利用扁桃斑鸠菊叶发酵生产蛹虫草胞外多糖的条件及其抗氧化活性
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摘要
蛹虫草胞外多糖具有增强免疫力、抗疲劳等药理活性,有极高的保健价值。为高效地获取蛹虫草胞外多糖,本研究通过向发酵培养基中添加适量的扁桃斑鸠菊叶粉末,来提高蛹虫草发酵液中胞外多糖的产量,并对优化得到的胞外多糖红外吸收光谱和化学抗氧化活性进行了研究。实验结果表明,液体发酵最优条件为:扁桃斑鸠菊叶粉末添加量8 g/L、发酵时间9 d、pH 6.5、接种量5.0 mL,在此条件下,蛹虫草胞外多糖的产量可达(5.24±0.28) mg/mL,与未添加扁桃斑鸠菊叶的空白组相比,胞外多糖产量提高了约205.20%;红外分析与抗氧化活性实验结果显示,扁桃斑鸠菊叶对蛹虫草生产的胞外多糖结构和活性影响较小。该研究结果表明扁桃斑鸠菊叶能够有效地提高蛹虫草胞外多糖的产量,为蛹虫草胞外多糖的高效生产提供了新思路。
Cordyceps militaris exopolysaccharides(EPS) have many pharmacological activities such as boosting immunity and antifatigue. To obtain EPS efficiently, we added moderate Vernonia amygdalina leaf powder as inducer to the fermentation medium to promote the production of Cordyceps militaris EPS and studied the infrared absorption spectrum and antioxidant activities of the EPS after optimization. The optimum liquid fermentation conditions were as follows: addition of Vernonia amygdalina leaf powder of 8 g/L, fermentation duration of 9 d, initial pH of 6.5, inoculation quantity of 5.0 mL.Under such a condition, the yield of Cordyceps militaris EPS reached(5.24±0.28) mg/mL, increased by 205.20% compared to the control group without adding Vernonia amygdalina leaf powder. Results of infrared analysis and antioxidant activity showed that the Vernonia amygdalina leaves had little effect on the structure and activities of Cordyceps militaris EPS. The results of this research suggest that Vernonia amygdalina leaf can enhance the production of Cordyceps militaris EPS effectively, and provides a novel method for efficient production of EPS in Cordyceps militaris.
Cordyceps militaris exopolysaccharides(EPS) have many pharmacological activities such as boosting immunity and antifatigue. To obtain EPS efficiently, we added moderate Vernonia amygdalina leaf powder as inducer to the fermentation medium to promote the production of Cordyceps militaris EPS and studied the infrared absorption spectrum and antioxidant activities of the EPS after optimization. The optimum liquid fermentation conditions were as follows: addition of Vernonia amygdalina leaf powder of 8 g/L, fermentation duration of 9 d, initial pH of 6.5, inoculation quantity of 5.0 mL.Under such a condition, the yield of Cordyceps militaris EPS reached(5.24±0.28) mg/mL, increased by 205.20% compared to the control group without adding Vernonia amygdalina leaf powder. Results of infrared analysis and antioxidant activity showed that the Vernonia amygdalina leaves had little effect on the structure and activities of Cordyceps militaris EPS. The results of this research suggest that Vernonia amygdalina leaf can enhance the production of Cordyceps militaris EPS effectively, and provides a novel method for efficient production of EPS in Cordyceps militaris.
引文
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[9]Ye WJ.Studies on optimization of fermentation conditions,structure and antioxidant activity of Cordyceps militaris polysaccharides[D].Wuhan:Huazhong Agricultural University,2014(in Chinese).叶文姣.蛹虫草多糖的发酵条件优化、结构及抗氧化活性研究[D].武汉:华中农业大学,2014.
[10]Wang YM,Zhu WX,An LG,et al.Optimized cultivation conditions and production efficiency of mycelium and lixiviated cordycepic polysaccharide of Cordyceps militaris.J Unive Jinan:Sci&Tech2010,24(2):148-151(in Chinese).王永敏,祝文兴,安利国,等.优化条件对蛹虫草菌丝体与胞外多糖得率研究.济南大学学报:自然科学版,2010,24(2):148-151.
[11]Yang Z.Antitumor research and clinical applications of bitter leaf.J Nanjing Univ Tradit Chin Med,2013,29(4):397-400(in Chinese).杨早.南非叶化学成分及药理作用研究进展.南京中医药大学学报,2013,29(4):397-400.
[12]Huang W,Li XQ,Liu J,et al.Effects of different hormone combinations on callus inducing of shoot explant from bitter leaf(Vernonia amygdallina Del).J Trop Biol,2017,8(4):419-423(in Chinese).黄伟,黎秀琼,刘娇,等.不同激素配比对南非叶茎愈伤组织诱导的影响.热带生物学报,20178(4):419-423.
[13]Audu SA,Taiwo AE,Ojuolape AR.A study review of documented phytochemistry of Vernonia amygdalina(family asteraceae)as the basis for pharmacologic activity of plant extract.J Natl Sci Res,2012,2(7):1-8.
[14]Chen CF,Xiang XY,Gu Q,et al.Submerged fermentation conditions for synthesis of extracellular polysaccharide by Inonotus obliquus.Chin Tradit Herbal Drugs,2007,38(3):358-361(in Chinese).陈才法,项小燕,顾琪,等.桦褐孔菌胞外多糖合成的深层发酵条件研究.中草药,2007,38(3):358-361.
[15]Meng FL,Song ZF,Liu XX,et al.Determination of cordycepin and total sugar in Cordyceps sinensis and Cordyceps militaris.Food Res Dev,2017,38(21):146-149(in Chinese).孟繁磊,宋志峰,刘笑笑,等.冬虫夏草与蛹虫草中虫草素和总糖含量测定.食品研究与开发,201738(21):146-149.
[16]Zhang TX,Wang B,Zhang X,et al.Study on the degradation of sugars in Lentinus edodes fermentation broth by Bacillus subtilis.Food Ind2017,38(1):184-187(in Chinese).张腾霄,王斌,张希,等.枯草芽孢杆菌对香菇发酵液糖类成分降解规律研究.食品工业,201738(1):184-187.
[17]Zhang JG,Chen XM,He XS.Kinetic model for exopolysaccharides(EPS)of Ganoderma lucidum in batch fermentation.Chin J Biotech,2007,23(6):1065-1070(in Chinese).张建国,陈晓明,贺新生.灵芝胞外多糖分批发酵动力学模型.生物工程学报,2007,23(6):1065-1070.
[18]Han LR,Cheng D,Wang LR,et al.Structure and immunomodulatory activity of extracellular polysaccharide from Grifola frondosa.Chin JBiotech,2016,32(5):648-656(in Chinese).韩丽荣,程代,王莉蕊,等.灰树花胞外多糖的结构及免疫调节活性.生物工程学报,2016,32(5):648-656.
[19]Sun XQ,Zhou DQ,Wang SS,et al.Study on enhancement of antioxidant activity of tofu whey by fermentation with three species of Lactobacillus.Food Res Dev,2017,38(18):1-6(in Chinese).孙晓琦,周德庆,王珊珊,等.3种乳酸菌发酵提高黄浆水抗氧化能力的研究.食品研究与开发,2017,38(18):1-6.
[20]Saiga A,Tanabe S,Nishimura T.Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment.J Agric Food Chem,2003,51(12):3661-3667.
[21]Bai SW,Tang C,Tian J,et al.Extraction and antioxidant activity of total flavonoids from Sea buckthorn pomace Food Sci,2015,36(10):59-64(in Chinese).白生文,汤超,田京,等.沙棘果渣总黄酮提取工艺及抗氧化活性分析.食品科学,2015,36(10):59-64.
[22]Li Y,Yang S,Wan DG.Advances and existed problems in studies on liquid fermentation of medicinal fungi.Chin Tradit Herb Drugs,2012,43(10):2066-2070(in Chinese).李羿,杨胜,万德光.药用真菌液体发酵研究进展及存在问题探讨.中草药,2012,43(10):2066-2070.
[23]Zhuang Y,Pan Y,Xie XM,et al.The origin,development and its advantage and potential of“the bi-directional solid fermentation”for medicinal fungi.Edible Fungi China,2007,26(2):3-6(in Chinese).庄毅,潘扬,谢小梅,等.药用真菌“双向发酵”的起源、发展及其优势与潜力.中国食用菌,2007,26(2):3-6.
[24]Zhou YF,Cai ZX,Qin EH,et al.Effects of selenium on metabolism and total antioxidation of Cordyceps militari hypha.China Brew,2009,28(4):48-51(in Chinese).周毅峰,蔡忠雄,秦恩华,等.硒对蛹虫草菌丝代谢和总抗氧化能力的影响.中国酿造,2009,28(4):48-51.
[25]Zheng X,Tao CY,Xu JL,et al.Effect of three kinds of auxin on the liquid fermentation of Cordyceps militaris.Edible Fungi China,2017,36(6):68-71(in Chinese).郑鑫,陶春雨,徐嘉潞,等.3种生长素对蛹虫草液体发酵的影响.中国食用菌,2017,36(6):68-71.
[2]Chen AH,Fan W,Zhang LR,et al.Optimization of extraction of Cordyceps militaris polysaccharides and the antioxidant activity.J Anhui Agric Sci,2018,46(19):175-178(in Chinese).陈安徽,范旺,张良睿,等.蛹虫草多糖的提取及抗氧化活性研究.安徽农业科学,2018,46(19):175-178.
[3]Liu HJ,Jiang N,Li JJ,et al.Study on extraction and purification of polysaccharides from Cordyceps militaris Acta Agric Jiangxi,2007,19(12):80-82(in Chinese).刘红锦,蒋宁,李建军,等.蛹虫草多糖提取及纯化工艺研究.江西农业学报,2007,19(12):80-82.
[4]Wu GH,Wang M.Separation,purification and the immunoregulation effect of polysaccharides in cultured Cordyceps militaris.Chin J Nat Med,2007,5(1):73-76(in Chinese).吴光昊,王旻.蛹虫草多糖的分离及免疫活性的研究.中国天然药物,2007,5(1):73-76.
[5]Zhang S,Zhang YJ,Shrestha B,et al.Ophiocordyceps sinensis and Cordyceps militaris:research advances,issues and perspectives.Mycosystema,2013,32(4):577-597(in Chinese).张姝,张永杰,Shrestha B,等.冬虫夏草菌和蛹虫草菌的研究现状、问题及展望.菌物学报,2013,32(4):577-597.
[6]He ZY,Wu TX.Effects of Rhizoma gastrodiae on the submerged fermentation of Polyporus frondosus.Sci Technol Food Ind,2011,32(1):184-186,189(in Chinese).贺宗毅,吴天祥.天麻对灰树花液体深层发酵的影响.食品工业科技,2011,32(1):184-186,189.
[7]Xu DD.Study on bidirectional fermentation of Morchella esculenta and protocorms of Dendrobium candidum[D].Dalian:Dalian Polytechnic University2015(in Chinese).徐丹丹.羊肚菌与铁皮石斛原球茎的双向发酵[D].大连:大连工业大学,2015.
[8]Li YL,Su YY,Miao M,et al.Studies on fermentation conditions for exopolysaccharides production by Cordyceps militaris.J Taishan Med College,2007,28(12):942-944(in Chinese).李艳玲,苏延友,苗苗,等.泰山蛹虫草菌产胞外多糖发酵工艺的研究.泰山医学院学报,2007,28(12):942-944.
[9]Ye WJ.Studies on optimization of fermentation conditions,structure and antioxidant activity of Cordyceps militaris polysaccharides[D].Wuhan:Huazhong Agricultural University,2014(in Chinese).叶文姣.蛹虫草多糖的发酵条件优化、结构及抗氧化活性研究[D].武汉:华中农业大学,2014.
[10]Wang YM,Zhu WX,An LG,et al.Optimized cultivation conditions and production efficiency of mycelium and lixiviated cordycepic polysaccharide of Cordyceps militaris.J Unive Jinan:Sci&Tech2010,24(2):148-151(in Chinese).王永敏,祝文兴,安利国,等.优化条件对蛹虫草菌丝体与胞外多糖得率研究.济南大学学报:自然科学版,2010,24(2):148-151.
[11]Yang Z.Antitumor research and clinical applications of bitter leaf.J Nanjing Univ Tradit Chin Med,2013,29(4):397-400(in Chinese).杨早.南非叶化学成分及药理作用研究进展.南京中医药大学学报,2013,29(4):397-400.
[12]Huang W,Li XQ,Liu J,et al.Effects of different hormone combinations on callus inducing of shoot explant from bitter leaf(Vernonia amygdallina Del).J Trop Biol,2017,8(4):419-423(in Chinese).黄伟,黎秀琼,刘娇,等.不同激素配比对南非叶茎愈伤组织诱导的影响.热带生物学报,20178(4):419-423.
[13]Audu SA,Taiwo AE,Ojuolape AR.A study review of documented phytochemistry of Vernonia amygdalina(family asteraceae)as the basis for pharmacologic activity of plant extract.J Natl Sci Res,2012,2(7):1-8.
[14]Chen CF,Xiang XY,Gu Q,et al.Submerged fermentation conditions for synthesis of extracellular polysaccharide by Inonotus obliquus.Chin Tradit Herbal Drugs,2007,38(3):358-361(in Chinese).陈才法,项小燕,顾琪,等.桦褐孔菌胞外多糖合成的深层发酵条件研究.中草药,2007,38(3):358-361.
[15]Meng FL,Song ZF,Liu XX,et al.Determination of cordycepin and total sugar in Cordyceps sinensis and Cordyceps militaris.Food Res Dev,2017,38(21):146-149(in Chinese).孟繁磊,宋志峰,刘笑笑,等.冬虫夏草与蛹虫草中虫草素和总糖含量测定.食品研究与开发,201738(21):146-149.
[16]Zhang TX,Wang B,Zhang X,et al.Study on the degradation of sugars in Lentinus edodes fermentation broth by Bacillus subtilis.Food Ind2017,38(1):184-187(in Chinese).张腾霄,王斌,张希,等.枯草芽孢杆菌对香菇发酵液糖类成分降解规律研究.食品工业,201738(1):184-187.
[17]Zhang JG,Chen XM,He XS.Kinetic model for exopolysaccharides(EPS)of Ganoderma lucidum in batch fermentation.Chin J Biotech,2007,23(6):1065-1070(in Chinese).张建国,陈晓明,贺新生.灵芝胞外多糖分批发酵动力学模型.生物工程学报,2007,23(6):1065-1070.
[18]Han LR,Cheng D,Wang LR,et al.Structure and immunomodulatory activity of extracellular polysaccharide from Grifola frondosa.Chin JBiotech,2016,32(5):648-656(in Chinese).韩丽荣,程代,王莉蕊,等.灰树花胞外多糖的结构及免疫调节活性.生物工程学报,2016,32(5):648-656.
[19]Sun XQ,Zhou DQ,Wang SS,et al.Study on enhancement of antioxidant activity of tofu whey by fermentation with three species of Lactobacillus.Food Res Dev,2017,38(18):1-6(in Chinese).孙晓琦,周德庆,王珊珊,等.3种乳酸菌发酵提高黄浆水抗氧化能力的研究.食品研究与开发,2017,38(18):1-6.
[20]Saiga A,Tanabe S,Nishimura T.Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment.J Agric Food Chem,2003,51(12):3661-3667.
[21]Bai SW,Tang C,Tian J,et al.Extraction and antioxidant activity of total flavonoids from Sea buckthorn pomace Food Sci,2015,36(10):59-64(in Chinese).白生文,汤超,田京,等.沙棘果渣总黄酮提取工艺及抗氧化活性分析.食品科学,2015,36(10):59-64.
[22]Li Y,Yang S,Wan DG.Advances and existed problems in studies on liquid fermentation of medicinal fungi.Chin Tradit Herb Drugs,2012,43(10):2066-2070(in Chinese).李羿,杨胜,万德光.药用真菌液体发酵研究进展及存在问题探讨.中草药,2012,43(10):2066-2070.
[23]Zhuang Y,Pan Y,Xie XM,et al.The origin,development and its advantage and potential of“the bi-directional solid fermentation”for medicinal fungi.Edible Fungi China,2007,26(2):3-6(in Chinese).庄毅,潘扬,谢小梅,等.药用真菌“双向发酵”的起源、发展及其优势与潜力.中国食用菌,2007,26(2):3-6.
[24]Zhou YF,Cai ZX,Qin EH,et al.Effects of selenium on metabolism and total antioxidation of Cordyceps militari hypha.China Brew,2009,28(4):48-51(in Chinese).周毅峰,蔡忠雄,秦恩华,等.硒对蛹虫草菌丝代谢和总抗氧化能力的影响.中国酿造,2009,28(4):48-51.
[25]Zheng X,Tao CY,Xu JL,et al.Effect of three kinds of auxin on the liquid fermentation of Cordyceps militaris.Edible Fungi China,2017,36(6):68-71(in Chinese).郑鑫,陶春雨,徐嘉潞,等.3种生长素对蛹虫草液体发酵的影响.中国食用菌,2017,36(6):68-71.