该文利用响应面法对冠突散囊菌(Eurotium cristatum)所产的胞外黑色素发酵条件进行优化并对黑色素稳定性进行研究。以不同培养温度、培养基pH、摇床转速为评价因素,产生的黑色素溶液吸光值为响应值,建立3因素3水平Box-Behnken响应面试验。结果表明,冠突散囊菌胞外黑色素最优发酵条件参数为转速184 r/min、培养温度 29 ℃、培养基pH 6.8,在该发酵条件下提取获得冠突散囊菌胞外黑色素4.27 g/L。所得黑色素在70 ℃以下耐热性强,Mg2+、Zn2+、Fe3+对其的稳定性有增强作用,该黑色素应避光保存,在使用过程中应避免与强氧化剂和还原剂接触。
In this study, response surface methodology was employed to optimize the fermentation conditions and the stability of extracellular melanin produced by Eurotium cristatum was also investigated. The three-factor three-level Box-Behnken response surface experiment was established via setting different culture temperatures, pH of the culture medium, and the speed of the shaker as evaluation factors and regarding the absorbance value of produced melanin solution as response value. The results showed that the optimal fermentation parameters of E. cristatum extracellular melanin were 184 r/min, 29 ℃ and pH 6.8. The yields of extracellular melanin reached 4.27 g/L under this condition. The obtained melanin exhibited strong heat resistance when the temperature was below 70 ℃. Metal ions such as Mg2+, Zn2+, Fe3+ enhanced the stability of extracellular melanin. The melanin should be stored away from light and avoid the contact with the strong oxidants and reducing agents during use.
[1] 吴凯为.冠突散囊菌发酵刺五加茶的制备及品质分析[D].锦州:渤海大学,2019.
[2] 邹敏敏,董其惠,黄彦,等.冠突散囊菌发酵黑毛茶提取液的研究[J].生物加工过程,2019,17(4):409-417.
[3] CHEN Kaijiang,ZHEN Zeng,HUANG Yahui,et al.Chemical compositions of Pu'er tea fermented by Eurotium cristatum and their lipid-lowering activity[J].Food Science and Technology,2018,98:204-211.
[4] YAO Yani,WU Mengyao, HUANG Yingjie,et al.Appropriately raising fermentation temperature beneficial to the increase of antioxidant activity and gallic acid content in Eurotium cristatum-fermented loose tea[J].LWT-Food Science and Technology,2017,82:248-254.
[5] 覃金球.冠突散囊菌发酵液对金黄色葡萄球菌及其生物被膜的作用[D].南宁:广西医科大学,2019.
[6] 袁超,袁丹丹,王淼,等.黑茶益生菌种“金花”菌基因组测序及分析[J].食品与药品,2016,18(4):259-262.
[7] 郑欣欣.茯砖茶中“金花”菌产孢机制及其功能性研究[D].西安:陕西科技大学,2015.
[8] 祝雷强,蒋立勤,金湖欣.天然黑色素的生理功效、提取及应用研究进展[J].中国食物与营养,2013,19(3):72-74.
[9] CHEN Jianyi,WANG Chenjiaozi, SHU Canwei,et al.Isolation and characterization of a melanin from Rhizoctonia solani, the causal agent of rice sheath blight[J].Journal of the Agricultural Chemical Society of Japan, 2015, 50 (11):2 951-2 954.
[10] ANDRZEJ S,POSTLETHWAITE A E.Skin under the sun:When melanin pigment meets vitamin D[J].Endocrinology,2015, 156(1):1-4.
[11] ŁUKASZ Ł.Antioxidant, antibacterial properties and the light barrier assessment of raw and purified melanins isolated from Citrullus lanatus (watermelon) seeds[J].Herba Polonica, 2018,64(2):25-36.
[12] MA Yinpeng,BAO Yihong,KONG Xianghui,et al.Optimization of melanin extraction from the wood ear medicinal mushroom, auricularia auricula-judae(Agaricomycetes), by response surface methodology and its antioxidant activities in vitro[J]. International journal of medicinal mushrooms,2018,20(11):1 087-1 095.
[13] BARCENA A,BRUNO M,GENNARO A,et al. Melanins from two selected isolates of Pseudocercospora griseola grown in-vitro: Chemical features and redox activity[J].Journal of Photochemistry and Photobiology B,2018,186(7):207-215.
[14] YE Ziyang,LU Ying,ZONG Shuai,et al.Structure, molecular modification and anti-tumor activity of melanin from Lachnum singerianum[J].Process Biochemistry,2019,76(9):203-212.
[15] SHI Fang,LI Jinglei,YE Ziyang,et al.Antitumor effects of melanin from Lachnum YM226 and its derivative in H22 tumor-bearing mice[J].Med Chem Comm,2018,9(6):1 059-1 068.
[16] LI Shenglan,YANG Liuqing, LI Jinglei,et al.Structure, molecular modification, and anti- radiation activity of melanin from Lachnum YM156 on ultraviolet B-induced injury in mice[J].Applied Biochemistry and Biotechnology,2019,188(2):555-567.
[17] OBERG F,HASEEB A,AHNFELT M,et al.Herbal melanin activates TLR4/NF-κB signaling pathway[J].Phyto-medicine,2009, 16(5):477-484.
[18] PUGH N D,BALACHANDRAN P,LATA H,et al.Melanin: Dietary mucosal immune modulator from Echinacea and other botanical supplements[J].International Immunopharmacology,2005,5(4):637-647.
[19] 雷敏.鱿鱼墨黑色素及黑色素铁生物活性的研究[D].青岛:中国海洋大学,2008.
[20] 张艳荣,王庆庆,刘通,等.木耳黑色素碱法提取工艺优化及表征[J].食品科学,2016,37(12):27-32.
[21] 吴世玉,孟祥敏,刘俊梅,等.林蛙卵黑色素的理化性质及其抗氧化活性研究[J].食品研究与开发,2018,39(14):25-29.
[22] CHENG Fuyuan, HSU Fengwen,CHANG Hishan,et al.Effect of different acids on the extraction of pepsin-solubilised collagen containing melanin from silky fowl feet[J].Food Chemistry,2009,113(2):563-567.
[23] SELVAKUMAR P,RAJASEKAR S,PERIASAMY K,et al.Isolation and characterization of melanin pigment from Pleurotus cystidiosus (telomorph of Antromycopsis macrocarpa)[J].World Journal of Microbiogy&Biotechnology,2008,24(10):2 125-2 131.
[24] 郭劲霞,邹自征,刘蓉,等.冠突散囊菌液体培养条件的正交优化[J].安徽农业科学,2015,43(27):246-247;250.
[25] 刘善鑫,刘素纯,李再贵.冠突散囊菌发酵燕麦对阿魏酸含量的影响[J].中国酿造,2018,37(6):165-169.
[26] 陈云兰.茯砖茶“金花菌”的分类鉴定及其对茯砖茶品质的影响[D].南京:南京农业大学,2004.
[27] 满丽莉,向殿军.响应面法优化枯草芽孢杆菌MX-6产纳豆激酶发酵条件[J].食品研究与开发,2019,40(21):214-219.
[28] 杨然,张笑雨,刘朋肖,等.耐热木聚糖酶菌株的筛选鉴定及发酵条件分析[J].食品科学,2019,40(18):101-107.
