:1015677313598.
[12]胡昭庚.灵芝生产全书[M].北京:中国农业出版社,2004:1-12.
[13]刘建华,张志军.灵芝多糖提取与应用现状[J].保鲜与加工,2003,3(3):9-10. DOI:10.3969/j.issn.1009-6221.2003.03.004.
[2] LIN Z B. Cellular and Molecular Mechanisms of Immunomodulation by Ganoderma lucidum[J]. Journal of Pharmacological Sciences, 2005, 99(2):144-153. DOI:org/10.1254/jphs.CRJ05008X.
[3] RUSSELL R, PATERSON M. Ganoderma-a therapeutic fungal biofactory[J]. Phytochemistry, 2006,67(18):1985-2001. DOI:10.1002/chin.200650268.
[4]韦会平,李学刚,蒲盛才,等.灵芝生长过程中灵芝多糖含量变化规律研究[J].中医药学报,2006,34(5):10-12. DOI:10.3969/j.issn.1002-2392.2006.05.006.
[5]张晓云,李朝谦,杨春清.不同生长期赤灵芝子实体多糖含量变化研究[J].食用菌,2008,30(1):10-11. DOI:10.3969/j.issn.1000-8357.2008.01.006.
[6]林应兴.灵芝子实体不同生长阶段多糖与三萜类物质含量的研究[J].亚热带农业研究,2009,5(3):152-154. DOI:10.3969/j.issn.1673-0925.2009.03.003.
[7]宝萍萍,图力古尔,包海鹰.栽培灵芝不同生长阶段多糖和三萜类化合物的含量测定[J].菌物研究,2011,9(2):110-113,118. DOI:10.3969/j.issn.1672-3538.2011.02.009.
[8]刘健鹏,兰进.栽培灵芝胞外酶活性及多糖变化规律的初步研究[J].中国中药杂志,2009,34(4):483-485. DOI:10.3321/j.issn:1001-5302.2009.04.027.
[9]赵洲,陈向东,王立华,等.灵芝子实体多糖积累和糖代谢相关酶的关系研究[J].中国农学通报,2012,28(22):253-257. DOI:10.3969/j.issn.1000-6850.2012.22.045.
[10]刘高强,赵艳,王晓玲,等.灵芝多糖的生物合成和发酵调控[J].菌物学报,2011,30(2):198-205. DOI:10.13346/j.mycosystema.2011.02.015.
[11] TANG Y J, ZHONG J J. Exopolysaccharide biosynthesis and related enzyme activities of the medicinal fungus, Ganoderma lucidum, grown on lactose in a bioreactor[J]. Biotechnology Letters, 2002, 24(12):1023-1026. DOI:10.1023/A:1015677313598.
[12]胡昭庚.灵芝生产全书[M].北京:中国农业出版社,2004:1-12.
[13]刘建华,张志军.灵芝多糖提取与应用现状[J].保鲜与加工,2003,3(3):9-10. DOI:10.3969/j.issn.1009-6221.2003.03.004.
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