一株生产有机硒菌的筛选鉴定及有机硒转化能力测定
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摘要
从烟草废弃物中分离出一株能大量生产有机硒的微生物菌株。采用细菌分离筛选方法选育出单菌种,用无机硒配制发酵液,进行有机硒转化效果试验。用光学显微镜观察菌种形态特征,根据《伯杰氏细菌鉴定手册》进行一系列生理化学反应观察其生理生化特征。应用基因测序技术,确定物种归属。结果表明,该菌为芽孢杆菌属枯草芽孢杆菌(Bacillus subtilis)种的新菌株。该菌可将700 mg/L亚硒酸钠全部转化为水溶性有机硒,用于水稻根外喷施,水稻对该发酵液发酵生产的有机硒利用率最高达64.12%,稻米中总硒含量最高达0.432 mg/kg,有机硒占总硒的比例最高达84.9%。
A microbial strain was separated from the tobacco waste,which could product mass organic selenium. By bacterial isolation and screening method, breeding a single species,the conversion effect of organic selenium was tested with the fermentation liquid of inorganic selenium. The morphological characteristics of bacteria were observed by optical microscopy. The physiological and biochemical characteristics were observed according to Bergey 's Manual of a series of physiological and chemical reaction. The species belonging was determined by application of gene sequencing technology. The results showed that the bacterium was a new subspecies of Bacillus subtilis. The bacteria could make 700 mg/L sodium selenite converted into water soluble organic selenium,used in rice root outside spraying. And the effects of different fermentation rice production had the highest utilization rate of organic selenium was 64.12%,and the total selenium content in rice was up to 0.432 mg/kg,and organic selenium percentage of total selenium was up to 84.9%.
A microbial strain was separated from the tobacco waste,which could product mass organic selenium. By bacterial isolation and screening method, breeding a single species,the conversion effect of organic selenium was tested with the fermentation liquid of inorganic selenium. The morphological characteristics of bacteria were observed by optical microscopy. The physiological and biochemical characteristics were observed according to Bergey 's Manual of a series of physiological and chemical reaction. The species belonging was determined by application of gene sequencing technology. The results showed that the bacterium was a new subspecies of Bacillus subtilis. The bacteria could make 700 mg/L sodium selenite converted into water soluble organic selenium,used in rice root outside spraying. And the effects of different fermentation rice production had the highest utilization rate of organic selenium was 64.12%,and the total selenium content in rice was up to 0.432 mg/kg,and organic selenium percentage of total selenium was up to 84.9%.
引文
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[2]CHANG W C,CHEN S H,WU L F,et al.Cytoprotective effect of reduced glutathione in arsenical-induced endothelial cell injury[J].Toxicology,1991,69(1):101-110.
[3]徐厚恩.卫生毒理学基础[M].北京:北京医科大学,中国协和医科大学联合出版社,1991.
[4]WHANGER P D.China,a country with both selenium deficiency and toxicity:Some thoughts and impressions[J].Nutr,1989,119:1236-1239.
[5]徐兵河,孙燕.微量元素硒的防癌抗癌作用[J].中国新药杂志,1998,7(3):176-179.
[6]HATFIELD D L,BERRY M J,GLADYSHEV V N.Selenium:Its Molecular Biology and Role in Human Health[M].2nd ed.New York:Springer Science+Business Media,LLC,2006.
[7]中国科学院地理研究所环境与地方病研究组.我国低硒带与克山病、大骨节病病因关系的研究[J].环境科学,1986,7(4):89-93.
[8]黄开勋,徐辉碧.硒的化学、生物化学及其在生命科学中的应用[M].第二版.武汉:华中科技大学出版社,2009.
[9]夏弈明.中国人体硒营养研究回顾[J].营养学报,2011,33(4):329-334.
[10]夏弈明,HILL K E,李平,等.中国成人硒需要量研究[J].营养学报,2011,33(2):109-113.
[11]葛可佑.中国营养科学全书[M].北京:人民卫生出版社,2004.
[12]李华为,赵素云.硒多糖的抗氧化活性及与过氧化氢酶协同作用的研究[J].分析化学,2010,9(38):1256-1260.
[13]刘琼,姜亮.硒蛋白的分子生物学及与疾病的关系[J].化学进展,2009,15(15):819-830.
[14]王新风,陶刚琴,陈媛媛.富硒酵母的培养和硒核酸提取的研究[J].中国食品学报,2006,6(6):52-57.
[15]中国富硒农业产业技术创新联盟.中国富硒农业发展蓝皮书[M].北京:中国农业科学技术出版社,2015.
[16]王世平,周玉岩,滕冰,等.酵母中有机硒转化的研究[J].东北农业大学学报,1997,28(4):378-383.
[17]KULP T R,PRATT L M.Speciation and weathering of selenium in Upper Cretaceous chalk and shale from South Dakota and Wyoming,USA[J].Geochemical Et Cosmo chemical Acta,2004,68(18):3687-3701.
[18]NOGUCHI H,UCHINO M,SHIVA O,et al.Bacillus Vietnamensis sp.nov.,a moderately halotolerant,aerobic,endospermforming bacterium isolated from Vietnamese fish sauce[J].Syst Evol Micr,2004,16(54):2117-2120.
[19]YOON J H,KIM I G,KANG K H,et al.Bacillus marisflavi sp.nov.and Bacillus aquimaris sp.nov.,isolated from sea water of a tidal flat of the Yellow Sea in Korea[J]Syst Evol Micr,2003,53:1297-1303.
[20]于超,郭海勇,魏嘉良,等.16S~23S r RNA基因序列在细菌鉴定中的应用[J].中国畜牧兽医,2012,39(2):57-60.