天麻醇提物对灰树花发酵菌丝体多糖及β-葡聚糖合成量的影响
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摘要
在灰树花液体深层发酵体系中添加天麻醇提物,考察天麻醇提物对灰树花发酵菌丝体生物量、菌丝体多糖及β-葡聚糖含量的影响,并进一步研究起显著促进作用的关键特征成分;同时,采用高效液相色谱法对7 g/L天麻醇提物中的特征成分进行定量分析,研究了对灰树花菌丝体生物量、菌丝体多糖及β-葡聚糖起促进作用的关键特征成分及优化添加量。结果表明,适当浓度的天麻醇提物能够显著提高灰树花菌丝体生物量、菌丝体多糖及β-葡聚糖的含量,在其质量浓度为7g/L时,菌丝体干重、菌丝体多糖及β-葡聚糖含量均达到最大值,分别为1.708 g/L、5. 974%和2. 055 mg/g,与不添加天麻醇提物相比分别增加了0. 36、1. 29和1. 44倍(P<0. 05)。天麻醇提物中的特征成分分别为天麻素5. 837 5 mg/g,对羟基苯甲醇1.107 2 mg/g,对羟基苯甲醛0.660 1 mg/g,对灰树花菌丝体生物量、菌丝体多糖及β-葡聚糖起促进作用的关键特征成分均为对羟基苯甲醛,其.优化添加量分别为100、250、150 mg/L。
Rhizoma gastrodiae ethanol extracts were supplemented into the fermentation system of Grifola frondosa to study effects of R. gastrodiae ethanol extract on mycelia biomass, mycelia polysaccharides andβ-glucan yield of Grifola frondosa by submerged fermentation. In addition,the key ingredients in Rhizoma gastrodiae ethanol extracts were identified. Meanwhile, the appropriate concentration of characteristic constituents of R. gastrodiae ethanol extracts were determined by high performance liquid chromatography. The results showed that R. gastrodiae ethanol extracts with the concentration of 7 g/L could significantly promote cell growth, mycelia polysaccharides, and β-glucan yield. Under this concentration, the biomass, mycelia polysaccharides and β-glucan yield reached 1. 708 g/L, 5. 974% and 2. 055 mg/g, and significantly(P < 0. 05) increased by 0.36, 1.29, and 1.44 times, respectively, compared with the control( without additives). The characteristic ingredients of R. gastrodiae ethanol extracts were gastrodin(GA), p-hydroxybenzyl alcohol( HA),and p-hydroxylbenzaldehyde( HBA),and the contents were5. 837 5,1. 107 2,and 0. 660 1 mg/g,respectively. Meanwhile, HBA was the most responsible ingredients to promote the biomass, mycelia polysaccharides and β-glucan yield in submerged culture of Grifola frondosa and the optimum supplemental levels were 100, 250, and 150 mg/L, respectively.
Rhizoma gastrodiae ethanol extracts were supplemented into the fermentation system of Grifola frondosa to study effects of R. gastrodiae ethanol extract on mycelia biomass, mycelia polysaccharides andβ-glucan yield of Grifola frondosa by submerged fermentation. In addition,the key ingredients in Rhizoma gastrodiae ethanol extracts were identified. Meanwhile, the appropriate concentration of characteristic constituents of R. gastrodiae ethanol extracts were determined by high performance liquid chromatography. The results showed that R. gastrodiae ethanol extracts with the concentration of 7 g/L could significantly promote cell growth, mycelia polysaccharides, and β-glucan yield. Under this concentration, the biomass, mycelia polysaccharides and β-glucan yield reached 1. 708 g/L, 5. 974% and 2. 055 mg/g, and significantly(P < 0. 05) increased by 0.36, 1.29, and 1.44 times, respectively, compared with the control( without additives). The characteristic ingredients of R. gastrodiae ethanol extracts were gastrodin(GA), p-hydroxybenzyl alcohol( HA),and p-hydroxylbenzaldehyde( HBA),and the contents were5. 837 5,1. 107 2,and 0. 660 1 mg/g,respectively. Meanwhile, HBA was the most responsible ingredients to promote the biomass, mycelia polysaccharides and β-glucan yield in submerged culture of Grifola frondosa and the optimum supplemental levels were 100, 250, and 150 mg/L, respectively.
引文
[1]聂文强,吴天祥,钟敏,等.真菌灰树花菌丝体转录组测序及分析[J].食品科学,2017,38(20):13-18.NIE W Q, WU T X, ZHONG M, et al. Transcriptome sequencing and analysis of Grifola frondosa mycelia[J].Food Science, 2017,38(20):13-18.
[2]徐晓宝,吴天祥.灰树花发酵过程天麻成分变化的HPLC检测方法研究[J].中国酿造,2012, 31(5):182-185.XU X B, WU T X. Determination of extracts from Gastrodia tuber fermented with Gifola frondosa by HPLC method[J].China Brewing,2012, 31(5):182-185.
[3] MCLNT,SH M, STONE B A, STANISICH V A. Curdlan and other bacterial(1→3)-β-D-glucans[J]. Applied Microbiology and Biotechnology, 2005, 68(2):163-173.
[4] FANG J, WANG Y, LV X, et al. Structure of aβ-glucan from Grifola frondosa and its antitumor effect by activating dectin-1/Syk/NF-κB signaling[J]. Glycoconjugate Journal, 2012, 29(5/6):365-377.
[5] HONG J FH, JUNG H K. Antioxidant and antitumor activities ofβ-ghucan-rich exopolysaccharides with different molecular weight from Paenibacillus polymyxa JB115[J].Journal of Korean Society for Applied Biological Chemistry,2014, 57(1):105-112.
[6] SONG G, DU Q. Structure characterization and antitumor activity of anαβ-glucan polysaccharide from Auricularia polytricha[J]. Food Research International, 2012, 45(1):381-387.
[7]吴昊,张建法.β-葡聚糖免疫调节作用的研究进展[J].细胞与分子免疫学杂志,2014, 30(1):97-100.WU H, ZHANG J F. Research progress about immunomodulatory effect ofβ-glucan glucan[J]. Chinese Journal of C.ellular and Molecular Immunology, 2014, 30(1):97-100.
[8] SMIDERLE F R, BAGGIO C H, BORATO D G, et al.Anti-inflammatory properties of the medical mushroom Cordyceps militaris might be related to its linear(1→3)-β-D-glucan[J]. PLOS One, 2014, 9(10):1-11.
[9] JUNG K,HA Y, HA S K, et al. Antiviral effect of Soccharormyces cerevisiae beta-glucan to swine influenza virus by increased production of interferon-gamma and nitric oxide[J]. Zoonoses&Public Health, 2010, 51(2):72-76.
[10] MAITY P, SEN I K, MAJN P K, et al. Structural, immunological,and antioxidant studies ofβ-glucan fromedible mushroom Eutoloma lividoalbum[J]. Carbohydrate Polymers, 2015, 123:350-358.
[11] KAO P F, WANG S H, HUNG W T,et al. Structural characterization and antioxidative activity of low-molecularweights beta-1, 3-glucan from the residue of extracted Ganoderma lucidum fruiting bodies[J]. Journal of Biomedicine and Biotechnology, 2015,2012(1):673-764.
[12] MARTINICHENHERRERO J C, CARBONERO E R,PAJ G, et al. Anticoagulant and antithrombotic activity of a sulfate obtained from a glucan component of the lichen Parmotrema mantiqueirense Hale[J]. Carbohydrate Polymers, 2005, 60(1):7-13.
[13] UAN X H, LI Z L, YANG D S, et al. Study on the chemical constituents of Gastrodia elata[J]. Journal of Chinese Medicinal Materials, 2013, 36(10):1608-1611.
[14]王亚威,李志峰,何明珍,等.天麻化学成分研究[J].中草药,2013, 44(21):2974-2976.WANG Y W, LI Z F, HE M Z, et al. Chemical constituents of Gastrodia elata[J]. Chinese Traditional and Herbal Drugs, 2013, 44(21):2974-2976.
[15]段小花,李资磊,杨大松,等.昭通产天麻化学成分研究[J].中药材,2013, 36(10):1608-1611.DUAN X H, LI Z L, YANG D S, et al. Study on the chemical constituents of Gastrodia elata[J]. Journal of C.hinese Medicinal Materials, 2013, 36(10):1608-1611.
[16] YANG H L, WU T X, ZHANG K C. Enhancement of mycelial growth and polysaccharide production in Ganoderma lucidum(the Chinese medicinal fungus,'Lingzhi')by the addition of ethanol[J]. Biotechnology Letters, 2004, 26(10):841-844.
[17] 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.
[18]张勇,吴天祥,徐晓宝,等.天麻提取物的制备及其对灰树花发酵的影响[J].食品与机械,2012, 28(1):150-153.ZHANG Y, WU T X, XU X B, et al. Preparation of Gastrodia elata BL. extracts and their effects on Grifolafrondosa in submerged culture[J]. Food&Machinery,2012, 28(1):150-153.
[19] ZHANG Y, WANG N, WU T X. Effect of the extracts from Gastrodia elata BL. on mycelial growth and polysaccharide biosynthesis by Grifola frondosa[J]. African Journal of Microbiology Research, 2012, 6(2):379-384.
[20] XU X B, WU T X, WANG F. The effect of exopolysaccharide biosynthesis and related enzyme activities of Grifola frondosa by the addition of ethanol extracts from traditional Chinese medicine, Gastrodia tuber[J]. African Journal of Biotechnology, 2012, 11(15):3656-3662.
[21] WANG N, WU T X, ZHANG Y, et al. Experimental analysis on the effect of addition of Rhizoma gastrodiae on mycelia and exopolysaccharide productions by submerged culture of Grifola frondosa[J]. African Journal of Biotechnology, 2012, 11(20):4666-4672.
[22]贺宗毅,吴天祥,徐晓宝.中药天麻成分对灰树花胞外多糖合成及相关关键酶的影响[J].食品科学,2013, 34(11):199-202.HE Z Y,WU T X, XU X B. Effect of Rhizoma gastrodiae on key enenzyme activities involved in the biosynthesis of exopolysaccharides from Grifola frondosa in submerged culture[J]. Food Science, 2013, 34(11):199-202.
[23] WU C Y, WU T X. Effect of the main ingredients of Rhizoma gastrodiae on mycelial biomass and exopolysaccharide productions by submerged culture of Grifola frondosa[J]. International Journal of Food Science and Technology, 2015, 50:1726-1730.
[24] DUBOIS M, GILLES K A, HAMILTON J K,et al.Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry 1956,28:350-356.
[25]熊婷.灵芝发酵物中功能性β-葡聚糖的提取技术研究[D].株洲:中南林业科技大学,2015.
[26]上官端琳,吴素蕊,赵天瑞,等.α-萘乙酸对黑脉羊肚菌生物量、胞内多糖含量及多糖抗氧化性能的影响[J].食品与发酵工业,2013, 39(11):159-162.
[27]朱俊杰.天麻醇提物对灰树花深层发酵代谢影响及其机制研究[D].贵阳:贵州大学,2016.
[2]徐晓宝,吴天祥.灰树花发酵过程天麻成分变化的HPLC检测方法研究[J].中国酿造,2012, 31(5):182-185.XU X B, WU T X. Determination of extracts from Gastrodia tuber fermented with Gifola frondosa by HPLC method[J].China Brewing,2012, 31(5):182-185.
[3] MCLNT,SH M, STONE B A, STANISICH V A. Curdlan and other bacterial(1→3)-β-D-glucans[J]. Applied Microbiology and Biotechnology, 2005, 68(2):163-173.
[4] FANG J, WANG Y, LV X, et al. Structure of aβ-glucan from Grifola frondosa and its antitumor effect by activating dectin-1/Syk/NF-κB signaling[J]. Glycoconjugate Journal, 2012, 29(5/6):365-377.
[5] HONG J FH, JUNG H K. Antioxidant and antitumor activities ofβ-ghucan-rich exopolysaccharides with different molecular weight from Paenibacillus polymyxa JB115[J].Journal of Korean Society for Applied Biological Chemistry,2014, 57(1):105-112.
[6] SONG G, DU Q. Structure characterization and antitumor activity of anαβ-glucan polysaccharide from Auricularia polytricha[J]. Food Research International, 2012, 45(1):381-387.
[7]吴昊,张建法.β-葡聚糖免疫调节作用的研究进展[J].细胞与分子免疫学杂志,2014, 30(1):97-100.WU H, ZHANG J F. Research progress about immunomodulatory effect ofβ-glucan glucan[J]. Chinese Journal of C.ellular and Molecular Immunology, 2014, 30(1):97-100.
[8] SMIDERLE F R, BAGGIO C H, BORATO D G, et al.Anti-inflammatory properties of the medical mushroom Cordyceps militaris might be related to its linear(1→3)-β-D-glucan[J]. PLOS One, 2014, 9(10):1-11.
[9] JUNG K,HA Y, HA S K, et al. Antiviral effect of Soccharormyces cerevisiae beta-glucan to swine influenza virus by increased production of interferon-gamma and nitric oxide[J]. Zoonoses&Public Health, 2010, 51(2):72-76.
[10] MAITY P, SEN I K, MAJN P K, et al. Structural, immunological,and antioxidant studies ofβ-glucan fromedible mushroom Eutoloma lividoalbum[J]. Carbohydrate Polymers, 2015, 123:350-358.
[11] KAO P F, WANG S H, HUNG W T,et al. Structural characterization and antioxidative activity of low-molecularweights beta-1, 3-glucan from the residue of extracted Ganoderma lucidum fruiting bodies[J]. Journal of Biomedicine and Biotechnology, 2015,2012(1):673-764.
[12] MARTINICHENHERRERO J C, CARBONERO E R,PAJ G, et al. Anticoagulant and antithrombotic activity of a sulfate obtained from a glucan component of the lichen Parmotrema mantiqueirense Hale[J]. Carbohydrate Polymers, 2005, 60(1):7-13.
[13] UAN X H, LI Z L, YANG D S, et al. Study on the chemical constituents of Gastrodia elata[J]. Journal of Chinese Medicinal Materials, 2013, 36(10):1608-1611.
[14]王亚威,李志峰,何明珍,等.天麻化学成分研究[J].中草药,2013, 44(21):2974-2976.WANG Y W, LI Z F, HE M Z, et al. Chemical constituents of Gastrodia elata[J]. Chinese Traditional and Herbal Drugs, 2013, 44(21):2974-2976.
[15]段小花,李资磊,杨大松,等.昭通产天麻化学成分研究[J].中药材,2013, 36(10):1608-1611.DUAN X H, LI Z L, YANG D S, et al. Study on the chemical constituents of Gastrodia elata[J]. Journal of C.hinese Medicinal Materials, 2013, 36(10):1608-1611.
[16] YANG H L, WU T X, ZHANG K C. Enhancement of mycelial growth and polysaccharide production in Ganoderma lucidum(the Chinese medicinal fungus,'Lingzhi')by the addition of ethanol[J]. Biotechnology Letters, 2004, 26(10):841-844.
[17] 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.
[18]张勇,吴天祥,徐晓宝,等.天麻提取物的制备及其对灰树花发酵的影响[J].食品与机械,2012, 28(1):150-153.ZHANG Y, WU T X, XU X B, et al. Preparation of Gastrodia elata BL. extracts and their effects on Grifolafrondosa in submerged culture[J]. Food&Machinery,2012, 28(1):150-153.
[19] ZHANG Y, WANG N, WU T X. Effect of the extracts from Gastrodia elata BL. on mycelial growth and polysaccharide biosynthesis by Grifola frondosa[J]. African Journal of Microbiology Research, 2012, 6(2):379-384.
[20] XU X B, WU T X, WANG F. The effect of exopolysaccharide biosynthesis and related enzyme activities of Grifola frondosa by the addition of ethanol extracts from traditional Chinese medicine, Gastrodia tuber[J]. African Journal of Biotechnology, 2012, 11(15):3656-3662.
[21] WANG N, WU T X, ZHANG Y, et al. Experimental analysis on the effect of addition of Rhizoma gastrodiae on mycelia and exopolysaccharide productions by submerged culture of Grifola frondosa[J]. African Journal of Biotechnology, 2012, 11(20):4666-4672.
[22]贺宗毅,吴天祥,徐晓宝.中药天麻成分对灰树花胞外多糖合成及相关关键酶的影响[J].食品科学,2013, 34(11):199-202.HE Z Y,WU T X, XU X B. Effect of Rhizoma gastrodiae on key enenzyme activities involved in the biosynthesis of exopolysaccharides from Grifola frondosa in submerged culture[J]. Food Science, 2013, 34(11):199-202.
[23] WU C Y, WU T X. Effect of the main ingredients of Rhizoma gastrodiae on mycelial biomass and exopolysaccharide productions by submerged culture of Grifola frondosa[J]. International Journal of Food Science and Technology, 2015, 50:1726-1730.
[24] DUBOIS M, GILLES K A, HAMILTON J K,et al.Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry 1956,28:350-356.
[25]熊婷.灵芝发酵物中功能性β-葡聚糖的提取技术研究[D].株洲:中南林业科技大学,2015.
[26]上官端琳,吴素蕊,赵天瑞,等.α-萘乙酸对黑脉羊肚菌生物量、胞内多糖含量及多糖抗氧化性能的影响[J].食品与发酵工业,2013, 39(11):159-162.
[27]朱俊杰.天麻醇提物对灰树花深层发酵代谢影响及其机制研究[D].贵阳:贵州大学,2016.