降解菌DDS-1产3-AC-DON氧化酶的酶学特性

doi:10.3864/j.issn.0578-1752.2013.11.007

中国农业科学 ›› 2013, Vol. 46 ›› Issue (11): 2240-2248.doi: 10.3864/j.issn.0578-1752.2013.11.007

降解菌DDS-1产3-AC-DON氧化酶的酶学特性

徐剑宏, 潘艳梅, 胡晓丹, 祭芳, 殷宪超, 史建荣

1.江苏省农业科学院食品质量安全与检测研究所/江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/农业部农产品质量安全控制技术与标准重点实验室/江苏省转基因安全评价公共服务中心，南京 210014
2.南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室，南京 210095

收稿日期:2012-12-04 出版日期:2013-06-01 发布日期:2013-04-01
通讯作者: 通信作者史建荣，Tel：025-84392001；E-mail：Jianrong63@126.com
作者简介:徐剑宏，Tel：025-84392001；E-mail：xujianhongnj@126.com；潘艳梅，E-mail：panyanmei-163-com@163.com。徐剑宏与潘艳梅为同等贡献作者
基金资助:
国家自然科学基金项目（30901006）、公益性行业（农业）科研专项（201303088）、江苏省农业科技创新基金（CX(11)2071）、江苏省科技项目（BE2010757）

Enzymatic Characteristics of 3-Acetyl Deoxynivalenol Oxidase by Devosia sp. DDS-1

XU Jian-Hong, PAN Yan-Mei, HU Xiao-Dan, JI Fang, YIN Xian-Chao, SHI Jian-Rong

1.Institute of Safety and Detection of Food Quality, Jiangsu Academy of Agricultural Sciences/Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base/Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality , Ministry of Agriculture/Jiangsu Center for GMO Evaluation and Detection, Nanjing 210014
2.Key Laboratory of Microbiological Engineering Agricultural Environment, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing 210095

Received:2012-12-04 Online:2013-06-01 Published:2013-04-01

摘要/Abstract

摘要： 【目的】研究脱氧雪腐镰刀菌烯醇（deoxynivalenol，DON）降解菌Devosia sp. DDS-1产生的3-AC-DON氧化酶的酶学特性，掌握DDS-1降解3-AC-DON到3-酮基-DON特性，为该酶的开发应用奠定理论基础。【方法】LG培养基培养DON毒素降解菌Devosia sp. DDS-1后，用PBS离心洗涤菌体后采用超声波破碎，用硫酸铵沉淀菌体破碎液获得粗酶液；然后在不同的酶反应体系中检测3-AC-DON氧化酶的酶活性。【结果】降解菌DDS-1产生3-AC-DON氧化酶的最适条件为：pH 7.0、25℃、培养36 h；该酶酶反应的最适温度为35℃，最适反应pH为7.0；该酶在20—40℃，pH 5.0—9.0的范围内能保持80%以上的酶活性；大多数金属离子在浓度为0.2—2 mmol•L-1对3-AC-DON氧化酶有促进作用；乙二胺四乙酸（EDTA）对该酶活性有抑制作用；酶的定域试验结果显示该酶为胞内酶；DDS-1产生的粗酶液能很好地降解小麦中的DON、3-AC-DON毒素。【结论】Devosia sp. DDS-1产生的3-AC-DON氧化酶的酶学特性研究表明，DDS-1产生的3-AC-DON氧化酶具有较好的温度和酸碱稳定性，该酶反应需要金属离子作为辅因子。

关键词: 3-AC-DON , 降解 , 徳沃斯氏菌 , 酶活性

Abstract: 【Objective】 The objective of this study is to investigate the enzymatic characteristics of 3-acetyl deoxynivalenol oxidase (3-AC-DON oxidase), understand the degradation characteristics of 3-acetyl deoxynivalenol by Devosia sp. DDS-1, and realize the application of the enzyme.【Method】After being cultured in LG medium , the strain DDS-1 was centrifuged and washed by PBS, and then broken by ultrasonic treatment. The crude enzyme of DDS-1 was obtained by ammonium sulfate precipitation. The enzymatic activity of 3-AC-DON oxidase was then detected in different enzyme reaction systems. 【Result】 The optimal 3-AC-DON oxidase production conditions for DDS-1 were 25℃, pH 7.0 and 36 hours in LG medium. The optimum reaction conditions for the 3-AC-DON oxidase in DDS-1 were 35℃, pH 7.0. The enzymatic activity could be maintained above 80% from 20 to 40℃ and in the range of pH 5.0 to 9.0. Most metal ions at a concentration from 0.2 to 2 mmol•L-1 could promote the enzymatic activity. Ethylenediamine tetraacetic acid (EDTA) inhibited the enzyme activity on the contrary. The crude enzyme produced by DDS-1 could degrade DON, 3-AC-DON in wheat.【Conclusion】The enzymatic characteristics of 3-AC-DON oxidase produced by Devosia sp. DDS-1 was studied in this paper. The results showed that Devosia sp. DDS-1 can produce 3-AC-DON oxidase with good temperature and pH stability, and the enzyme requires metal ions as cofactor.

Key words: 3-acetyl deoxynivalenol , degradation , Devosia sp. , enzymatic activity

徐剑宏, 潘艳梅, 胡晓丹, 祭芳, 殷宪超, 史建荣. 降解菌DDS-1产3-AC-DON氧化酶的酶学特性[J]. 中国农业科学, 2013, 46(11): 2240-2248.

XU Jian-Hong, PAN Yan-Mei, HU Xiao-Dan, JI Fang, YIN Xian-Chao, SHI Jian-Rong. Enzymatic Characteristics of 3-Acetyl Deoxynivalenol Oxidase by Devosia sp. DDS-1[J]. Scientia Agricultura Sinica, 2013, 46(11): 2240-2248.

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参考文献

[1]Jaksic S, Abramovic B, Jajic I, Balos M Z, Mihaljev Z, Despotovic V, Sojic D. Co-occurrence of fumonisins and deoxynivalenol in wheat and maize harvested in Serbia. Bulletin of Environmental Contamination and Toxicology, 2012, 89(3): 615-619.

[2]van der Fels-Klerx H J, de Rijk T C, Booij C J, Goedhart P W, Boers E A M, Zhao C, Waalwijk C, Mol H G J, van der Lee T A J. Occurrence of Fusarium head blight species and Fusarium mycotoxins in winter wheat in the Netherlands in 2009. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(11): 1716-1726.

[3]Lee T, Lee S H, Lee S H, Shin J Y, Yun J C, Lee Y W, Ryu J G. Occurrence of Fusarium mycotoxins in rice and its milling by-products in Korea. Journal of Food Protection, 2011, 74(7): 1169-1174.

[4]Mylona K, Sulyok M, Magan N. Relationship between environmental factors, dry matter loss and mycotoxin levels in stored wheat and maize infected with Fusarium species. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(7): 1118-1128.

[5]Edwards S G, Dickin E T, MacDonald S, Buttler D, Hazel C M, Patel S, Scudamore K A. Distribution of Fusarium mycotoxins in UK wheat mill fractions. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2011, 28(12): 1694-1704.

[6]Bernhoft A, Torp M, Clasen P E, Loes A K, Kristoffersen A B. Influence of agronomic and climatic factors on Fusarium infestation and mycotoxin contamination of cereals in Norway. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(7): 1129-1140.

[7]Zhang H, Van der Lee T, Waalwijk C, Chen W, Xu J, Xu J S, Zhang Y, Feng J. Population analysis of the Fusarium graminearum species complex from wheat in China show a shift to more aggressive isolates. PLoS One, 2012, 7(2): e31722.

[8]Zhang X, Zhou M, Ren L, Bai G, Ma H, Scholten O E, Guo P G, Lu W Z. Molecular characterization of Fusarium head blight resistance from wheat variety Wangshuibai. Euphytica, 2004, 139(1): 59-64.

[9]张旭, 邢锦城, 马鸿翔, 温云平, 陆维忠, 袁生. 江淮流域小麦赤霉病菌的遗传多样性. 江西农业大学学报, 2010, 32(6): 1146-1151.

Zhang X, Xing J C, Ma H X, Wen Y P, Lu W Z, Yuan S. Genetic diversity of Fusarium spp. isolates in Yangtze-huaihe river valley. Acta Agriculturae Universitis Jiangxiensis, 2010, 32(6): 1146-1151. (in Chinese)

[10]马鸿翔, 陆维忠. 小麦赤霉病抗性改良研究进展. 江苏农业学报, 2010, 26(1): 197-203.

Ma H X, Lu W Z. Progress on genetic improvement for resistance to Fusarium head blight in wheat. Jiangsu Journal of Agricultural Sciences, 2010, 26(1): 197-203. (in Chinese)

[11]Piekkola S, Turner P C, Abdel-Hamid M, Ezzat S, El-Daly M, El-Kafrawy S, Savchenko E, Poussa T, Woo J C S, Mykkanen H, El-Nezami H. Characterisation of aflatoxin and deoxynivalenol exposure among pregnant Egyptian women. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(6): 962-971.

[12]Ruiz M J, Franzova P, Juan-Garcia A, Font G. Toxicological interactions between the mycotoxins beauvericin, deoxynivalenol and T-2 toxin in CHO-K1 cells in vitro. Toxicon, 2011, 58(4): 315-326.

[13]Barros G, Alaniz Zanon M S, Palazzini J M, Haidukowski M, Pascale M, Chulze S. Trichothecenes and zearalenone production by Fusarium equiseti and Fusarium semitectum species isolated from Argentinean soybean. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(9): 1436-1442.

[14]Almeida M I, Almeida N G, Carvalho K L, Goncalves G A, Silva C N, Santos E A, Garcia J C, Vargas E A. Co-occurrence of aflatoxins B1, B2, G1 and G2, ochratoxin A, zearalenone, deoxynivalenol, and citreoviridin in rice in Brazil. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2012, 29(4): 694-703.

[15]Zhang Y, Caupert J. Survey of mycotoxins in U.S. distiller's dried grains with solubles from 2009 to 2011. Journal of Agricultural and Food Chemistry, 2012, 60(2): 539-543.

[16]Awad W A, Ghareeb K, Bohm J, Zentek J. Decontamination and detoxification strategies for the Fusarium mycotoxin deoxynivalenol in animal feed and the effectiveness of microbial biodegradation. Food Additives & Contaminants: Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment, 2010, 27(4): 510-520.

[17]Bejaoui H, Mathieu F, Taillandier P, Lebrihi A. Biodegradation of ochratoxin A by Aspergillus section Nigri species isolated from French grapes: a potential means of ochratoxin A decontamination in grape juices and musts. FEMS Microbiology Letters, 2006, 255(2): 203-208.

[18]Seeling K, Boguhn J, Strobel E, Danicke S, Valenta H, Ueberschar K H, Rodehutscord M. On the effects of Fusarium toxin contaminated wheat and wheat chaff on nutrient utilisation and turnover of deoxynivalenol and zearalenone in vitro (Rusitec). Toxicology in Vitro, 2006, 20(5): 703-711.

[19]Fuchs E, Binder E M, Heidler D, Krska R. Structural characterization of metabolites after the microbial degradation of type A trichothecenes by the bacterial strain BBSH 797. Food Additives and Contaminants, 2002, 19(4): 379-386.

[20]Shima J, Takase S, Takahashi Y, Iwai Y, Fujimoto H, Yamazaki M, Ochi K. Novel detoxification of the trichothecene mycotoxin deoxynivalenol by a soil bacterium isolated by enrichment culture. Applied and Environmental Microbiology, 1997, 63(10): 3825-3830.

[21]He C, Fan Y, Liu G, Zhang H. Isolation and identification of a strain of Aspergillus tubingensis with deoxynivalenol biotransformation capability. International Journal of Molecular Sciences, 2008, 9(12): 2366-2375.

[22]Ikunaga Y, Sato I, Grond S, Numaziri N, Yoshida S, Yamaya H, Hiradate S, Hasegawa M, Toshima H, Koitabashi M, Ito M, Karlovsky P, Tsushima S. Nocardioides sp. strain WSN05-2, isolated from a wheat field, degrades deoxynivalenol, producing the novel intermediate 3-epi-deoxynivalenol. Applied Microbiology and Biotechnology, 2011, 89(2): 419-427.

[23]Sato I, Ito M, Ishizaka M, Ikunaga Y, Sato Y, Yoshida S, Koitabashi M, Tsushima S. Thirteen novel deoxynivalenol-degrading bacteria are classified within two genera with distinct degradation mechanisms. FEMS Microbiology Letters, 2012, 327(2): 110-117.

[24]徐剑宏, 祭芳, 王宏杰, 王建伟, 林凡云, 史建荣. 脱氧雪腐镰刀菌烯醇降解菌的分离和鉴定. 中国农业科学, 2010, 43(22): 4635-4641.

Xu J H, Ji F, Wang H J, Wang J W, Lin F Y, Shi J R. Isolation and identification of deoxynivalenol degradation strains. Scientia Agricultura Sinica, 2010, 43(22): 4635-4641. (in Chinese)

[25]张忠辉, 洪青, 张国顺, 徐剑宏, 李顺鹏. 杀螟硫磷降解菌FDS-1的分离鉴定及其降解特性. 中国环境科学, 2005, 25(1): 52-56.

Zhang Z H, Hong Q, Zhang G S, Xu J H, Li S P. Studied on isolation identification of fenitrothion degradation bacteria FDS-1 and its degradation characteristics. China Environmental Science, 2005, 25(1): 52-56. (in Chinese)

[26]Azcona-Olivera J I, Ouyang Y L, Warner R L, Linz J E , Pestka J J. Effects of vomitoxin (deoxynivalenol) and cycloheximide on IL-2, 4, 5 and 6 secretion and mRNA levels in murine CD4+ cells. Food and Chemical Toxicology, 1995, 33(6): 433-441.

降解菌DDS-1产3-AC-DON氧化酶的酶学特性

Enzymatic Characteristics of 3-Acetyl Deoxynivalenol Oxidase by Devosia sp. DDS-1

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