黄曲霉毒素B_1降解菌株的筛选及鉴定
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摘要
该研究以香豆素为唯一碳源,从豆类发酵食品、土壤、动物肠道及其内容物中筛选黄曲霉毒素B_1(AFB_1)降解菌株;然后通过复筛,从中筛选出AFB_1降解能力较好的菌株,并对其降解作用与吸附作用进行区分;最后,通过形态观察、生理生化试验及分子生物学技术对其进行鉴定。结果表明,共筛选得到9株AFB_1降解菌株,其中菌株YC2的AFB_1降解能力最强,AFB_1降解率达到90.7%,并确定菌株YC2通过降解作用去除AFB_1。最后,菌株YC2被鉴定为枯草芽孢杆菌(Bacillus subtilis)。
Using coumarin as sole carbon source, the aflatoxin B_1(AFB_1) degrading strains were screened from fermented soybean food, soil, animal intestine and its contents. Then, the strain with AFB_1 degrading ability was screened out by rescreening tests, and the degradation and adsorption were distinguished. Finally, the strain was identified by morphology observation, physiological and biochemical tests and molecular biological technique.The results showed that nine AFB_1 degrading strains were obtained. Among them, the AFB_1 degrading ability of the strain YC2 was the strongest, the degradation rate of AFB_1 reached 90.7%, and it was determined that strain YC2 could remove AFB_1 through degradation. Finally, the strain YC2 was identified as Bacillus subtilis.
Using coumarin as sole carbon source, the aflatoxin B_1(AFB_1) degrading strains were screened from fermented soybean food, soil, animal intestine and its contents. Then, the strain with AFB_1 degrading ability was screened out by rescreening tests, and the degradation and adsorption were distinguished. Finally, the strain was identified by morphology observation, physiological and biochemical tests and molecular biological technique.The results showed that nine AFB_1 degrading strains were obtained. Among them, the AFB_1 degrading ability of the strain YC2 was the strongest, the degradation rate of AFB_1 reached 90.7%, and it was determined that strain YC2 could remove AFB_1 through degradation. Finally, the strain YC2 was identified as Bacillus subtilis.
引文
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[2]计成,赵丽红.黄曲霉毒素生物降解的研究及前景展望[J].动物营养学报,2010(2):241-245.
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[9]关心,何剑斌,董双,等.黄曲霉毒素B1高效降解菌株的筛选鉴定及其降解[J].华中农业大学学报,2016,35(2):90-96.
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[11]李超波,李文明,杨文华,等.降解黄曲霉毒素微生物筛选中降解与吸附结合作用的区分[J].食品与发酵工业,2012,38(5):17-21.
[12]戴军,邵帅,王常高,等.产黄曲霉毒素B1降解酶菌株的筛选鉴定及发酵条件优化[J].工业微生物,2014,44(6):54-59.
[13]张志敏,王燕,毛勇,等.一株瘤胃黄曲霉毒素B1降解菌的分离鉴定及特性研究[J].动物医学进展,2014,35(8):49-53.
[14]计成.霉菌毒素与饲料食品安全[M].北京:化学工业出版社,2007:36-37.
[15]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:349-398.
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[19]BRADY D,STOLL A D,STARKE L,et al.Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae[J].Biotechnol Bioeng,1994,44(3):297-302.
[20]HASKARD C A,EL-NEZAMI H S,KANKAANP P E,et al.Surface binding of aflatoxin B1by lactic acid bacteria[J].Appl Environ Microbiol,2001,67(7):3086-3091.
[21]PELTONEN K,EL-NEZAMI H,HASKARD C,et al.Aflatoxin B1binding by dairy strains of lactic acid bacteria and bifidobacteria[J].JDairy Sci,2001,84(10):2152-2156.
[22]TURBIC A,AHOKAS J T,HASKARD C A,et al.Selective in vitro binding of dietary mutagens,individually or in combination,by lactic acid bacteria[J].Food Addit Contam,2002,19(2):144-152.
[2]计成,赵丽红.黄曲霉毒素生物降解的研究及前景展望[J].动物营养学报,2010(2):241-245.
[3]刘立芳.黄曲霉毒素的检测及其降解方法进展[J].中国酿造,2014,33(1):23-26.
[4]DAS C,MISHRA H N.In vitro degradation of aflatoxin B1in groundnut(Arachis hypogea)meal by horse radish peroxidase[J].LWT-Food Sci Technol,2000,33(4):308-312.
[5]王海鸥,陈守江,胡志超,等.花生黄曲霉毒素污染与控制[J].江苏农业科学,2015,43(1):270-273.
[6]PETCHKONGKAEW A,TAILLANDIER P,GASALUCK P,et al.Isolation of Bacillus spp.from Thai fermented soybean(Thua-nao):screening for aflatoxin B1and ochratoxin A detoxification[J].J Appl Microbiol,2008,104(5):1495-1502.
[7]TENIOLA O,ADDO P,BROST I,et al.Degradation of aflatoxin B1by cell-free extracts of Rhodococcus erythropolis and Mycobacterium fluoranthenivorans sp.nov.DSM44556T[J].Int J Food Microbiol,2005,105(2):111-117.
[8]张盼.生物法降解黄曲霉毒素B1的初步研究[D].北京:中国农业科学院,2015.
[9]关心,何剑斌,董双,等.黄曲霉毒素B1高效降解菌株的筛选鉴定及其降解[J].华中农业大学学报,2016,35(2):90-96.
[10]朱新贵,林捷.几种食品微生物降解黄曲霉毒素作用的研究[J].食品科学,2001,22(10):65-68.
[11]李超波,李文明,杨文华,等.降解黄曲霉毒素微生物筛选中降解与吸附结合作用的区分[J].食品与发酵工业,2012,38(5):17-21.
[12]戴军,邵帅,王常高,等.产黄曲霉毒素B1降解酶菌株的筛选鉴定及发酵条件优化[J].工业微生物,2014,44(6):54-59.
[13]张志敏,王燕,毛勇,等.一株瘤胃黄曲霉毒素B1降解菌的分离鉴定及特性研究[J].动物医学进展,2014,35(8):49-53.
[14]计成.霉菌毒素与饲料食品安全[M].北京:化学工业出版社,2007:36-37.
[15]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:349-398.
[16]布坎南R E,吉本斯N E.伯杰氏细菌鉴定手册(第八版)[M].北京:科学出版社,1984:729-830.
[17]LEE L S,DUNN J J,DELUCCA A J,et al.Role of lactone ring of aflatoxin B1in toxicity and mutagenicity[J].Experientia,1981,37(1):16-17.
[18]AZAB R M,TAWAKKOL W M,HAMAD A R M,et al.Detection and estimation of aflatoxin B1in feeds and its biodegradation by bacteria and fungi[J].Egypt J Nat Toxins,2005,2:39-56.
[19]BRADY D,STOLL A D,STARKE L,et al.Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae[J].Biotechnol Bioeng,1994,44(3):297-302.
[20]HASKARD C A,EL-NEZAMI H S,KANKAANP P E,et al.Surface binding of aflatoxin B1by lactic acid bacteria[J].Appl Environ Microbiol,2001,67(7):3086-3091.
[21]PELTONEN K,EL-NEZAMI H,HASKARD C,et al.Aflatoxin B1binding by dairy strains of lactic acid bacteria and bifidobacteria[J].JDairy Sci,2001,84(10):2152-2156.
[22]TURBIC A,AHOKAS J T,HASKARD C A,et al.Selective in vitro binding of dietary mutagens,individually or in combination,by lactic acid bacteria[J].Food Addit Contam,2002,19(2):144-152.