一株产黑色素黑蒜发酵菌的分离与鉴定
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摘要
黑色素是一类由植物或者微生物产生的色素,在食品、化妆品和医药领域具有广阔的应用前景。从黑蒜发酵96 h样品中筛选出1株产黑色素的细菌S8nyzx-1,对菌株进行形态学和分子生物学鉴定,采用紫外可见分光光谱和红外光谱扫描测定其所产黑色素特性,并进行短时耐高温能力测试。结果表明,在LB培养基上,菌落边缘较整齐,表面干燥,有褶皱,白色不透明,周围有少量棕色色素产生,16S r DNA测定核苷酸序列为1 419 bp,与枯草芽孢杆菌(Bacillus subtilis)同源性达99.8%。具有短时耐高温能力,可耐受93℃水浴10 min。由紫外分光光度法测得S8nyzx-1所产黑色素在370~390 nm左右有一个较强吸收峰;红外光谱分析黑色素具有吲哚结构的吸收峰,结合该菌可利用L-酪氨酸的特性,推测其为真黑色素。
Melanin is produced by plants or micro-organisms,and has broad application prospects in the food,cosmetics and medicine. Bacteria S8nyzx-1 that can produce melanin was isolated from black garlic of 96 h fermented. The strain was identified according to colony morphology and molecular biology. The structure characteristic of this melanin was identified using the techniques of UV and IR spectroscopy,and high temperature ability for a short time was determined. The results showed the colony was neat edges,white opaque,dry surface,wrinkled,and surrounded by a small amount of brown pigment on LB. 16 S r DNA fragment( 1 419 bp of size) was 99. 8% of similarity with Bacillus subtilis. The strain had a high ability of survive 10 min at 93 ℃ water bath. The melanin had strong absorption peak at 370- 390 nm through UV and with the indole structure through IR,which was considered for eumelanin according the characteristics of the bacteria can use L-tyrosine.
Melanin is produced by plants or micro-organisms,and has broad application prospects in the food,cosmetics and medicine. Bacteria S8nyzx-1 that can produce melanin was isolated from black garlic of 96 h fermented. The strain was identified according to colony morphology and molecular biology. The structure characteristic of this melanin was identified using the techniques of UV and IR spectroscopy,and high temperature ability for a short time was determined. The results showed the colony was neat edges,white opaque,dry surface,wrinkled,and surrounded by a small amount of brown pigment on LB. 16 S r DNA fragment( 1 419 bp of size) was 99. 8% of similarity with Bacillus subtilis. The strain had a high ability of survive 10 min at 93 ℃ water bath. The melanin had strong absorption peak at 370- 390 nm through UV and with the indole structure through IR,which was considered for eumelanin according the characteristics of the bacteria can use L-tyrosine.
引文
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[17]GONALVES R C,LISBOA H C,POMBEIRO-SPONCHIADO S R.Characterization of melanin pigment produced by Aspergillus nidulans[J].World Journal of Microbiology and Biotechnology,2012,28(4):1467-1474.
[18]YAO Zengyu,QI Jianhua,WANG Lihua.Isolation,fractionation and characterization of melanin-like pigments from chestnut(Castanea mollissima)shells[J].Journal of Food Science,2012,77(6):671-676.
[2]TARANGINI K,MISHRA S.Production of melanin by soil microbial isolate on fruit waste extract:two step optimization of key parameters[J].Biotechnology Reports,2014,4(12):139-146.
[3]GANESH K C,SAHU N,NARENDER R G,et al.Production of melanin pigment from Pseudomonas stutzeri isolated from red seaweed Hypnea musciformis[J].Letters in Applied Microbiology,2013,57(4):295-302.
[4]朱琳,商倩倩,游偲,等.一株产黑色素海洋细菌的筛选、鉴定及发酵条件优化[J].海洋环境科学,2011,30(5):686-688,698.ZHU L,SHANG Q Q,YOU C,et al.Isolation and culture condition optimization of a marine bacterium produced melanin[J].Marine Environmental Science,2011,30(5):686-688,698.
[5]张丽香,刘强,孙忠伟,等.一株产黑色素芽孢杆菌的分离与鉴定[J].华南农业大学学报,2010,31(2):63-67.
[6]卜利伟,邱瑞霞,黄雪松.黑蒜中呈色物质5-羟甲基糠醛的分离鉴定及其生成动力学[J].食品与发酵工业,2014,40(3):36-40.BU L W,QIU R X,HUANG X S.Isolation and identification of color material 5-hgdroxy methy furfural from black garlic and its kinetic formation[J].Food and Fermentation Industries,2014,40(3):36-40.
[7]邓振山,马娜娜,徐文梅,等.大蒜鳞茎中抗番茄灰霉病内生菌的筛选及其防治效果[J].西北农林科技大学学报(自然科学版),2012,40(5):50-56.
[8]崔北米,潘巧娜,张陪陪,等.大蒜内生细菌的分离及拮抗菌筛选与鉴定[J].西北植物学报,2008,28(11):2343-2348.
[9]钱存柔,黄仪秀.微生物学实验教程[M].北京:北京大学出版社,1999:53-57.
[10]惠明,马晓娜,贾洁,等.一株产聚谷氨酸芽孢杆菌的分离与鉴定[J].中国农业大学学报,2005,10(1):6-9,63.
[11]王斐,李俊霞,张丽香,等.产黑色素菌株发酵条件的研究[J].食品科技,2008,33(5):19-22.
[12]姬妍茹,石杰,刘宇峰,等.黑蒜生产过程中主要营养成分变化分析及工艺优化[J].食品工业科技,2015,36(5):360-364.JI Y R,SHI J,LIU Y F,et al.Changes analysis of main nutritional components in the production process of black garlic and process optimization[J].Science and Technology of Food Industry,2015,36(5):360-364.
[13]邓芹英,刘岚,邓慧敏.波谱分析教程[M].北京:科学出版社,2007:6-10.
[14]李建波,宋欣,曲音波.产胞外黑色素菌株的筛选[J].微生物学通报,2004,31(1):50-54.
[15]阎春兰,代金刚,王恒涛,等.土壤中一株产黑色素微生物的分离与鉴定[J].中南民族大学学报(自然科学版),2014,33(3):44-46.YAN C L,DAI J G,WANG H T,et al.Isolation and identification of one Melanin-Producing microorganism in Soil[J].Journal of South-Central University for Nationalities(Natural Science Edition),2014,33(3):44-46.
[16]姚增玉,赵忠,史清华,等.山杏种皮黑色素提取工艺研究[J].西北农林科技大学学报(自然科学版),2007,35(5):120-126.
[17]GONALVES R C,LISBOA H C,POMBEIRO-SPONCHIADO S R.Characterization of melanin pigment produced by Aspergillus nidulans[J].World Journal of Microbiology and Biotechnology,2012,28(4):1467-1474.
[18]YAO Zengyu,QI Jianhua,WANG Lihua.Isolation,fractionation and characterization of melanin-like pigments from chestnut(Castanea mollissima)shells[J].Journal of Food Science,2012,77(6):671-676.