灵芝液态发酵高产胞外多糖菌株筛选及多糖特性分析
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摘要
对10株灵芝菌株发酵菌丝体生物量、胞内多糖含量、胞外粗多糖得率及其多糖含量和单糖组成特征进行了系统分析。结果表明,随着培养时间的延长,10株菌株的生物量均有不同程度增加,但不同菌株胞内多糖含量和胞外粗多糖得率变化趋势有所不同。在培养至7d时,G0023和G0160菌株胞外粗多糖得率均较高,分别为3.02g/L和3.14g/L,其多糖含量分别达到了84.11%和91.03%,可作为发酵高产胞外多糖的优良菌株。分级醇沉胞外多糖分析结果表明,各菌株胞外液20%乙醇沉淀所得20E组分的得率和多糖含量均高于50%乙醇沉淀所得的50E组分,说明胞外液中主要以大分子量多糖为主。20E主要由葡萄糖组成,含有少量木糖和甘露糖;50E主要由葡萄糖和甘露糖组成,含有少量木糖和半乳糖。胞外液表观粘度随剪切速率变化曲线分析结果显示,各菌株胞外液均呈现剪切变稀非牛顿流体特性,其表观粘度大小与菌株对应20E组分的得率及多糖含量呈正相关。
Ten strains of Ganoderma lingzhi were cultured by liquid fermentation to compare the mycelial biomass, the yields and content of extracellular polysaccharides, and monosaccharide composition characteristics. Results indicated that the mycelial biomass increased with extending culture period, however,different strains showed a difference in the content of intracellular polysaccharide and the yields of extracellular polysaccharide. Extracellular crude polysaccharide yields of two strains, G0023 and G0160,reached 3.02 and 3.14 g/L in seven days, and polysaccharide content was 84.11% and 91.03%, respectively.G0023 and G0160 could be regarded as excellent strains for producing extracellular polysaccharides. Analysis of ethanol fractional precipitated extracellular polysaccharides showed that the yields and polysaccharide content of 20 E fractions(precipitated with 20% ethanol) were higher than those of 50 E fractions(precipitated with 50% ethanol), indicating that the fermentation broth mainly contained high molecular weight polysaccharide fractions. 20 E fractions were composed principally of glucose with small amount of xylose and mannose. 50 E fractions mainly contained glucose and mannose together with small amount of xylose and galactose. Dependence of the apparent viscosity on the shear rate analysis for culture broth of different strains showed that culture broth exhibited non-Newtonian fluid and shear-thinning behavior. The values of apparent viscosity were positively correlated to the yields and polysaccharide content of 20 E.
Ten strains of Ganoderma lingzhi were cultured by liquid fermentation to compare the mycelial biomass, the yields and content of extracellular polysaccharides, and monosaccharide composition characteristics. Results indicated that the mycelial biomass increased with extending culture period, however,different strains showed a difference in the content of intracellular polysaccharide and the yields of extracellular polysaccharide. Extracellular crude polysaccharide yields of two strains, G0023 and G0160,reached 3.02 and 3.14 g/L in seven days, and polysaccharide content was 84.11% and 91.03%, respectively.G0023 and G0160 could be regarded as excellent strains for producing extracellular polysaccharides. Analysis of ethanol fractional precipitated extracellular polysaccharides showed that the yields and polysaccharide content of 20 E fractions(precipitated with 20% ethanol) were higher than those of 50 E fractions(precipitated with 50% ethanol), indicating that the fermentation broth mainly contained high molecular weight polysaccharide fractions. 20 E fractions were composed principally of glucose with small amount of xylose and mannose. 50 E fractions mainly contained glucose and mannose together with small amount of xylose and galactose. Dependence of the apparent viscosity on the shear rate analysis for culture broth of different strains showed that culture broth exhibited non-Newtonian fluid and shear-thinning behavior. The values of apparent viscosity were positively correlated to the yields and polysaccharide content of 20 E.
引文
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Fang Q,Zhong JJ,2002.Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum.Process Biochemistry,37(7):769-774
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Li YQ,Fang L,Zhang KC,2007.Structure and bioactivities of a galactose rich extracellular polysaccharide from submergedly cultured Ganoderma lucidum.Carbohydrate Polymers,68(2):323-328
Liu YF,Zhang JS,Tang QJ,Yang Y,Xia YM,Zhou S,Wu D,Zhang Z,Dong LL,Cui SW,2016.Rheological properties ofβ-d-glucan from the fruiting bodies of Ganoderma lucidum.Food Hydrocolloids,58:120-125
Nie SP,Zhang H,Li WJ,Xie MY,2013.Current development of polysaccharides from Ganoderma:isolation,structure and bioactivities.Bioactive Carbohydrates and Dietary Fibre,1(1):10-20
Russell R,Paterson M,2006.Ganoderma-a therapeutic fungal biofactory.Phytochemistry,67:1985-2001
Su PF,Tang QJ,Xu JF,Chen S,Wang YL,Zhao LB,Liu YF,2018.Study on the physicochemical properties and rheological properties of Nostoc sphaeroides Kütz polysaccharide.Science and Technology of Food Industry,39(14):39-43(in Chinese)
Tang YJ,Zhong JJ,2002.Fed-batch fermentation of Ganoderma lucidum for hyperproduction of polysaccharide and ganoderic acid.Enzyme and Microbial Technology,31(1-2):20-28
Wang GR,Feng J,Feng N,Tang CH,Zhang JS,2016.Screening and kinetic study of high-yield intracellular polysaccharide strains of Ganoderma lucidum in submerged fermentation.Acta Agriculture Shanghai,32(6):10-17(in Chinese)
Wang L,Su HY,Wang ZL,Ma PC,2008.Mycelial growth rates,biomass yield and polysaccharide content of eight Ganoderma strains.Acta Edulis Fungi,15(4):57-60(in Chinese)
Xu JW,Zhao W,Zhong JJ,2010.Biotechnological production and application of ganoderic acids.Applied Microbiololgy and Biotechnology,87:457-466
Yu ZJ,Chen CH,Zhao JY,2012.Optimization of conditions for testing polysaccharide content of edible fungi by DNSmethod.Jiangsu Agricultural Sciences,40(1):259-260(in Chinese)
Zhang J,Shen J,Liu YJ,Xia YM,2015.Constituents and structures of sulfated extracellular polysaccharides from Ganoderma lucidum fermentation broth.Journal of Food Science and Biotechnology,34(5):547-553(in Chinese)
Zou YL,Gan CL,Li P,2018.Progress in modern pharmacology of Ganoderma lucidum polysaccharides.Strait Pharmaceutical Journal,30(8):28-30(in Chinese)
Zuo Q,Yang HF,Xing ZT,Han Y,Gu SH,2008.Determination of total sugar in fruit bodies of five edible mushroom species.Acta Edulis Fungi,15(4):57-61(in Chinese)
戴玉成,曹云,周丽伟,吴声华,2013.中国灵芝学名之管见.菌物学报,32(6):947-952
蒋俊,杨焱,乔彦茹,周帅,刘艳芳,张劲松,冯志勇,2014.猴头菌多糖脱色前后理化性质及生物活性的研究.菌物学报,33(1):78-86
苏攀峰,唐庆九,许剑锋,陈盛,王玉兰,赵立彬,刘艳芳,2018.葛仙米多糖理化性质和流变学特性的研究.食品工业科技,39(14):39-43
王国瑞,冯杰,冯娜,唐传红,张劲松,2016.灵芝液体深层发酵高产胞内多糖菌株的筛选及其动力学研究.上海农业学报,32(6):10-17
王磊,宿红艳,王仲礼,马鹏程,2008.八种灵芝菌株的比较及筛选.食用菌学报,15(4):57-60
余志坚,陈传红,赵晋宇,2012.DNS法检测食用菌多糖含量条件优化研究.江苏农业科学,40(1):259-260
张珏,沈洁,刘昱均,夏咏梅,2015.硫酸化发酵灵芝胞外多糖的组成与结构.食品与生物技术学报,34(5):547-553
邹玉莲,甘陈灵,李鹏,2018.灵芝多糖现代药理学研究进展.海峡药学,30(8):28-30
左琦,杨海锋,邢增涛,韩烨,顾赛红,2008.食用菌总糖含量测定方法的研究.食用菌学报,15(4):61-65
Dai YC,Cao Y,Zhou LW,Wu SH,2013.Notes on the nomenclature of the most widely cultivated Ganoderma species in China.Mycosystema,32(6):947-952(in Chinese)
Fang Q,Zhong JJ,2002.Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum.Process Biochemistry,37(7):769-774
Jiang J,Yang Y,Qiao YR,Zhou S,Liu YF,Zhang JS,Feng ZY,2014.Physicochemical properties and biological activities of decolored and not decolored polysaccharides from Hericium erinaceus.Mycosystema,33(1):78-86(in Chinese)
Li YQ,Fang L,Zhang KC,2007.Structure and bioactivities of a galactose rich extracellular polysaccharide from submergedly cultured Ganoderma lucidum.Carbohydrate Polymers,68(2):323-328
Liu YF,Zhang JS,Tang QJ,Yang Y,Xia YM,Zhou S,Wu D,Zhang Z,Dong LL,Cui SW,2016.Rheological properties ofβ-d-glucan from the fruiting bodies of Ganoderma lucidum.Food Hydrocolloids,58:120-125
Nie SP,Zhang H,Li WJ,Xie MY,2013.Current development of polysaccharides from Ganoderma:isolation,structure and bioactivities.Bioactive Carbohydrates and Dietary Fibre,1(1):10-20
Russell R,Paterson M,2006.Ganoderma-a therapeutic fungal biofactory.Phytochemistry,67:1985-2001
Su PF,Tang QJ,Xu JF,Chen S,Wang YL,Zhao LB,Liu YF,2018.Study on the physicochemical properties and rheological properties of Nostoc sphaeroides Kütz polysaccharide.Science and Technology of Food Industry,39(14):39-43(in Chinese)
Tang YJ,Zhong JJ,2002.Fed-batch fermentation of Ganoderma lucidum for hyperproduction of polysaccharide and ganoderic acid.Enzyme and Microbial Technology,31(1-2):20-28
Wang GR,Feng J,Feng N,Tang CH,Zhang JS,2016.Screening and kinetic study of high-yield intracellular polysaccharide strains of Ganoderma lucidum in submerged fermentation.Acta Agriculture Shanghai,32(6):10-17(in Chinese)
Wang L,Su HY,Wang ZL,Ma PC,2008.Mycelial growth rates,biomass yield and polysaccharide content of eight Ganoderma strains.Acta Edulis Fungi,15(4):57-60(in Chinese)
Xu JW,Zhao W,Zhong JJ,2010.Biotechnological production and application of ganoderic acids.Applied Microbiololgy and Biotechnology,87:457-466
Yu ZJ,Chen CH,Zhao JY,2012.Optimization of conditions for testing polysaccharide content of edible fungi by DNSmethod.Jiangsu Agricultural Sciences,40(1):259-260(in Chinese)
Zhang J,Shen J,Liu YJ,Xia YM,2015.Constituents and structures of sulfated extracellular polysaccharides from Ganoderma lucidum fermentation broth.Journal of Food Science and Biotechnology,34(5):547-553(in Chinese)
Zou YL,Gan CL,Li P,2018.Progress in modern pharmacology of Ganoderma lucidum polysaccharides.Strait Pharmaceutical Journal,30(8):28-30(in Chinese)
Zuo Q,Yang HF,Xing ZT,Han Y,Gu SH,2008.Determination of total sugar in fruit bodies of five edible mushroom species.Acta Edulis Fungi,15(4):57-61(in Chinese)
戴玉成,曹云,周丽伟,吴声华,2013.中国灵芝学名之管见.菌物学报,32(6):947-952
蒋俊,杨焱,乔彦茹,周帅,刘艳芳,张劲松,冯志勇,2014.猴头菌多糖脱色前后理化性质及生物活性的研究.菌物学报,33(1):78-86
苏攀峰,唐庆九,许剑锋,陈盛,王玉兰,赵立彬,刘艳芳,2018.葛仙米多糖理化性质和流变学特性的研究.食品工业科技,39(14):39-43
王国瑞,冯杰,冯娜,唐传红,张劲松,2016.灵芝液体深层发酵高产胞内多糖菌株的筛选及其动力学研究.上海农业学报,32(6):10-17
王磊,宿红艳,王仲礼,马鹏程,2008.八种灵芝菌株的比较及筛选.食用菌学报,15(4):57-60
余志坚,陈传红,赵晋宇,2012.DNS法检测食用菌多糖含量条件优化研究.江苏农业科学,40(1):259-260
张珏,沈洁,刘昱均,夏咏梅,2015.硫酸化发酵灵芝胞外多糖的组成与结构.食品与生物技术学报,34(5):547-553
邹玉莲,甘陈灵,李鹏,2018.灵芝多糖现代药理学研究进展.海峡药学,30(8):28-30
左琦,杨海锋,邢增涛,韩烨,顾赛红,2008.食用菌总糖含量测定方法的研究.食用菌学报,15(4):61-65