真菌Simplicillium lanosoniveum色素的分离表征及培养基优化
|
摘要
对真菌Simplicillium lanosoniveum (DT06)胞外色素进行分离纯化、表征,特性研究以及培养基优化。通过酸化的乙酸乙酯萃取、蒸干、甲醇复溶、过滤得到高纯度真菌DT06胞外色素,并采用紫外-可见光谱和高效液相色谱分析对其表征,测定其抗氧化性和稳定性;然后基于Plackett-Burman试验设计法和响应面法对色素发酵培养基成分进行优化。实验结果表明:(1)DT06所产黄色色素是一种新色素,最大吸收峰为285 nm,为多羟基化合物;(2)DT06色素对DPPH自由基、ABTS自由基、超氧自由基及羟基自由基均有较强清除能力,在高温、强光及酸性条件下稳定,碱性条件下颜色变深、产生沉淀;(3)DT06色素合成的最佳培养基为:NaNO_3 0.68 g/L,K_2HPO_4 0.48 g/L,MgSO_4·7H_2O 0.037 g/L,CaCl_2·2H_2O 0.036 g/L,葡萄糖18 g/L,CS 14.5 mL/L,TM 1 mL/L,培养基优化后的色素产量是优化前的1.64倍。
Isolation and characterization of pigment from Simplicillium lanosoniveum(DT06)as well as its culture medium optimization were studied. The extracellular pigment of fungus DT06 was isolated by acidi?ed ethyl acetate,and re-dissolving with methanol. After puri?cation ?ltration,puri?ed products were characterized by ultravioletvisible spectroscopy and high performance liquid chromatography,and antioxidant activity and stability were tested.Furthermore,the pigment-production medium was optimized by Plackett-Burman design method and response surface methodology. The results showed that(1)The yellow pigment produced by fungus DT06 was a new pigment with a maximum absorption peak at 285 nm,probably a polyhydroxy compound;(2)DT06 pigment was able to scavenge DPPH,ABTS,superoxide and hydroxyl free radicals,and stable under high temperature,high irradiance and acid conditions;it became darken and precipitating under alkaline condition;(3)The optimized pigmentproduction medium is :NaNO3 0.68 g/L and K2 HPO4 0.48 g/L,MgSO4·7 H2 O 0.037 g/L,CaCl2·2 H2 O 0.036 g/L,glucose 18 g/L,CS 14.5 mL/L,TM 1 mL/L,and pigment yield in the optimized medium was 1.64 times before optimization.
Isolation and characterization of pigment from Simplicillium lanosoniveum(DT06)as well as its culture medium optimization were studied. The extracellular pigment of fungus DT06 was isolated by acidi?ed ethyl acetate,and re-dissolving with methanol. After puri?cation ?ltration,puri?ed products were characterized by ultravioletvisible spectroscopy and high performance liquid chromatography,and antioxidant activity and stability were tested.Furthermore,the pigment-production medium was optimized by Plackett-Burman design method and response surface methodology. The results showed that(1)The yellow pigment produced by fungus DT06 was a new pigment with a maximum absorption peak at 285 nm,probably a polyhydroxy compound;(2)DT06 pigment was able to scavenge DPPH,ABTS,superoxide and hydroxyl free radicals,and stable under high temperature,high irradiance and acid conditions;it became darken and precipitating under alkaline condition;(3)The optimized pigmentproduction medium is :NaNO3 0.68 g/L and K2 HPO4 0.48 g/L,MgSO4·7 H2 O 0.037 g/L,CaCl2·2 H2 O 0.036 g/L,glucose 18 g/L,CS 14.5 mL/L,TM 1 mL/L,and pigment yield in the optimized medium was 1.64 times before optimization.
引文
[1]Wang J.Research progress on natural pigments produced by microorganisms[J].Microbiology,2007.
[2]周波,吴吉林,朱明军,等.微生物黄色素的研究进展[J].微生物学通报,2010,37(9):1362-1368.
[3]Feng Y,Shao Y,Chen F.Monascus pigments.[J].Applied Microbiology&Biotechnology,2012,96(6):1421-1440.
[4]Caro Y,Anamale L,Fouillaud M,et al.Natural hydroxyanthraquinoid pigments as potent food grade colorants:an overview.[J].Natural Products&Bioprospecting,2012,2(5):174-193.
[5]Shier W T,Lao Y,Steele T W J,et al.Yellow pigments used in rapid identification of aflatoxin-producing Aspergillus strains are anthraquinones associated with the aflatoxin biosynthetic pathway[J].Bioorganic Chemistry,2005,33(6):426-438.
[6]周静,刘姝,胡晟源,等.一株产黄色素海洋真菌的分离、鉴定及黄色素特性的研究[J].食品工业科技,2017,38(10):217-220.
[7]Li D L,Li X M,Wang B G.Natural anthraquinone derivatives from a marine mangrove plant-derived endophytic fungus Eurotium rubrum:structural elucidation and DPPH radical scavenging activity.[J].J Microbiol Biotechnol,2009,19(7):675-680.
[8]Dong Q L,Lin T Y,Xing X Y,et al.Identification of a symbiotic fungus from blue-green alga and its extracellular polysaccharide[J].Letters in Applied Microbiology,2014,58(4):303-310.
[9]Dong Q,Dong R,Xing X,et al.A new antibiotic produced by the cyanobacterium-symbiotic fungus Simplicillium lanosoniveum[J].Natural Product Research,2017(4):1.
[10]董庆霖,李彤彤,邢向英,等.真菌Simplicillium lanosoniveum所产多糖分散性研究[J].食品科技,2018(6).
[11]苗佳旭.真菌DT06抗生素的分离纯化及理化性质初步研究[D].河北工业大学,2015.
[12]Sharmila G,Nidhi B,Muthukumaran C.Sequential statistical optimization of red pigment production by Monascus purpureus,(MTCC 369)using potato powder[J].Industrial Crops&Products,2013,44(2):158-164.
[13]张馨,曹红,李春,等.产紫青霉菌的发酵条件及色素稳定性、安全性的研究[J].中国食品添加剂,2016(2):68-74.
[14]Górna?P,?nēE,Siger A,et al.Sea buckthorn(Hippophae rhamnoides,L.)leaves as valuable source of lipophilic antioxidants:The effect of harvest time,sex,drying and extraction methods[J].Industrial Crops&Products,2014,60(1):1-7.
[15]Miller N J,Castelluccio C,Tijburg L,et al.The antioxidant properties of theaflavins and their gallate esters-radical scavengers or metal chelators[J].Febs Letters,1996,392(1):40.
[16]Li J,Min Z,Zheng T.The in vitro antioxidant activity of lotus germ oil from supercritical fluid carbon dioxide extraction[J].Food Chemistry,2009,115(3):939-944.
[17]Su D,Cheng Y,Liu M,et al.Comparision of piceid and resveratrol in antioxidation and antiproliferation activities in vitro[J].Plos One,2013,8(1):e54505.
[2]周波,吴吉林,朱明军,等.微生物黄色素的研究进展[J].微生物学通报,2010,37(9):1362-1368.
[3]Feng Y,Shao Y,Chen F.Monascus pigments.[J].Applied Microbiology&Biotechnology,2012,96(6):1421-1440.
[4]Caro Y,Anamale L,Fouillaud M,et al.Natural hydroxyanthraquinoid pigments as potent food grade colorants:an overview.[J].Natural Products&Bioprospecting,2012,2(5):174-193.
[5]Shier W T,Lao Y,Steele T W J,et al.Yellow pigments used in rapid identification of aflatoxin-producing Aspergillus strains are anthraquinones associated with the aflatoxin biosynthetic pathway[J].Bioorganic Chemistry,2005,33(6):426-438.
[6]周静,刘姝,胡晟源,等.一株产黄色素海洋真菌的分离、鉴定及黄色素特性的研究[J].食品工业科技,2017,38(10):217-220.
[7]Li D L,Li X M,Wang B G.Natural anthraquinone derivatives from a marine mangrove plant-derived endophytic fungus Eurotium rubrum:structural elucidation and DPPH radical scavenging activity.[J].J Microbiol Biotechnol,2009,19(7):675-680.
[8]Dong Q L,Lin T Y,Xing X Y,et al.Identification of a symbiotic fungus from blue-green alga and its extracellular polysaccharide[J].Letters in Applied Microbiology,2014,58(4):303-310.
[9]Dong Q,Dong R,Xing X,et al.A new antibiotic produced by the cyanobacterium-symbiotic fungus Simplicillium lanosoniveum[J].Natural Product Research,2017(4):1.
[10]董庆霖,李彤彤,邢向英,等.真菌Simplicillium lanosoniveum所产多糖分散性研究[J].食品科技,2018(6).
[11]苗佳旭.真菌DT06抗生素的分离纯化及理化性质初步研究[D].河北工业大学,2015.
[12]Sharmila G,Nidhi B,Muthukumaran C.Sequential statistical optimization of red pigment production by Monascus purpureus,(MTCC 369)using potato powder[J].Industrial Crops&Products,2013,44(2):158-164.
[13]张馨,曹红,李春,等.产紫青霉菌的发酵条件及色素稳定性、安全性的研究[J].中国食品添加剂,2016(2):68-74.
[14]Górna?P,?nēE,Siger A,et al.Sea buckthorn(Hippophae rhamnoides,L.)leaves as valuable source of lipophilic antioxidants:The effect of harvest time,sex,drying and extraction methods[J].Industrial Crops&Products,2014,60(1):1-7.
[15]Miller N J,Castelluccio C,Tijburg L,et al.The antioxidant properties of theaflavins and their gallate esters-radical scavengers or metal chelators[J].Febs Letters,1996,392(1):40.
[16]Li J,Min Z,Zheng T.The in vitro antioxidant activity of lotus germ oil from supercritical fluid carbon dioxide extraction[J].Food Chemistry,2009,115(3):939-944.
[17]Su D,Cheng Y,Liu M,et al.Comparision of piceid and resveratrol in antioxidation and antiproliferation activities in vitro[J].Plos One,2013,8(1):e54505.