利用扁桃斑鸠菊叶发酵高产粗毛纤孔菌胞外多糖的条件优化及其抗氧化活性研究
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摘要
粗毛纤孔菌胞外多糖是粗毛纤孔菌液体发酵的重要活性代谢产物,但采用常规的发酵方法,粗毛纤孔菌胞外多糖的产量较低。为更好地获取粗毛纤孔菌胞外多糖,本文采用双向液体发酵的方法,通过向发酵培养基中添加适量的扁桃斑鸠菊叶粉末,来提高粗毛纤孔菌胞外多糖的产量,并对优化得到的胞外多糖抗氧化活性进行了研究。以发酵液中胞外多糖含量为指标,采用单因素实验和正交实验优化发酵条件;采用红外光谱对胞外多糖的结构特征进行分析;通过测定胞外多糖对ABTS、DPPH和羟基自由基的清除率来了解其抗氧化活性。结果表明,最优发酵条件为:扁桃斑鸠菊叶粉末添加量0.5g/L、发酵时间10d、pH 6.5、接种量5.0mL,在此条件下,粗毛纤孔菌胞外多糖的产量达到(2.34±0.25)mg/mL,与未添加扁桃斑鸠菊叶的空白组相比,其胞外多糖产量提高了约216.22%;红外分析与抗氧化活性实验结果表明,添加扁桃斑鸠菊叶后的胞外多糖与未添加扁桃斑鸠菊叶的胞外多糖红外主要吸收峰一致,并且对ABTS、DPPH以及羟基自由基清除能力相近。本研究结果表明扁桃斑鸠菊叶能够有效地提高粗毛纤孔菌胞外多糖的产量,为其他珍稀食药用菌胞外多糖的高效生产提供了新思路。
Exopolysaccharide(EPS) is an important kind of active metabolite of Inonotus hispidus in liquid fermentation. Usually, the yield of Inonotus hispidus EPS is low obtained from conventional fermentation. In this study, bi-directional liquid fermentation was adopted to promote the Inonotus hispidus EPS production by adding Vernonia amygdalina leaf powder to the fermentation medium. Using the content of EPS in fermentation broth as main index, the fermentation conditions were optimized through single factor and orthogonal test, and infrared spectroscopy was used to analyze the structural features of EPS. ABTS, DPPH,hydroxyl radical scavenging rates were measured to detect the antioxidant activity of EPS. The optimum fermentation conditions were as follows: addition of V. amygdalina dried leaf powder of 0.5 g/L,fermentation duration of 10 d, initial pH of 6.5, inoculum size of 5.0 mL. Under such a condition, the yield of Inonotus hispidus EPS can reach(2.34±0.25)mg/mL, increased by 216.22% as compared with the experimental control without addition of the leaf powder. Infrared analysis and antioxidant activity tests indicated the principal absorption peaks of the EPS product accorded with those of experimental control,and the ability of the EPS product to scavenge ABTS, DPPH and hydroxyl radicals was similar to that of control. The result of this research suggests that V. amygdalina leaf can enhance the production of Inonotus hispidus EPS effectively.
Exopolysaccharide(EPS) is an important kind of active metabolite of Inonotus hispidus in liquid fermentation. Usually, the yield of Inonotus hispidus EPS is low obtained from conventional fermentation. In this study, bi-directional liquid fermentation was adopted to promote the Inonotus hispidus EPS production by adding Vernonia amygdalina leaf powder to the fermentation medium. Using the content of EPS in fermentation broth as main index, the fermentation conditions were optimized through single factor and orthogonal test, and infrared spectroscopy was used to analyze the structural features of EPS. ABTS, DPPH,hydroxyl radical scavenging rates were measured to detect the antioxidant activity of EPS. The optimum fermentation conditions were as follows: addition of V. amygdalina dried leaf powder of 0.5 g/L,fermentation duration of 10 d, initial pH of 6.5, inoculum size of 5.0 mL. Under such a condition, the yield of Inonotus hispidus EPS can reach(2.34±0.25)mg/mL, increased by 216.22% as compared with the experimental control without addition of the leaf powder. Infrared analysis and antioxidant activity tests indicated the principal absorption peaks of the EPS product accorded with those of experimental control,and the ability of the EPS product to scavenge ABTS, DPPH and hydroxyl radicals was similar to that of control. The result of this research suggests that V. amygdalina leaf can enhance the production of Inonotus hispidus EPS effectively.
引文
Audu SA,Taiwo AE,Ojuolape AR,2012.A study review of documented phytochemistry of Vernonia amygdalina(family Asteraceae)as the basis for pharmacologic activity of plant extract.Journal of Natural Sciences Research,7(2):1-8
Bai SW,Tang C,Tian J,Yan HF,Xu XS,Fan HL,2015.Extraction and antioxidant activity of total flavonoids from sea buckthorn pomace.Food Science,36(10):59-64(in Chinese)
Cao CL,Han ML,Cui BK,Yu HY,2013.Antioxidant activities of different fractional extracts from fruiting bodies of three Phellinus species.Mycosystema,32:883-890(in Chinese)
Cui PW,Wu HJ,Tan ZY,Cui Y,Liu XZ,2013.Primary exploration of metabolic characteristics in Poria cocos fermentation system with adding botanical medicine factors.Journal of Traditional Chinese Medicine University of Hunan,33(7):37-40(in Chinese)
Cui BK,Dai YC,Yang H,2009.Notes on the medicinal fungus of Inonotus hispidus.Edible Fungi of China,28(4):6-7(in Chinese)
Cao CL,Peng F,Cui BK,2013.Chemical characterization and structure of exopolysaccharides from submerged culture of new medicinal mushroom from China,Phellinus mori(Higher Basidiomycetes).International Journal of Medicinal Mushrooms,15:57-69
Chen CF,Xiang XY,Gu Q,Zheng WF,Wei JC,2007.Submerged fermentation condition for synthesis of extracellular polysaccharide by Inonotus obliquus.Chinese Traditional and Herbal Drugs,38(3):358-361(in Chinese)
Dai YC,2012.Pathogenic wood-decaying fungi on woody plants in China.Mycosystema,31(4):493-509(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Huang W,Li XQ,Liu J,Wei Q,Wu JS,2017.Effects of different hormone combinations on callus inducing of shoot explant from bitter leaf(Vernonia amygdallina Del).Journal of Tropical Biology,8(4):419-423(in Chinese)
Jin SS,Liu X,Hou RL,Yu QM,Wang DT,Hu KH,Zheng MF,Fu JS,2018.Identification and biological activity of a medicinal Inonotus.Journal of Northwest A&F University(Natural Science Edition),46(12):1-9(in Chinese)
Liu X,Hou RL,Jin SS,Qi M,Hu KH,Zheng MF,Fu JS,2018.Molecular screening of medicinal fungus Inonotus hispidus and anti-breast cancer activities of its submerged fermentation broth.Mycosystema,37(2):215-225(in Chinese)
Li QJ,Bao HY,Bau T,Li Y,2018.Influence of Inonotus hispidus in hemorheology in rat model with blood stasis due to cold syndrome and analysis on spectral efficiency.Journal of Jilin University(Medicine Edition),2018(1):30-35(in Chinese)
Li Y,Yang S,Wan DG,2012.Advances and existed problems in studies on liquid fermentation of medicinal fungi.Chinese Traditional and Herbal Drugs,43(10):2066-2070(in Chinese)
Meng FL,Song ZF,Liu XX,Niu HH,Cai HM,Wei CY,2017.Determination of cordycepin and total sugar in Cordyceps sinensis and Cordyceps militaris.Food Research and Development,38(21):146-149(in Chinese)
Meng G,Zheng F,Liu HX,Si J,Cui BK,2016.Antioxidant activity of Inonotus weigelae during the liquid cultivation.Biotechnology Bulletin,32(9):140-148(in Chinese)
Saiga A,Tanabe S,Nishimura T,2003.Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment.Journal of Agricultural and Food Chemistry,51(12):3661-3667
Sun XQ,Zhou DQ,Wang SS,Ma YJ,Zhu LL,Zhao F,2017.Study on enhancement of antioxidant activity of tofu whey by fermentation with three species of Lactobacillus.Food Research and Development,38(18):1-6(in Chinese)
Xin YH,Liang B,Wang YX,Liu Y,Bai XJ,2017.Optimization of the Ganoderma Iucidum-Ginkgo biloba bi-directional liquid fermentation condition and antioxidation properties of its products.Mycosystema,36(10):1427-1435(in Chinese)
Yang Z,2013.Antitumor research and clinical applications of bitter leaf.Journal of Nanjing University of Traditional Chinese Medicine,29(4):397-400(in Chinese)
Yu HY,Cao CL,Cui BK,2012.Polyphenol content and antioxidant activities of Phellinus Ionicericola in liquid culture.Mycosystema,31:933-939(in Chinese)
Zan LF,Bao HY,Bau,Li Y,Zhu YP,2013.Chemical composition in fruiting body of Inonotus hispidus(Hymenochaetales,Basidiomycota).Mycosystema,32(1):150-156(in Chinese)
Zhuang Y,Pan Y,Xie XM,Zhang LY,2007.The origin development and its advantage and potential of“the bi-directional solid fermentation”for medicinal fungi.Edible Fungi of China,26(2):3-6(in Chinese)
Zhang TX,Wang B,Zhang X,Ren QY,2017.Study on the degradation of sugars in Lentinus edodes fermentation broth by Bacillus subtilis.The Food Industry,2017(1):184-187(in Chinese)
白生文,汤超,田京,闫宏斐,许晓莎,范惠玲,2015.沙棘果渣总黄酮提取工艺及抗氧化活性分析.食品科学,36(10):59-64
曹春蕾,韩美玲,崔宝凯,余海尤,2013.三种木层孔菌子实体不同溶剂提取物抗氧化活性的研究.菌物学报,32:883-890
崔培梧,吴红娟,谭朝阳,崔妍,刘小忠,2013.植物药-茯苓菌双向发酵代谢特征初探.湖南中医药大学学报,33(7):37-40
崔宝凯,戴玉成,杨宏,2009.药用真菌粗毛纤孔菌概述.中国食用菌,28(4):6-7
陈才法,项小燕,顾琪,郑维发,魏江春,2007.桦褐孔菌胞外多糖合成的深层发酵条件研究.中草药,38(3):358-361
戴玉成,2012.中国木本植物病原木材腐朽菌研究.菌物学报,31(4):493-509
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
黄伟,黎秀琼,刘娇,魏青,吴金山,2017.不同激素配比对扁桃斑鸠菊茎愈伤组织诱导的影响.热带生物学报,8(4):419-423
金珊珊,刘鑫,侯若琳,俞全明,王丹亭,胡开辉,郑明锋,傅俊生,2018.1株药用纤孔菌的鉴定及其生物活性.西北农林科技大学学报(自然科学版),46(12):1-9
刘鑫,侯若琳,金珊珊,戚梦,胡开辉,郑明锋,傅俊生,2018.药用真菌粗毛纤孔菌的分子甄别及其发酵液抗乳腺癌活性研究.菌物学报,37(2):215-225
李庆杰,包海鹰,图力古尔,李玉,2018.粗毛纤孔菌对寒凝血瘀证大鼠血液流变学的影响及谱效关系分析.吉林大学学报(医学版),2018(1):30-35
李羿,杨胜,万德光,2012.药用真菌液体发酵研究进展及存在问题探讨.中草药,43(10):2066-2070
孟繁磊,宋志峰,刘笑笑,牛红红,蔡红梅,魏春雁,2017.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,38(21):146-149
孟歌,郑飞,刘红霞,司静,崔宝凯,2016.锦带花纤孔菌液体培养过程中的抗氧化活性.生物技术通报,32(9):140-148
孙晓琦,周德庆,王珊珊,马玉洁,朱兰兰,赵峰,2017.3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,38(18):1-6
辛燕花,梁彬,王颖霞,刘阳,白晓静,2017.灵芝-银杏双向液体发酵条件优化及抗氧化的研究.菌物学报,36(10):1427-1435
杨早,2013.扁桃斑鸠菊化学成分及药理作用研究进展.南京中医药大学学报,29(4):397-400
余海尤,曹春蕾,崔宝凯,2012.忍冬木层孔菌液体培养过程中多酚含量及抗氧化活性研究.菌物学报,31:933-939
昝立峰,包海鹰,图力古尔,李玉,朱艳萍,2013.粗毛纤孔菌子实体化学成分.菌物学报,32(1):150-156
庄毅,潘扬,谢小梅,张李阳,2007.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,26(2):3-6
张腾霄,王斌,张希,任清云,2017.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,2017(1):184-187
Bai SW,Tang C,Tian J,Yan HF,Xu XS,Fan HL,2015.Extraction and antioxidant activity of total flavonoids from sea buckthorn pomace.Food Science,36(10):59-64(in Chinese)
Cao CL,Han ML,Cui BK,Yu HY,2013.Antioxidant activities of different fractional extracts from fruiting bodies of three Phellinus species.Mycosystema,32:883-890(in Chinese)
Cui PW,Wu HJ,Tan ZY,Cui Y,Liu XZ,2013.Primary exploration of metabolic characteristics in Poria cocos fermentation system with adding botanical medicine factors.Journal of Traditional Chinese Medicine University of Hunan,33(7):37-40(in Chinese)
Cui BK,Dai YC,Yang H,2009.Notes on the medicinal fungus of Inonotus hispidus.Edible Fungi of China,28(4):6-7(in Chinese)
Cao CL,Peng F,Cui BK,2013.Chemical characterization and structure of exopolysaccharides from submerged culture of new medicinal mushroom from China,Phellinus mori(Higher Basidiomycetes).International Journal of Medicinal Mushrooms,15:57-69
Chen CF,Xiang XY,Gu Q,Zheng WF,Wei JC,2007.Submerged fermentation condition for synthesis of extracellular polysaccharide by Inonotus obliquus.Chinese Traditional and Herbal Drugs,38(3):358-361(in Chinese)
Dai YC,2012.Pathogenic wood-decaying fungi on woody plants in China.Mycosystema,31(4):493-509(in Chinese)
Dai YC,Yang ZL,2008.A revised checklist of medicinal fungi in China.Mycosystema,27(6):801-824(in Chinese)
Huang W,Li XQ,Liu J,Wei Q,Wu JS,2017.Effects of different hormone combinations on callus inducing of shoot explant from bitter leaf(Vernonia amygdallina Del).Journal of Tropical Biology,8(4):419-423(in Chinese)
Jin SS,Liu X,Hou RL,Yu QM,Wang DT,Hu KH,Zheng MF,Fu JS,2018.Identification and biological activity of a medicinal Inonotus.Journal of Northwest A&F University(Natural Science Edition),46(12):1-9(in Chinese)
Liu X,Hou RL,Jin SS,Qi M,Hu KH,Zheng MF,Fu JS,2018.Molecular screening of medicinal fungus Inonotus hispidus and anti-breast cancer activities of its submerged fermentation broth.Mycosystema,37(2):215-225(in Chinese)
Li QJ,Bao HY,Bau T,Li Y,2018.Influence of Inonotus hispidus in hemorheology in rat model with blood stasis due to cold syndrome and analysis on spectral efficiency.Journal of Jilin University(Medicine Edition),2018(1):30-35(in Chinese)
Li Y,Yang S,Wan DG,2012.Advances and existed problems in studies on liquid fermentation of medicinal fungi.Chinese Traditional and Herbal Drugs,43(10):2066-2070(in Chinese)
Meng FL,Song ZF,Liu XX,Niu HH,Cai HM,Wei CY,2017.Determination of cordycepin and total sugar in Cordyceps sinensis and Cordyceps militaris.Food Research and Development,38(21):146-149(in Chinese)
Meng G,Zheng F,Liu HX,Si J,Cui BK,2016.Antioxidant activity of Inonotus weigelae during the liquid cultivation.Biotechnology Bulletin,32(9):140-148(in Chinese)
Saiga A,Tanabe S,Nishimura T,2003.Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment.Journal of Agricultural and Food Chemistry,51(12):3661-3667
Sun XQ,Zhou DQ,Wang SS,Ma YJ,Zhu LL,Zhao F,2017.Study on enhancement of antioxidant activity of tofu whey by fermentation with three species of Lactobacillus.Food Research and Development,38(18):1-6(in Chinese)
Xin YH,Liang B,Wang YX,Liu Y,Bai XJ,2017.Optimization of the Ganoderma Iucidum-Ginkgo biloba bi-directional liquid fermentation condition and antioxidation properties of its products.Mycosystema,36(10):1427-1435(in Chinese)
Yang Z,2013.Antitumor research and clinical applications of bitter leaf.Journal of Nanjing University of Traditional Chinese Medicine,29(4):397-400(in Chinese)
Yu HY,Cao CL,Cui BK,2012.Polyphenol content and antioxidant activities of Phellinus Ionicericola in liquid culture.Mycosystema,31:933-939(in Chinese)
Zan LF,Bao HY,Bau,Li Y,Zhu YP,2013.Chemical composition in fruiting body of Inonotus hispidus(Hymenochaetales,Basidiomycota).Mycosystema,32(1):150-156(in Chinese)
Zhuang Y,Pan Y,Xie XM,Zhang LY,2007.The origin development and its advantage and potential of“the bi-directional solid fermentation”for medicinal fungi.Edible Fungi of China,26(2):3-6(in Chinese)
Zhang TX,Wang B,Zhang X,Ren QY,2017.Study on the degradation of sugars in Lentinus edodes fermentation broth by Bacillus subtilis.The Food Industry,2017(1):184-187(in Chinese)
白生文,汤超,田京,闫宏斐,许晓莎,范惠玲,2015.沙棘果渣总黄酮提取工艺及抗氧化活性分析.食品科学,36(10):59-64
曹春蕾,韩美玲,崔宝凯,余海尤,2013.三种木层孔菌子实体不同溶剂提取物抗氧化活性的研究.菌物学报,32:883-890
崔培梧,吴红娟,谭朝阳,崔妍,刘小忠,2013.植物药-茯苓菌双向发酵代谢特征初探.湖南中医药大学学报,33(7):37-40
崔宝凯,戴玉成,杨宏,2009.药用真菌粗毛纤孔菌概述.中国食用菌,28(4):6-7
陈才法,项小燕,顾琪,郑维发,魏江春,2007.桦褐孔菌胞外多糖合成的深层发酵条件研究.中草药,38(3):358-361
戴玉成,2012.中国木本植物病原木材腐朽菌研究.菌物学报,31(4):493-509
戴玉成,杨祝良,2008.中国药用真菌名录及部分名称的修订.菌物学报,27(6):801-824
黄伟,黎秀琼,刘娇,魏青,吴金山,2017.不同激素配比对扁桃斑鸠菊茎愈伤组织诱导的影响.热带生物学报,8(4):419-423
金珊珊,刘鑫,侯若琳,俞全明,王丹亭,胡开辉,郑明锋,傅俊生,2018.1株药用纤孔菌的鉴定及其生物活性.西北农林科技大学学报(自然科学版),46(12):1-9
刘鑫,侯若琳,金珊珊,戚梦,胡开辉,郑明锋,傅俊生,2018.药用真菌粗毛纤孔菌的分子甄别及其发酵液抗乳腺癌活性研究.菌物学报,37(2):215-225
李庆杰,包海鹰,图力古尔,李玉,2018.粗毛纤孔菌对寒凝血瘀证大鼠血液流变学的影响及谱效关系分析.吉林大学学报(医学版),2018(1):30-35
李羿,杨胜,万德光,2012.药用真菌液体发酵研究进展及存在问题探讨.中草药,43(10):2066-2070
孟繁磊,宋志峰,刘笑笑,牛红红,蔡红梅,魏春雁,2017.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,38(21):146-149
孟歌,郑飞,刘红霞,司静,崔宝凯,2016.锦带花纤孔菌液体培养过程中的抗氧化活性.生物技术通报,32(9):140-148
孙晓琦,周德庆,王珊珊,马玉洁,朱兰兰,赵峰,2017.3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,38(18):1-6
辛燕花,梁彬,王颖霞,刘阳,白晓静,2017.灵芝-银杏双向液体发酵条件优化及抗氧化的研究.菌物学报,36(10):1427-1435
杨早,2013.扁桃斑鸠菊化学成分及药理作用研究进展.南京中医药大学学报,29(4):397-400
余海尤,曹春蕾,崔宝凯,2012.忍冬木层孔菌液体培养过程中多酚含量及抗氧化活性研究.菌物学报,31:933-939
昝立峰,包海鹰,图力古尔,李玉,朱艳萍,2013.粗毛纤孔菌子实体化学成分.菌物学报,32(1):150-156
庄毅,潘扬,谢小梅,张李阳,2007.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,26(2):3-6
张腾霄,王斌,张希,任清云,2017.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,2017(1):184-187