黄杆菌中浒苔多糖降解有关基因的异源表达及酶活测定
作者：
徐展;刘芳芳;徐玉雯;闫达中;晁红军;...
期刊：
武汉轻工大学学报 ,2024年43(03):59-65 ISSN：2095-7386
作者机构：
武汉轻工大学,生命科学与技术学院,武汉 430023;[徐展; 晁红军; 徐玉雯; 闫达中; 刘芳芳; 陈静] 武汉轻工大学,生命科学与技术学院
关键词：
浒苔多糖;黄杆菌;多糖降解;木聚糖酶
摘要：
浒苔绿潮爆发带来了一系列环境问题,也对浒苔资源的开发利用提出了新挑战.从生物酶法降解浒苔的主要成分浒苔多糖入手,筛选出能降解浒苔多糖的菌株,对其进行16S rRNA基因鉴定,同时对其中表达量上调的木聚糖酶基因(GH43)进行异源表达,利用紫外-可见分光光度计比色法进行酶活测定及酶学性质研究.结果表明筛选到的浒苔多糖降解菌为黄杆菌(Algibacter pacificus),克隆表达的木聚糖酶的最适温度为50 ℃,最适pH约为5,粗酶液酶活为65.71 U/mL.实验可为由浒苔引起的海洋污染治理问题提供理论依据,同时为该菌株及木聚糖酶的工业化利用提供了参考,有利于实现对浒苔的资源化利用.
语种：
中文
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2,4-二甲基苯酚降解菌的筛选和生长优化
作者：
潘舜;胡丹;吴菁;闫达中;晁红军
期刊：
武汉轻工大学学报 ,2024年43(01):17-27 ISSN：2095-7386
作者机构：
武汉轻工大学,生命科学与技术学院,武汉 430023;[胡丹; 潘舜; 闫达中; 晁红军; 吴菁] 武汉轻工大学
关键词：
4-二甲基苯酚;微生物降解;环境污染;响应面分析
摘要：
2,4-二甲基苯酚作为许多工农业产品生产的中间体在环境中大量积累,由于其难以自然降解,造成了一定的环境污染,是一种顽固性的环境污染物.本研究从市政污水处理厂的活性污泥中分离出了一株能够高效降解2,4-二甲基苯酚的菌株4302,它能在24 h内降解2 mmol/L的2,4-二甲基苯酚,并基于单因素实验结果进行响应面实验优化菌株4302生长条件,结果表明:预测菌株4302生长的最适条件为:温度31.7 ℃、pH 7.4、底物浓度0.9 mmol/L,接种后培养14 h,在此条件下,OD600在理论上可达到0.117.酶活测定表明,菌株4302具有原儿茶酸3,4-双加氧酶酶活,未检测到邻苯二酚 2,3-双加氧酶和龙胆酸1,2-双加氧酶的酶活,说明2,4-二甲基苯酚在4302菌株中的代谢途径可能是通过原儿茶酸开环代谢.此研究为2,4-二甲基苯酚的降解提供微生物储备及对污染环境的修复提供科学的理论参考.
语种：
中文
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苯甲腈降解菌Rhodococcus sp.BN02的筛选和鉴定
作者：
陈丛;刘言琛;冯建英;刁硕祺;晁红军;...
期刊：
武汉轻工大学学报 ,2024年43(03):31-40,52 ISSN：2095-7386
作者机构：
武汉轻工大学生命科学与技术学院,武汉 430023;[刘言琛; 冯建英; 刁硕祺; 晁红军; 吴菁; 陈丛; 闫达中] 武汉轻工大学生命科学与技术学院
关键词：
腈类化合物;苯甲腈;红球菌;降解
摘要：
本研究从武汉市东西湖区金银湖附近水体污泥中提取以苯甲腈为唯一碳氮源,筛选出一株可高效降解腈类化合物的菌株.经生理生化测定、16S rRNA基因序列测定、比对和系统发育树构建,确认该菌为红球菌属(Rhodococcus),并命名为Rhodococcus sp.BN02.利用单因素实验和响应面实验对Rhodococcus sp.BN02以苯甲腈为唯一碳氮源的生长特性进行研究,发现其在28 ℃、pH 7和800 mg/L苯甲腈的条件下生长最适宜,最大OD600值可达0.471,并在2 d内可将苯甲腈完全降解.此外,本研究还考察了 BN02对不同腈类化合物的降解能力,它在48 h内可完全降解500 mg/L苯甲腈,在4 d内可完全降解500 mg/L的乙腈、丙烯腈和琥珀腈,对500 mg/L的2-氰基吡啶培养7 d后降解率可达83.84％.本研究为消除环境中苯甲腈及其中间代谢有毒污染物以及进一步探究苯甲腈代谢机制提供理论依据.
语种：
中文
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假单胞菌NyZ12基因组的可塑多变特征
作者：
张慧玲;谢琪冲;叶林燕;梁潼;闫达中
期刊：
微生物学通报 ,2023年50(04):1525-1537 ISSN：0253-2654
作者机构：
[张慧玲; 谢琪冲; 叶林燕; 梁潼; 闫达中] 武汉轻工大学生命科学与技术学院
关键词：
假单胞菌NyZ12;突变体;基因组;基因组可塑性
摘要：
【背景】假单胞菌是广泛存在于土壤、水体环境的微生物，其中Pseudomonas plecoglossicida NyZ12是一株能够以环己胺为唯一碳源和氮源生长的革兰氏阴性菌，其基因组达到7.0Mb左右。【目的】研究假单胞菌NyZ12的基因组是否具有可塑性和多变特征。【方法】以环己胺为唯一碳源和氮源生长的P. plecoglossicida NyZ12为研究对象，以琥珀酸或者代谢中间产物环己酮为碳源连续传代让其自然发生突变，然后筛选在以环己胺为唯一碳源和氮源的无机盐培养基上不能生长的突变体。将获得的突变体进行全基因组测序，并与野生型假单胞菌NyZ12的全基因组进行比对。【结果】以琥珀酸和环己酮为碳源分别筛选到一株突变体T1和T2，测序比对后发现假单胞菌突变体T1、T2的基因组发生大量的缺失和突变。对基因丢失的原因进行了分析，丢失的2个大片段中存在大量的重复序列、转座酶、转座子和原噬菌体。【结论】假单胞菌NyZ12的基因组具有可塑多变的特征。其可能的机制为进一步揭示微生物的适应和进化提供了参考。
语种：
中文
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浒苔多糖降解菌的筛选及其碳源利用谱分析
作者：
黎芯怡;王静涵;甄莉;徐展;戴景程;...
期刊：
广西科技大学学报 ,2023年34(03):123-131 ISSN：2095-7335
作者机构：
武汉轻工大学 生命科学与技术学院,湖北 武汉 430023;[甄莉] 中广核环保产业有限公司,广东 深圳 518038;[王静涵; 黎芯怡; 陈静; 闫达中; 戴景程; 徐展] 武汉轻工大学
关键词：
浒苔;浒苔多糖;浒苔多糖降解菌;碳源利用谱
摘要：
浒苔中含有大量的浒苔多糖，分解、利用及转化浒苔多糖对藻类资源利用以及海洋碳循环具有重要意义。为筛选出能降解浒苔多糖的微生物，提高浒苔生物量的回收利用效率，本研究利用微生物分离鉴定以及基质测试技术从浒苔绿潮中分离鉴定了浮游及浒苔附着微生物，成功地从分离到的菌株中筛选出能降解浒苔多糖的菌株，并选取黄杆菌Algibacter lectus S7和交替单胞菌Alteromonas confluentis A1-6作为代表菌株进行碳源利用谱分析。结果发现：黄杆菌Algibacter lectus S7可以分解利用浒苔多糖、木聚糖、葡萄糖、半乳糖和蔗糖；交替单胞菌Alteromonas confluentis A1-6可以分解利用浒苔多糖、葡萄糖、半乳糖、乳糖、木聚糖、蔗糖、可溶性淀粉、麦芽糖和糊精。从生长曲线可见，两菌株均能高效分解利用浒苔多糖，并分别揭示了两菌株对碳源利用情况，为海洋微生物对浒苔多糖的分解代谢研究及工业上提高浒苔生物量回收利用效率提供菌种资源及碳源参考。
语种：
中文
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一株苯二甲酸同分异构体降解菌的筛选和鉴定
作者：
向杨;李莎;闫达中;晁红军
期刊：
武汉轻工大学学报 ,2022年41(06):1-6 ISSN：2095-7386
作者机构：
武汉轻工大学 生命科学与技术学院,武汉430023;[向杨; 闫达中; 李莎; 晁红军] 武汉轻工大学
关键词：
邻苯二甲酸;间苯二甲酸;对苯二甲酸;微生物降解;环境污染
摘要：
苯二甲酸因广泛用于塑料生产而在环境中大量存在,它们造成了一定的环境污染.从污泥中分离出了一株能分别以三种苯二甲酸同分异构体为唯一碳源和能源生长的降解菌CHJ04.经形态观察、生化鉴定及16 S rRNA序列分析,鉴定该菌株与嗜染料菌(Pigmentiphaga sp.)一致性99.65％,将其命名为Pigmentiphaga sp.CHJ04.研究了菌株CHJ04对邻苯二甲酸、间苯二甲酸和对苯二甲酸三种同分异构体的降解情况,并采用单因素试验研究了不同试验条件(温度、pH)对菌株CHJ04生长的影响.结果表明:菌株CHJ04能分别以邻苯二甲酸、间苯二甲酸和对苯二甲酸三种物质为唯一碳源生长并将其降解,最适宜pH为5～9,最适宜的温度为37℃左右.酶活测定表明,菌株CHJ04具有原儿茶酸3,4-双加氧酶酶活,未检测到邻苯二酚2,3-双加氧酶和龙胆酸1,2-双加氧酶的酶活,说明在CHJ04菌株中,邻苯二甲酸、间苯二甲酸和对苯二甲酸代谢途径的可能通过原儿茶酸开环途径代谢的.此研究为苯二甲酸相关塑料环境污染的的生物修复提供了一定的理论依据.
语种：
中文
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酶联免疫吸附测定法检测淡水鱼中孔雀石绿残留
作者：
王洲;王君炎;闫达中;晁红军;吴菁
期刊：
食品安全导刊 ,2022年(02):76-80 ISSN：1674-0270
作者机构：
[王洲; 王君炎; 闫达中; 晁红军; 吴菁] 武汉轻工大学生命科学与技术学院
关键词：
酶联免疫吸附测定法;孔雀石绿;淡水鱼
摘要：
目的:了解武汉市常青花园小区在售淡水鱼产品的孔雀石绿污染情况。方法:利用酶联免疫吸附测定法对常青花园周边地区的3个菜市场和3个超市所售卖的常见淡水鱼进行检测。结果:共检测组织样本39份,其中检测出孔雀石绿的样本有21份,孔雀石绿检出率为53.8%;超出国家标准的样本有13份,不合格率为33.3%。结论:该区域在售淡水鱼中孔雀石绿的残留情况比较突出。由于试验样本的来源难以进一步追溯,确定孔雀石绿违规添加的环节难度较大,亟需市场监管部门对市售淡水鱼产品从养殖、运输到售卖全链条建立起清晰可查的溯源体系,以便更好地保障市民的食品安全。
语种：
中文
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六氯丁二烯降解菌BJ-04的筛选鉴定及降解特性
作者：
梁俊龙;张慧玲;张维树;汪丰;晁红军;...
期刊：
环境科学与技术 ,2022年45(2):30-36 ISSN：1003-6504
作者机构：
[梁俊龙; 晁红军; 吴菁; 张维树; 张慧玲; 汪丰; 闫达中] 武汉轻工大学生命科学与技术学院
关键词：
六氯丁二烯;生物降解;16S rRNA测序;产物分析
摘要：
六氯丁二烯是一种新型的持久性有机污染物,利用微生物降解环境中六氯丁二烯是一种无二次污染、非常绿色高效的方法。该研究从武汉东西湖区马池桥附近污泥中采样,通过连续传代驯化,分离筛选到一株能够以六氯丁二烯为唯一碳源生长的菌株,具有高效降解六氯丁二烯(HCBD)能力。16S rRNA基因测序鉴定菌株;通过气相色谱法和气相色谱-质谱联用法来测定底物降解及产物生成情况,并对降解菌株的抗生素抗性和底物广谱性进行探究。结果表明,该菌的16S r RNA序列与根癌农杆菌(Agrobacterium tumefaciens)具有99%的一致性,构建系统发育树显示该菌株与Agrobacterium tumefaciens聚为一支,将其命名为Agrobacterium sp. BJ-04,降解菌在72 h内可将1 mmol/L的HCBD降解至0.1 mmol/L左右,但是使用氯离子电极法未检测到Cl-浓度变化,气相色谱-质谱联用法检测到可能的中间产物正丁醇生成,此外该菌对氨苄青霉素、氯霉素、卡那霉素、链霉素、庆大霉素和萘啶酸均具有抗生素抗性,能够利用正己烷、环己烷、苯、正丁醇为碳源进行生长。对菌株BJ-04系统研究为将来用于处理废水及土壤中污染物六氯丁二烯提供理论和技术支持,也为充分利用微生物代谢解决环境污染问题提供菌株资源和技术储备。
语种：
中文
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棉酚降解菌的筛选、鉴定及棉粕固态发酵效果研究
作者：
汪丰;张维树;崔颖;叶林燕;李梦托;...
期刊：
食品工业科技 ,2022年 ISSN：1002-0306
作者机构：
[李梦托; 周焕; 张维树; 崔颖; 汪丰; 曾维斯; 叶林燕; 闫达中] 武汉轻工大学生命科学与技术学院;[刘楠] 武汉中粮食品科技有限公司
关键词：
棉酚;棉粕;生物降解;鉴定;拉乌尔菌属;发酵
摘要：
目的：筛选分离出棉酚降解菌株，研究其生长降解特性和对棉粕中棉酚脱毒效果，为深入研究微生物降解棉酚的机制提供依据。方法：从山东棉花产地采集土壤样本，以醋酸棉酚为唯一碳源，稀释涂布法分离筛选出一株具有棉酚降解能力的菌株，通过 16S rRNA 基因序列系统发育分析以及生理生化特征对菌株进行初步鉴定，确定菌属。用高效液相色谱测定其对棉酚的降解和棉粕固态发酵脱毒效果。结果：筛选到一株可以降解棉酚的细菌，命名为YL01，初步鉴定为拉乌尔菌属Raoultella sp.。该菌株除利用棉酚外，还可以邻苯二酚、原儿茶酸和对苯醌为唯一碳源生长。YL01在以棉酚为唯一碳源的无机盐液体培养基中生长10 d后，棉酚的降解率为48%；接种该菌株到棉粕中，通过固态发酵，8 d内棉粕中棉酚含量下降43%，蛋白含量增加10%。结论：本研究分离筛选到一株棉酚降解菌Raoultella sp. YL01，为进一步深入研究棉酚的微生物降解机制奠定了基础，其对运用生物处理技术消除棉粕中棉酚具有潜在的应用前景。
语种：
中文
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红球菌35R1中邻苯二酚1,2-双加氧酶的表达及纯化
作者：
何航航;姚霓红;吴菁;闫达中;晁红军
期刊：
武汉轻工大学学报 ,2021年40(03):20-23+72 ISSN：2095-7386
作者机构：
[姚霓红; 闫达中; 何航航; 晁红军; 吴菁] 武汉轻工大学生命科学与技术学院,武汉430023
关键词：
3,5-二甲基苯酚;邻苯二酚;邻苯二酚1,2-双加氧酶;基因克隆;异源表达
摘要：
3,5-二甲基苯酚(3,5-DMP)是一种重要的精细化工原料以及有机合成中间体,在工农业生产领域广泛应用并大量排放到环境中,对人类和环境造成了巨大的危害.Rhodococcus sp.35R1是一株以3,5-二甲基苯酚为唯一碳源和能源的高效降解菌.为验证菌株35R1中3,5-二甲基苯酚是通过邻苯二酚途径开环代谢的,通过生物信息学分析,寻找到35R1基因组上的邻苯二酚1,2-双加氧酶编码基因catA,通过一步克隆法将其连接到表达载体pET28a(+)上,然后,将其转化至表达宿主E.coli BL21(DE3)中,从而进行该基因的异源表达.结果成功在35R1基因组上扩增到catA基因,构建了重组质粒pET28a-catA,采用自诱导的方法诱导表达了邻苯二酚1,2-双加氧酶,纯化酶可催化邻苯二酚生成顺式粘糠酸.
语种：
中文
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Complete Genome Sequence of a Chlorobenzene Degrader, Pandoraea pnomenusa MCB032
作者：
Chao, Hong-Jun;Chen, Yuan-yuan;Wu, Jing;Yan, Da-Zhong* ;Zhou, Ning-Yi
期刊：
Current Microbiology ,2019年76(11):1235-1237 ISSN：0343-8651
通讯作者：
Yan, Da-Zhong
作者机构：
[Wu, Jing; Chao, Hong-Jun; Chen, Yuan-yuan; Yan, Da-Zhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.;[Chao, Hong-Jun; Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan, Hubei, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai, Peoples R China.
通讯机构：
[Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.
摘要：
Chlorobenzenes are ubiquitously distributed, highly persistent, and toxic environmental contaminants. Pandoraea pnomenusa MCB032 was isolated as a new dominant chlorobenzene-utilizing strain from a functionally stable bioreactor during the treatment of chlorobenzenes when strain Burkholderia sp. JS150 disappeared. In study, we report the complete genome sequence of strain MCB032 which consists of a circular chromosome and three plasmids, which are ~ 6Mb in length with 5450 open reading frames—12 encoding rRNAs and 77 encoding tRNAs. We further identified 17 putative genes encoding the enzymes involved in the methyl-accepting chemotaxis proteins in sensing chemical gradients during chemotaxis. The annotated complete genome sequence of this strain will provide genetic insights into the degradation of chlorinated aromatic compounds. The information will empower the elucidation of chlorobenzene affinity hierarchy and species succession in the bioreactor. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.
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Draft Genome Sequence of Cyclohexylamine-Degrading Strain Acinetobacter sp. YT-02 Isolated (vol 75, pg 284, 2018)
作者：
Yan, Da-Zhong* ;Gan, Ya-Ting;Zhou, Hui;Liu, Jun;Li, Xin
期刊：
Current Microbiology ,2019年76(9):1092-1092 ISSN：0343-8651
通讯作者：
Yan, Da-Zhong
作者机构：
[Zhou, Hui; Gan, Ya-Ting; Liu, Jun; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.
通讯机构：
[Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.
摘要：
The original version of this article unfortunately contained a mistake in the Fig. S1 of supplementary material. It is corrected with this erratum.
语种：
英文
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In Vitro Packaging Mediated One-Step Targeted Cloning of Natural Product Pathway
作者：
Tao, Weixin* ;Chen, Li;Zhao, Chunhua;Wu, Jing;Yang, Dazhong;...
期刊：
ACS Synthetic Biology ,2019年8(9):1991-1997 ISSN：2161-5063
通讯作者：
Tao, Weixin;Sun, Yuhui
作者机构：
[Sun, Yuhui; Tao, Weixin; Deng, Zixin; Chen, Li; Tao, WX; Sun, YH] Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan 430071, Hubei, Peoples R China.;[Sun, Yuhui; Tao, Weixin; Deng, Zixin; Chen, Li; Tao, WX; Sun, YH] Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China.;[Zhao, Chunhua] Huazhong Univ Sci & Technol, Tongji Med Coll, Sch Pharm, Wuhan 430030, Hubei, Peoples R China.;[Wu, Jing; Yang, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Hubei, Peoples R China.
通讯机构：
[Tao, WX; Sun, YH] M;[Tao, WX; Sun, YH] W;Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan 430071, Hubei, Peoples R China.;Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China.
关键词：
direct cloning;natural product;in vitro packaging;CRISPR/Cas9;gene editing
摘要：
Direct cloning of natural product pathways for efficient refactoring and heterologous expression has become an important strategy for microbial natural product research and discovery, especially for those kept silent or poorly expressed in the original strains. Accordingly, the development of convenient and efficient cloning approaches is becoming increasingly necessary. Here we presented an in vitro packaging mediated cloning approach that combines CRISPR/Cas9 system with in vitro λ packaging system, for targeted cloning of natural product pathways. In such a scheme, pathways of Tü3010 (27.4 kb) and sisomicin (40.7 kb) were respectively cloned, and stuR was further depicted to positively regulate Tü3010 production. In vitro packaging mediated approach not only enables to activate cryptic pathways, but also facilitates refactoring or interrogating the pathways in conjunction with various gene editing systems. This approach features an expedited, convenient, and generic manner, and it is conceivable that it may be widely adopted for targeted cloning of the natural product pathways. © 2019 American Chemical Society.
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The cyclase-associated protein ChCAP is important for regulation of hyphal growth, appressorial development, penetration, pathogenicity, conidiation, intracellular cAMP level, and stress tolerance in Colletotrichum higginsianum
作者：
Zhu, Wenjun;Xu, Xiaowen;Peng, Fang;Yan, Da-zhong;Zhang, Shaopeng;...
期刊：
Plant Science ,2019年283:1-10 ISSN：0168-9452
通讯作者：
Wei, Wei;Chen, Weidong
作者机构：
[Wu, Jing; Zhang, Shaopeng; Zhu, Wenjun; Peng, Fang; Yan, Da-zhong; Xu, Ran; Li, Xin] Wuhan Polytech Univ, Coll Biol & Pharmaceut Engn, Wuhan 430023, Hubei, Peoples R China.;[Xu, Xiaowen] Hubei Acad Forestry, Wuhan 430075, Hubei, Peoples R China.;[Wei, Wei; Chen, Weidong; Wei, W; Chen, WD] Washington State Univ, USDA ARS, Dept Plant Pathol, Pullman, WA 99164 USA.
通讯机构：
[Wei, W; Chen, WD] W;Washington State Univ, USDA ARS, Dept Plant Pathol, Pullman, WA 99164 USA.
关键词：
ChCAP;Colletotrichum higginsianum;Intracellular cAMP level;Pathogenicity;cAMP signaling pathway
摘要：
Colletotrichum higginsianum causes anthracnose disease in a wide range of cruciferous crops and has been used as a model system to study plant-pathogen interactions and pathogenicity of hemibiotrophic plant pathogens. Conidiation, hyphae growth, appressorial development and appressorial penetration are significant steps during the infection process of C. higginsianum. However, the mechanisms of these important steps during infection remain incompletely understood. To further investigate the mechanisms of the plant-C. higginsianum interactions during infection progress, we characterized Cyclase-Associated Protein (ChCAP) gene. Deletion of the ChCAP gene resulted in reduction in conidiation and hyphal growth rate. The pathogenicity of DeltaChCAP mutants was significantly reduced with much smaller lesion on the infected leaves compared to that of wild type strain with typically water-soaked and dark necrotic lesions on Arabidopsis leaves. Further study demonstrated that the appressorial formation rate, turgor pressure, penetration ability and switch from biotrophic to necrotrophic phases decreased obviously in DeltaChCAP mutants, indicating that the attenuated pathogenicity of DeltaChCAP mutants was due to these defective phenotypes. In addition, the DeltaChCAP mutants sectored on PDA with abnormal, dark color, vesicle-like colony morphology and hyphae tip. Moreover, the DeltaChCAP mutants had a reduced intracellular cAMP levels and exogenous cAMP can partially rescue the defects of DeltaChCAP mutants in appressorial formation and penetration rate, but not in colony morphology, conidial shape and virulence, indicating that ChCAP is a key component in cAMP signaling pathway and likely play other roles in biology of C. higginsianum. In summary, our findings support the role of ChCAP in regulating conidiation, intracellular cAMP level, hyphal growth, appressorial formation, penetration ability and pathogenicity of this hemibiotrophic fungus.
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Characterization of a New Cyclohexylamine Oxidase From Acinetobacter sp YT-02
作者：
Zhou, Hui;Han, Zheng-gang;Fang, Ti;Chen, Yuan-yuan;Ning, Shang-bo;...
期刊：
Frontiers in Microbiology ,2018年9(NOV):2848 ISSN：1664-302X
通讯作者：
Yan, Da-zhong
作者机构：
[Han, Zheng-gang; Chen, Yuan-yuan; Ning, Shang-bo; Yan, Da-zhong; Zhou, Hui; Gan, Ya-ting] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.;[Fang, Ti] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan, Hubei, Peoples R China.
通讯机构：
[Yan, Da-zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.
关键词：
Acinetobacter sp. YT-02;biodegradation;cyclohexanone;cyclohexylamine;cyclohexylamine oxidase
摘要：
Cyclohexylamine (CHAM) is widely used in various industries, but it is harmful to human beings and the environment. Acinetobacter sp. YT-02 can degrade CHAM via cyclohexanone as an intermediate. In this study, the cyclohexylamine oxidase (CHAO) gene from Acinetobacter sp. YT-02 was cloned. Amino acid sequence alignment indicated that the cyclohexylamine oxidase (CHAOYT-02) was 48% identical to its homolog from Brevibacterium oxydans IH-35A (CHAOIH-35). The enzyme was expressed in Escherichia coli BL21 (DE3), and purified to apparent homogeneity by Ni-affinity chromatography. The purified enzyme was proposed to be a dimer of molecular mass of approximately 91 kDa. The enzyme exhibited its maximum activity at 50°C and at pH 7.0. The enzyme was thermolabile as demonstrated by loss of important percentage of its maximal activity after 30 min incubation at 50 deg;C. Metal ions Mg2+, Co2+, and K+ had certain inhibitory effect on the enzyme activity. The kinetic parameters Km and Vmax were 0.25 ± 0.02 mM and 4.3 ± 0.083 μM min-1, respectively. The biochemical properties, substrate specificities, and three-dimensional structures of CHAOYT-02 and CHAOIH-35 were compared. Our results are helpful to elucidate the mechanism of microbial degradation of CHAM in the strain YT-02. In addition, CHAOYT-02, as a potential biocatalyst, is promising in controlling CHAM pollution and deracemization of chiral amines. © 2007 - 2018 Frontiers Media S.A.
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Draft Genome Sequence of Cyclohexylamine-Degrading Strain Acinetobacter sp YT-02 Isolated
作者：
Yan, Da-Zhong* ;Gan, Ya-Ting;Zhou, Hui;Liu, Jun;Li, Xin
期刊：
Current Microbiology ,2018年75(3):284-287 ISSN：0343-8651
通讯作者：
Yan, Da-Zhong
作者机构：
[Zhou, Hui; Gan, Ya-Ting; Liu, Jun; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.
通讯机构：
[Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, 68 Xuefu South Rd, Wuhan 430023, Hubei, Peoples R China.
摘要：
Acinetobacter sp. YT-02, a Gram-negative bacterium isolated from the activated sludge from a sodium N-cyclohexylsulfamate production plant, has the ability to degrade cyclohexylamine. It was classified as a member of Acinetobacter sp., a Gram-negative bacterium, sharing a 16S rRNA gene sequence identity of 99% with Acinetobacter guangdongensis strain 1NM-4. It could degrade 10 mmol/L cyclohexylamine within 22 h. Based on the identified metabolite, the metabolic pathway of cyclohexylamine could be postulated as it was degraded via cyclohexanone. Draft genome sequence of this strain (2,993, 647 bp of chromosome length) is presented here. We further identified the genes encoding the enzymes involved in cyclohexylamine oxidation to cyclohexanone and the subsequent downstream metabolic pathway of cyclohexanone oxidation. Strain YT-02 has the potentiality to be applied in the treatment of the pollutant cyclohexylamine, and it could also be treated as a research material to study the degradation mechanism of cyclohexylamine.
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英文
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Taraxerol Induces Cell Apoptosis through A Mitochondria-Mediated Pathway in HeLa Cells
作者：
Yao, Xiangyang;Lu, Binyu;Lu, Chaotian;Bai, Qin;Yan, Dazhong;...
期刊：
CELL JOURNAL ,2017年19(3):512-519 ISSN：2228-5806
通讯作者：
Xu, Hui
作者机构：
[Yao, Xiangyang; Bai, Qin; Xu, Hui; Lu, Chaotian] Bengbu Univ, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.;[Lu, Binyu] Fudan Univ, Sch Pharm, Shanghai, Peoples R China.;[Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Hubei, Peoples R China.
通讯机构：
[Xu, Hui] B;Bengbu Univ, Dept Biol & Food Engn, 1866 Caoshan Rd, Bengbu 233030, Peoples R China.
关键词：
Apoptosis;HeLa Cells;Mitochondria;Taraxerol
摘要：
Objective: Taraxerol acetate has potent anti-cancer effects via the induction of apoptosis, autophagy, cell cycle arrest, and inhibition of cell migration. However, whether taraxerol induced apoptosis and its underlying mechanisms of action is not clear. In the present study, we assess the effects of taraxerol on the mitochondrial apoptotic pathway and determine the release of cytochrome c to the cytosol and activation of caspases. Materials and Methods: In this experimental study, we mainly investigated the effect of taraxerol on HeLa cells. We tested cell viability by the MTT assay and morphologic changes, analyzed apoptosis by DAPI staining and flow cytometry. We also determined reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) using a Microplate Reader. In addition, the apoptotic proteins were tested by Western blot. Results: Taraxerol enhanced ROS levels and attenuated the MMP (Δψm) in HeLa cells. Taraxerol induced apoptosis mainly via the mitochondrial pathway including the release of cytochrome c to the cytosol and activation of caspases 9 and 3, and anti-poly (ADP-ribose) polymerase (PARP). Taraxerol could induce the down-regulation of the anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax. It suppressed the PI3K/ Akt signaling pathway. Conclusion: These results demonstrated that taraxerol induced cell apoptosis through a mitochondria-mediated pathway in HeLa cells. Thus, taraxerol might be a potential anti-cervical cancer candidate. Copyright © 2017 Powered By Niyasan Co.
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英文
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Solanesol induces the expression of heme oxygenase-1 via p38 and Akt and suppresses the production of proinflammatory cytokines in RAW264.7 cells
作者：
Yao, Xiangyang* ;Lu, Binyu;Lu, Chaotian;Bai, Qin;Yan, Dazhong;...
期刊：
FOOD & FUNCTION ,2017年8(1):132-141 ISSN：2042-6496
通讯作者：
Yao, Xiangyang
作者机构：
[Yao, Xiangyang; Bai, Qin; Xu, Hui; Lu, Chaotian; Wu, Yanli; Hong, Zibing] Bengbu Univ, Dept Biol & Food Engn, Bengbu, Peoples R China.;[Lu, Binyu] Fudan Univ, Sch Pharm, Shanghai, Peoples R China.;[Yan, Dazhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan, Peoples R China.
通讯机构：
[Yao, Xiangyang] B;Bengbu Univ, Dept Biol & Food Engn, Bengbu, Peoples R China.
摘要：
The aim of the present study was to examine the anti-inflammatory effect of solanesol and to elucidate the underlying mechanisms. Heme oxygenase-1 (HO-1) plays an important role in cytoprotection against oxidative stress and inflammation. Solanesol induced HO-1 expression both at the level of mRNA and proteins, resulting in increased HO-1 activity. Solanesol treatment enhanced the level of the phosphorylated form, nuclear translocation, ARE-binding, and transcriptional activity of Nrf2. p38 and Akt contributed to ARE-driven HO-1 expression. Solanesol activated both p38 and Akt, and treatments with SB203580 (a p38 kinase inhibitor), LY294002 (an Akt inhibitor), specific p38 siRNA and Akt siRNA suppressed the solanesol-induced activation of Nrf2, resulting in a decrease in HO-1 expression. Solanesol also elevated the autophagic protein LC3B-II level. SnPP (a HO-1 inhibitor) and HO-1 siRNA markedly abolished the anti-inflammatory effect of solanesol against LPS-induced cell damage. Likewise, SB203580, LY294002, 3-MA and Baf-A1 inhibited the solanesol-induced anti-inflammatory effect. These studies demonstrate that solanesol attenuates inflammation by HO-1 induction via p38 and Akt signaling. Thus, it is quite plausible that HO-1 induction by solanesol could trigger anti-inflammatory pathways including limiting LPS-stimulated cytokine production through autophagic signaling via p38 and Akt. © 2017 The Royal Society of Chemistry.
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英文
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Genome-wide identification and characterization of genes encoding cyclohexylamine degradation in a novel cyclohexylamine-degrading bacterial strain of Pseudomonas plecoglossicida NyZ12
作者：
Yan, Da-Zhong* ;Li, Xin;Li, Cun-Zhi;Mao, Ling-Qi;Chi, Xiang-Qun;...
期刊：
Journal of Biotechnology ,2017年251:166-173 ISSN：0168-1656
通讯作者：
Yan, Da-Zhong
作者机构：
[Li, Cun-Zhi; Mao, Ling-Qi; Yan, Da-Zhong; Li, Xin] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Chi, Xiang-Qun; Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China.;[Liu, Dong-Yan] Shanghai Normal Univ, Coll Life & Environm Sci, Shanghai 200234, Peoples R China.
通讯机构：
[Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.
关键词：
Bacteria;Carbon;Encoding (symbols);Genes;Metabolism;Polymerase chain reaction;RNA;Signal encoding;Transcription;Bioinformatics analysis;Catabolism;Cyclohexanone monooxygenase;Cyclohexanones;Cyclohexylamine;Gram-negative bacteria;Pseudomonas plecoglossicida;Reverse transcription PCR;Biodegradation;adipic acid;amine oxidase (flavin containing);cyclohexane;cyclohexanol;cyclohexanone;cyclohexylamine;cyclohexylamine oxidase;oxidoreductase;plasmid DNA;unclassified drug;cyclohexylamine derivative;Article;bacterial gene;bacterial genome;bacterium culture;carbon source;chaA gene;controlled study;gene cluster;gene identification;genome analysis;microbial degradation;nonhuman;nucleotide sequence;operon;priority journal;Pseudomonas;Pseudomonas plecoglossicida NyZ12;reverse transcription polymerase chain reaction;bacterial gene;bioremediation;genetics;metabolism;Pseudomonas;Biodegradation, Environmental;Cyclohexylamines;Genes, Bacterial;Genome, Bacterial;Oxidoreductases;Pseudomonas
摘要：
The Gram-negative strain of Pseudomonas plecoglossicida NyZ12 isolated from soil has the ability to degrade cyclohexylamine (CHAM). The genes encoding CHAM degradation by gram-negative bacteria, however, have not been reported previously. In this study, ORFs predicted to encode CHAM degradation by NyZ12 were identified by bioinformatics analysis. Differential expression of the proposed ORFs was analyzed via RNA-seq and quantitative reverse transcription-PCR (qRT-PCR), using RNA extracted from NyZ12 cultured with or without CHAM addition. One CHAM-inducible ORF, RK21_02867 predicted to encode a cyclohexanone monooxygenase (ChnB) was disrupted, as were five ORFs, RK21_00425, RK21_02631, RK21_04207, RK21_04637 and RK21_05539, that had weak homology to the only known cyclohexylamine oxidase (CHAO encoded by chaA) found in Brevibacterium oxydans IH-35A. We also found that a tandem array of five ORFs (RK21_02866-02870) shared homology with those in an operon responsible for oxidation of cyclohexanone to adipic acid, although the ORFs in strain NyZ12 were arranged in a different order with previously found in cyclohexane, cyclohexanol or cyclohexanone degradation strains. The ORFs in this cluster were all up-regulated when CHAM was supplied as the sole carbon source. When one of these five genes, RK21_02867 encoding cyclohexanone (CHnone) monooxygenase, was knocked out, NyZ12 could not grow on CHAM, but it accumulated equimolar amounts of CHnone. Our results show that strain NyZ12 metabolized CHAM directly to CHnone which was then further metabolized to adipate. Despite clearly identifying genes encoding the steps for metabolism of CHAM metabolites, not every one of the putative chaAs was differentially expressed in the presence of CHAM and deletion of each one individually did not completely eliminate the capacity of NyZ12 to degrade CHAM, though it did reduce its growth in several instances. Our results suggest that there is genetic redundancy encoding the initial step in the oxidation of CHAM to CHnone in NyZ12 and that its CHAOs differ considerably from the ChaA, originally described in Brevibacterium oxydans IH-35A. © 2017 Elsevier B.V.
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Complete genome sequence of the cyclohexylamine-degrading Pseudomonas plecoglossicida NyZ12
作者：
Li, Xin;Li, Cun-Zhi;Mao, Ling-Qi;Yan, Da-Zhong* ;Zhou, Ning-Yi
期刊：
Journal of Biotechnology ,2015年199:29-30 ISSN：0168-1656
通讯作者：
Yan, Da-Zhong
作者机构：
[Li, Cun-Zhi; Mao, Ling-Qi; Li, Xin; Yan, Da-Zhong] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China.;[Zhou, Ning-Yi] Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.
通讯机构：
[Yan, Da-Zhong] W;Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, Wuhan 430023, Peoples R China.
关键词：
Bacteria;Genetic engineering;Oxidation;Complete genomes;Cyclohexanones;Cyclohexylamine;Gene clusters;Genome sequences;Gram-negative bacteria;Metabolic pathways;Pseudomonas plecoglossicida;Genes;adipic acid;cyclohexylamine;ribosome RNA;transfer RNA;cyclohexylamine derivative;Article;bacterial chromosome;bacterial genome;bacterial strain;biodegradation;gene cluster;gene sequence;Gram negative bacterium;nonhuman;nucleotide sequence;oxidation;priority journal;Pseudomonas;Pseudomonas plecoglossicida NyZ12;soil microflora;bacterial genome;genetics;metabolism;molecular genetics;multigene family;Pseudomonas;Negibacteria;Pseudomonas plecoglossicida;Cyclohexylamines;Genome, Bacterial;Molecular Sequence Data;Multigene Family;Pseudomonas
摘要：
Pseudomonas plecoglossicida NyZ12 (CCTCC AB 2015057), a Gram-negative bacterium isolated from soil, has the ability to degrade cyclohexylamine. The complete genome sequence of this strain (6,233,254. bp of chromosome length) is presented, with information about the genes of characteristic enzymes responsible for cyclohexylamine oxidation to cyclohexanone and the integrated gene cluster for the metabolic pathway of cyclohexanone oxidation to adipate. © 2015 Elsevier B.V.
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