纤维素酶-超声波协同提取黑木耳黑色素工艺及其抗氧化活性分析
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摘要
黑色素是黑木耳的主要活性成分之一,在黑木耳的药理活性上发挥着重要作用。为了提高黑木耳黑色素的提取得率,实验采用正交法和响应面法对纤维素酶-超声波协同提取黑木耳黑色素的提取工艺进行了优化,并对最优条件下提取的黑木耳黑色素体外抗氧化活性进行了分析。实验结果表明,纤维素酶-超声波协同提取黑木耳黑色素的最优条件为:酶添加量12mg,酶解温度40℃,酶解pH 5.0,酶解时间120min,NaOH浓度1.27mol/L,料液比1:40,超声功率300W,超声时间52min,超声温度60℃。在最优条件下,黑木耳黑色素提取得率可达到10.48%,相比于实验设置的未添加纤维素酶的超声波组黑色素提取得率提高了12.93%。抗氧化结果表明,采用纤维素酶-超声波协同提取的黑色素相比于未加纤维素酶提取的黑色素清除ABTS、DPPH和羟基自由基的能力更强。研究结果为黑木耳黑色素的高效提取及其产品的应用开发奠定了基础。
Melanin is one of the main active components of Auricularia heimuer and plays a significant role in pharmacological activities of Auricularia heimuer. Orthogonal and response surface methods were used to optimize the Auricularia heimuer melanin extraction process by cellulase and ultrasonic wave synergistic technology in order to improve the extraction yield of melanin. The antioxidant activity of the melanin product was analysed. Experimental results show that the optimum conditions for the synergistic extraction of melanin include enzyme additive quantity of 12 mg, enzymolysis temperature of 40°C, enzymolysis pH of5.0, enzymolysis duration of 120 min, NaOH concentration of 1.27 mol/L, solid-liquid ratio of 1:40, ultrasonic power of 300 W, ultrasonic duration of 52 min, and ultrasonic temperature of 60°C. Under such conditions,the extraction yielding rate of Auricularia heimuer melanin could reach 10.48%, 12.93% higher as compared with experimental group treated with ultrasound without addition of cellulase. The melanin product obtained from the cellulose-ultrasound synergistic process had more scavenging effects on ABTS, DPPH and hydroxyl radical as compared with that obtained from the extraction using only cellulose without ultrasonic treatment.
Melanin is one of the main active components of Auricularia heimuer and plays a significant role in pharmacological activities of Auricularia heimuer. Orthogonal and response surface methods were used to optimize the Auricularia heimuer melanin extraction process by cellulase and ultrasonic wave synergistic technology in order to improve the extraction yield of melanin. The antioxidant activity of the melanin product was analysed. Experimental results show that the optimum conditions for the synergistic extraction of melanin include enzyme additive quantity of 12 mg, enzymolysis temperature of 40°C, enzymolysis pH of5.0, enzymolysis duration of 120 min, NaOH concentration of 1.27 mol/L, solid-liquid ratio of 1:40, ultrasonic power of 300 W, ultrasonic duration of 52 min, and ultrasonic temperature of 60°C. Under such conditions,the extraction yielding rate of Auricularia heimuer melanin could reach 10.48%, 12.93% higher as compared with experimental group treated with ultrasound without addition of cellulase. The melanin product obtained from the cellulose-ultrasound synergistic process had more scavenging effects on ABTS, DPPH and hydroxyl radical as compared with that obtained from the extraction using only cellulose without ultrasonic treatment.
引文
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)
Cheng W,Qin JZ,Du JG,Zhang CH,2012.Optimization of ultrasonic-assisted enzymatic extraction of polysaccharide from Phellinus igniarius.Modern Food Science and Technology,28(6):662-666(in Chinese)
Fan JM,Xie CQ,Jia J,Xu Y,Zhang CX,2013.Opitimization of extraction processing on polysaccharides from Pleurotus eryngii by cellulase enzymolysis.Food Science and Technology,(3):192-196(in Chinese)
He CC,2011.Study on extraction,purification and properties of melanin from Auricularia auricular.Master Thesis,Qingdao Agricultural University,Qingdao.1-51(in Chinese)
Langfelder K,Streibel M,Jahn B,Haase G,Brakhage AA,2003.Biosynthesis of fungal melanins and their importance for human pathogenic fungi.Fungal Genetics and Biology,38(2):143-158
Li B,2011.Study on bacterial quorum sensing inhibition and antioxidant activity of melanin from Auricularia auricula.PhD Dissertation,Nanjing Agricultural University,Nanjing.1-113(in Chinese)
Li Q,Hou LH,Liu X,Li WL,Ma AM,2010.Identification and extraction technology of melanin from Auricularia auricula.Food Science,31(16):87-92(in Chinese)
Luan SY,2016.Study on Auricularia auricula polysaccharides lowering blood fat function and Auricularia auricula fat-lowering biscuit.Master Thesis,Northeast Forestry University,Harbin.1-46(in Chinese)
Manoharan S,Guillemin GJ,Abiramasundari RS,Essa MM,Akbar M,Akbar MD,2016.The role of reactive oxygen species in the pathogenesis of alzheimer's disease,parkinson's disease,and huntington's disease:a mini review.Oxidative Medicine and Cellular Longevity,2016(4):8590578
Miller AA,Silva TMD,Drummond GR,Sobey CG,Chrissobolis S,2014.Reactive oxygen species and cerebrovascular diseases.Berlin:Springer Berlin Heidelberg,1895-1924
Riley PA,1997.Melanin.International Journal of Biochemistry and Cell Biology,29(11):1235-1239
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 fermented with three species of Lactobacillus.Food Research and Development,38(18):1-6(in Chinese)
Tarangini K,Mishra S,2013.Production,characterization and analysis of melanin from isolated marine Pseudomonas sp.using vegetable waste.Research Journal of Engineering Sciences,2(5):40-46
Wu F,Dai YC,2015.Notes on the nomenclature of the Auricularia auricula-judae complex.Mycosystema,34(4):604-611(in Chinese)
Wu F,Yuan Y,He SH,Bandara AR,Hyde KD,Malysheva VF,Dai YC,2015.Global diversity and taxonomy of the Auricularia auricula-judae complex(Auriculariales,Basidiomycota).Mycological Progress,14:95
Wu GW,Li M,Liu Y,He XF,Bian YB,Huang W,2010.Studies on extraction of flavor nucletide from the stalk of Lentinus edodes with cellulose enzyme.China Condiment,35(12):41-43(in Chinese)
Wu ZC,2017.Preliminary study on the biosynthesis of main active ingredients of Auricularia auricula.Master Thesis,Wenzhou University,Wenzhou.1-61(in Chinese)
Zhang YR,Wang QQ,Liu T,Guo Z,2016.Alkaline extraction optimization and characterization of Auricularia auricula melanin.Food Science,37(12):27-32(in Chinese)
Zhou D,Shao L,Spitz DR,2014.Reactive oxygen species in normal and tumor stem cells.Advances in Cancer Research,122:1-67
Zou Y,2011.Fermentative production of melanin by Auricularia auricula and its functional properties.PhDDissertation,Nanjing Agricultural University,Nanjing.1-131(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
程伟,秦俊哲,杜军国,张春华,2012.桑黄多糖超声波协同纤维素酶法提取的工艺优化.现代食品科技,28(6):662-666
凡军民,谢春芹,贾君,徐银,张彩霞,2013.纤维素酶法提取杏鲍菇多糖工艺优化.食品科技,3:192-196
何长川,2011.黑木耳黑色素提取纯化及性质研究.青岛农业大学硕士论文,青岛.51
李斌,2011.黑木耳黑色素对细菌群体感应调控行为的抑制及其抗氧化活性的研究.南京农业大学博士论文,南京.113
李琦,侯丽华,刘鑫,李伟林,马爱民,2010.黑木耳黑色素鉴定及提取工艺优化.食品科学,31(16):87-92
栾淑莹,2016.黑木耳多糖降血脂功能及黑木耳降脂饼干的研究.东北林业大学硕士论文,哈尔滨.46
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
吴芳,戴玉成,2015.黑木耳复合群中种类学名说明.菌物学报,34(4):604-611
吴关威,李敏,刘吟,何小峰,边银丙,黄文,2010.纤维素酶法提取香菇柄中呈味核苷酸工艺研究.中国调味品,35(12):41-43
武志超,2017.木耳主要活性成分生物合成的初步研究.温州大学硕士论文,温州.61
张艳荣,王庆庆,刘通,郭中,2016.木耳黑色素碱法提取工艺优化及表征.食品科学,37(12):27-32
邹宇,2011.发酵法制备黑木耳色素及其功能特性研究.南京农业大学博士论文,南京.131
Cheng W,Qin JZ,Du JG,Zhang CH,2012.Optimization of ultrasonic-assisted enzymatic extraction of polysaccharide from Phellinus igniarius.Modern Food Science and Technology,28(6):662-666(in Chinese)
Fan JM,Xie CQ,Jia J,Xu Y,Zhang CX,2013.Opitimization of extraction processing on polysaccharides from Pleurotus eryngii by cellulase enzymolysis.Food Science and Technology,(3):192-196(in Chinese)
He CC,2011.Study on extraction,purification and properties of melanin from Auricularia auricular.Master Thesis,Qingdao Agricultural University,Qingdao.1-51(in Chinese)
Langfelder K,Streibel M,Jahn B,Haase G,Brakhage AA,2003.Biosynthesis of fungal melanins and their importance for human pathogenic fungi.Fungal Genetics and Biology,38(2):143-158
Li B,2011.Study on bacterial quorum sensing inhibition and antioxidant activity of melanin from Auricularia auricula.PhD Dissertation,Nanjing Agricultural University,Nanjing.1-113(in Chinese)
Li Q,Hou LH,Liu X,Li WL,Ma AM,2010.Identification and extraction technology of melanin from Auricularia auricula.Food Science,31(16):87-92(in Chinese)
Luan SY,2016.Study on Auricularia auricula polysaccharides lowering blood fat function and Auricularia auricula fat-lowering biscuit.Master Thesis,Northeast Forestry University,Harbin.1-46(in Chinese)
Manoharan S,Guillemin GJ,Abiramasundari RS,Essa MM,Akbar M,Akbar MD,2016.The role of reactive oxygen species in the pathogenesis of alzheimer's disease,parkinson's disease,and huntington's disease:a mini review.Oxidative Medicine and Cellular Longevity,2016(4):8590578
Miller AA,Silva TMD,Drummond GR,Sobey CG,Chrissobolis S,2014.Reactive oxygen species and cerebrovascular diseases.Berlin:Springer Berlin Heidelberg,1895-1924
Riley PA,1997.Melanin.International Journal of Biochemistry and Cell Biology,29(11):1235-1239
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 fermented with three species of Lactobacillus.Food Research and Development,38(18):1-6(in Chinese)
Tarangini K,Mishra S,2013.Production,characterization and analysis of melanin from isolated marine Pseudomonas sp.using vegetable waste.Research Journal of Engineering Sciences,2(5):40-46
Wu F,Dai YC,2015.Notes on the nomenclature of the Auricularia auricula-judae complex.Mycosystema,34(4):604-611(in Chinese)
Wu F,Yuan Y,He SH,Bandara AR,Hyde KD,Malysheva VF,Dai YC,2015.Global diversity and taxonomy of the Auricularia auricula-judae complex(Auriculariales,Basidiomycota).Mycological Progress,14:95
Wu GW,Li M,Liu Y,He XF,Bian YB,Huang W,2010.Studies on extraction of flavor nucletide from the stalk of Lentinus edodes with cellulose enzyme.China Condiment,35(12):41-43(in Chinese)
Wu ZC,2017.Preliminary study on the biosynthesis of main active ingredients of Auricularia auricula.Master Thesis,Wenzhou University,Wenzhou.1-61(in Chinese)
Zhang YR,Wang QQ,Liu T,Guo Z,2016.Alkaline extraction optimization and characterization of Auricularia auricula melanin.Food Science,37(12):27-32(in Chinese)
Zhou D,Shao L,Spitz DR,2014.Reactive oxygen species in normal and tumor stem cells.Advances in Cancer Research,122:1-67
Zou Y,2011.Fermentative production of melanin by Auricularia auricula and its functional properties.PhDDissertation,Nanjing Agricultural University,Nanjing.1-131(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
程伟,秦俊哲,杜军国,张春华,2012.桑黄多糖超声波协同纤维素酶法提取的工艺优化.现代食品科技,28(6):662-666
凡军民,谢春芹,贾君,徐银,张彩霞,2013.纤维素酶法提取杏鲍菇多糖工艺优化.食品科技,3:192-196
何长川,2011.黑木耳黑色素提取纯化及性质研究.青岛农业大学硕士论文,青岛.51
李斌,2011.黑木耳黑色素对细菌群体感应调控行为的抑制及其抗氧化活性的研究.南京农业大学博士论文,南京.113
李琦,侯丽华,刘鑫,李伟林,马爱民,2010.黑木耳黑色素鉴定及提取工艺优化.食品科学,31(16):87-92
栾淑莹,2016.黑木耳多糖降血脂功能及黑木耳降脂饼干的研究.东北林业大学硕士论文,哈尔滨.46
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
吴芳,戴玉成,2015.黑木耳复合群中种类学名说明.菌物学报,34(4):604-611
吴关威,李敏,刘吟,何小峰,边银丙,黄文,2010.纤维素酶法提取香菇柄中呈味核苷酸工艺研究.中国调味品,35(12):41-43
武志超,2017.木耳主要活性成分生物合成的初步研究.温州大学硕士论文,温州.61
张艳荣,王庆庆,刘通,郭中,2016.木耳黑色素碱法提取工艺优化及表征.食品科学,37(12):27-32
邹宇,2011.发酵法制备黑木耳色素及其功能特性研究.南京农业大学博士论文,南京.131