首页 > 最新文献

微生物学报最新文献

英文 中文
[Gene cloning and biological function of CgRGS2 in Colletotrichum gloeosporioides]. [炭疽菌CgRGS2基因克隆及生物学功能研究]。
Pub Date : 2017-01-04
Manli Wu, Xiaoyu Li, Nan Zhang, Shuang Xu, Zhiqiang Liu

Objective: Regulators of G-protein signaling (RGS) are negative regulatory factors of G protein and play important roles in growth development and pathogenicity of plant pathogen. However, biological functions of RGS in Colletotrichum gloeosporioides have not been studied so far. We cloned an RGS gene of CgRGS2 in C. gloeosporioides and analyzed its biological function.

Methods: Gene CgRGS2 was cloned using PCR and analyzed. The gene-knockout mutant of CgRGS2 was obtained by homologous recombination, and the complementary strain was also built based on the mutant. Biological function of CgRGS2 was determined through phenotypic analysis.

Results: CgRGS2 encoded a 574-amino acids protein, containing an RGS function domain in the N terminal. Comparing to the wild type, the knockout mutant of CgRGS2 had slow growth, thick aerial hyphae, reduced conidia with multi-end germination, sensitive to oxidative stress and SDS, decreased pathogenicity.

Conclusion: Protein CgRGS2 was involved in regulation of vegetative growth, conidium production and germination, oxidative stress response, cell wall integrity and pathogenicity of C. gloeosporioides.

目的:G蛋白信号调控因子(regulatory of G-protein signaling, RGS)是G蛋白的负调控因子,在植物病原菌的生长发育和致病性中起重要作用。然而,RGS在炭疽菌中的生物学功能尚未得到研究。我们克隆了一种CgRGS2的RGS基因,并分析了其生物学功能。方法:采用PCR方法克隆CgRGS2基因并进行分析。通过同源重组获得CgRGS2基因敲除突变体,并以该突变体为基础构建互补菌株。通过表型分析确定CgRGS2的生物学功能。结果:CgRGS2编码了一个574个氨基酸的蛋白,在N端含有一个RGS功能域。与野生型相比,CgRGS2基因敲除突变体生长缓慢,气生菌丝粗大,分生孢子多端萌发减少,对氧化应激和SDS敏感,致病性降低。结论:CgRGS2蛋白参与了gloeosporioides营养生长、分生孢子产生和萌发、氧化应激反应、细胞壁完整性和致病性的调控。
{"title":"[Gene cloning and biological function of CgRGS2 in Colletotrichum gloeosporioides].","authors":"Manli Wu,&nbsp;Xiaoyu Li,&nbsp;Nan Zhang,&nbsp;Shuang Xu,&nbsp;Zhiqiang Liu","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>Regulators of G-protein signaling (RGS) are negative regulatory factors of G protein and play important roles in growth development and pathogenicity of plant pathogen. However, biological functions of RGS in Colletotrichum gloeosporioides have not been studied so far. We cloned an RGS gene of CgRGS2 in C. gloeosporioides and analyzed its biological function.</p><p><strong>Methods: </strong>Gene CgRGS2 was cloned using PCR and analyzed. The gene-knockout mutant of CgRGS2 was obtained by homologous recombination, and the complementary strain was also built based on the mutant. Biological function of CgRGS2 was determined through phenotypic analysis.</p><p><strong>Results: </strong>CgRGS2 encoded a 574-amino acids protein, containing an RGS function domain in the N terminal. Comparing to the wild type, the knockout mutant of CgRGS2 had slow growth, thick aerial hyphae, reduced conidia with multi-end germination, sensitive to oxidative stress and SDS, decreased pathogenicity.</p><p><strong>Conclusion: </strong>Protein CgRGS2 was involved in regulation of vegetative growth, conidium production and germination, oxidative stress response, cell wall integrity and pathogenicity of C. gloeosporioides.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Purification and characterization of endoglucanase Egn21 from Fusarium sp. Q7-31T]. [镰刀菌Q7-31T内切葡聚糖酶Egn21的纯化及特性研究]。
Pub Date : 2017-01-04
Xinyuan Chang, Zhanling Xie, Fengmei Zhang, Jieqiong Lei, Rongwei Cui, Shouyi Nie

Objective: The objective of this research was to study plant cell wall degradation enzymes from Fusarium sp. Q7-31T.

Methods: Strain was cultured in liquid medium with 1% (W/V) peptone as nitrogen source, 0.5% (W/V) oat straw as carbon source, 120 r/min shaking at 20 °C for 3 days. The endoglucanase Egn21 was purified by using Sephacry S-100 chromatography and DEAE-sepharose ion-exchange column chromatography. Then the enzymatic properties and MADIL-TOF-TOF identification were analyzed.

Results: The molecular weight and isoelectric point (pI) of Egn21 was 44.25 kDa and 4.91, respectively. Egn21 had optimal activity with carboxymethyl cellulose at 40 °C and pH 6.0, stable at 45 °C and pH between 5.0 and 8.0, inhibited by Fe2+, Ca2+, K+, Na+, Mn2+ and inactivated by Hg2+, whereas Co2+, Zn2+ and Mg2+ had no effect.

Conclusion: The enzymatic properties and MADIL-TOF-TOF results suggested that Egn21 belongs to GH5 family.

目的:对镰刀菌Q7-31T细胞壁降解酶进行研究。方法:以1% (W/V)蛋白胨为氮源,0.5% (W/V)燕麦秸秆为碳源,120 r/min, 20℃振荡培养3 d。采用Sephacry S-100层析和DEAE-sepharose离子交换柱层析纯化内切葡聚糖酶Egn21。然后对酶学性质和MADIL-TOF-TOF鉴定进行了分析。结果:Egn21的分子量为44.25 kDa,等电点为4.91。Egn21在40℃、pH为6.0时对羧甲基纤维素的活性最优,在45℃、pH为5.0 ~ 8.0时活性稳定,可被Fe2+、Ca2+、K+、Na+、Mn2+抑制,可被Hg2+灭活,而Co2+、Zn2+和Mg2+对Egn21的活性无影响。结论:酶学性质和MADIL-TOF-TOF结果表明Egn21属于GH5家族。
{"title":"[Purification and characterization of endoglucanase Egn21 from Fusarium sp. Q7-31T].","authors":"Xinyuan Chang,&nbsp;Zhanling Xie,&nbsp;Fengmei Zhang,&nbsp;Jieqiong Lei,&nbsp;Rongwei Cui,&nbsp;Shouyi Nie","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>The objective of this research was to study plant cell wall degradation enzymes from Fusarium sp. Q7-31T.</p><p><strong>Methods: </strong>Strain was cultured in liquid medium with 1% (W/V) peptone as nitrogen source, 0.5% (W/V) oat straw as carbon source, 120 r/min shaking at 20 °C for 3 days. The endoglucanase Egn21 was purified by using Sephacry S-100 chromatography and DEAE-sepharose ion-exchange column chromatography. Then the enzymatic properties and MADIL-TOF-TOF identification were analyzed.</p><p><strong>Results: </strong>The molecular weight and isoelectric point (pI) of Egn21 was 44.25 kDa and 4.91, respectively. Egn21 had optimal activity with carboxymethyl cellulose at 40 °C and pH 6.0, stable at 45 °C and pH between 5.0 and 8.0, inhibited by Fe2+, Ca2+, K+, Na+, Mn2+ and inactivated by Hg2+, whereas Co2+, Zn2+ and Mg2+ had no effect.</p><p><strong>Conclusion: </strong>The enzymatic properties and MADIL-TOF-TOF results suggested that Egn21 belongs to GH5 family.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086662","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Mutant construction and characterization of hfq in Mesorhizobium huakuii 7653R]. [华奎中干酪根菌7653R hfq突变体的构建与鉴定]。
Pub Date : 2017-01-04
Chuncao Ma, Xuejuan Zhou, Fuli Xie, Youguo Li

Objective: We studied the functions and characteristics of hfq gene in Mesorhizobium huakuii 7653R in adverse environment and symbiotic with its host plant.

Methods: The hfq mutant of 7653R was constructed via homologous recombination with small cloned fragments on suicide plasmids pK19mob to insert target gene. We applied 7653RΔhfq to characterize stress tolerance and symbiosis with host plant, in comparison with the complementary strains 7653R △hfq-C and the wild type.

Results: Mutant 7653RΔhfq presented lower growth rate, and higher mortality after heat shock-pretreated than that of the wild type, as well as the decreasing adaptability under the stress of 4.5% ethanol and 50 mmol H2O2. The defection of hfq affected the expression of some sRNAs in 7653R. Moreover, the mutant displayed significant reduced nodulation ability and nitrogenase activity compared with the wild type.

Conclusion: As a crucial post transcriptional regulatory factor, hfq plays an important role in Mesorhizobium Huakuii 7653R on both processes of stress resistance and symbiosis with the host plant Astragalus sinicus L.

目的:研究华葵中根菌7653R在逆境环境下及其与寄主植物共生时hfq基因的功能和特性。方法:在自杀质粒pK19mob上同源重组克隆小片段,构建7653R hfq突变体,插入目的基因。利用7653RΔhfq与互补菌株7653R△hfq-C和野生型进行比较,分析菌株的耐受性和与寄主植物的共生关系。结果:突变体7653RΔhfq经热休克预处理后的生长率低于野生型,死亡率高于野生型,在4.5%乙醇和50 mmol H2O2胁迫下的适应性下降。hfq的缺失影响了7653R中部分srna的表达。此外,与野生型相比,突变体的结瘤能力和氮酶活性显著降低。结论:hfq作为一个重要的转录后调控因子,在华葵中根菌7653R的抗逆性和与寄主植物黄芪的共生过程中都起着重要的作用。
{"title":"[Mutant construction and characterization of hfq in Mesorhizobium huakuii 7653R].","authors":"Chuncao Ma,&nbsp;Xuejuan Zhou,&nbsp;Fuli Xie,&nbsp;Youguo Li","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>We studied the functions and characteristics of hfq gene in Mesorhizobium huakuii 7653R in adverse environment and symbiotic with its host plant.</p><p><strong>Methods: </strong>The hfq mutant of 7653R was constructed via homologous recombination with small cloned fragments on suicide plasmids pK19mob to insert target gene. We applied 7653RΔhfq to characterize stress tolerance and symbiosis with host plant, in comparison with the complementary strains 7653R △hfq-C and the wild type.</p><p><strong>Results: </strong>Mutant 7653RΔhfq presented lower growth rate, and higher mortality after heat shock-pretreated than that of the wild type, as well as the decreasing adaptability under the stress of 4.5% ethanol and 50 mmol H2O2. The defection of hfq affected the expression of some sRNAs in 7653R. Moreover, the mutant displayed significant reduced nodulation ability and nitrogenase activity compared with the wild type.</p><p><strong>Conclusion: </strong>As a crucial post transcriptional regulatory factor, hfq plays an important role in Mesorhizobium Huakuii 7653R on both processes of stress resistance and symbiosis with the host plant Astragalus sinicus L.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Interaction between microflora and nitrogen nutrients in large intestine and its impacts on host health]. [大肠菌群与氮营养物质的相互作用及其对宿主健康的影响]。
Pub Date : 2017-01-04
Zhuang Liu, Junshi Shen, Weiyun Zhu

Proteins are not only the main building blocks for the construction of tissue, but also crucial for metabolic activity in animals. The microbial community colonized in the gastrointestinal tract plays an important role in host nutrients metabolism, especially nitrogen nutrients. Bacteria in small intestine could metabolize parts of amino acids (AAs), which further affects the systemic AAs metabolism of host. Compared with that in small intestine, the density of bacteria is much higher and the retention time of chyme is much longer in large intestine. On the one hand, the metabolism and community structure of microflora could be affected by nitrogen nutrients entering the large intestine. Thus, the metabolism of nitrogen nutrients by large intestinal microflora can lead to the formation of several metabolites, which are generally presumed to be detrimental for the host health. This review summarized the effects of dietary protein on the community structure of large intestinal microflora, the microbial metabolites of AAs in large intestine and their impacts on intestinal physiology and host health.

蛋白质不仅是组织结构的主要组成部分,而且对动物的代谢活动也至关重要。胃肠道中定植的微生物群落在宿主营养物质尤其是氮营养物质代谢中起着重要作用。小肠细菌能够代谢部分氨基酸,进而影响宿主机体对氨基酸的代谢。与小肠相比,大肠的细菌密度要高得多,食糜的滞留时间也要长得多。一方面,氮营养物质进入大肠会影响微生物群的代谢和群落结构。因此,大肠菌群对氮营养物质的代谢可导致几种代谢物的形成,这些代谢物通常被认为对宿主健康有害。本文就饲粮蛋白质对大肠菌群群落结构的影响、氨基酸在大肠内的微生物代谢产物及其对肠道生理和宿主健康的影响进行综述。
{"title":"[Interaction between microflora and nitrogen nutrients in large intestine and its impacts on host health].","authors":"Zhuang Liu,&nbsp;Junshi Shen,&nbsp;Weiyun Zhu","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Proteins are not only the main building blocks for the construction of tissue, but also crucial for metabolic activity in animals. The microbial community colonized in the gastrointestinal tract plays an important role in host nutrients metabolism, especially nitrogen nutrients. Bacteria in small intestine could metabolize parts of amino acids (AAs), which further affects the systemic AAs metabolism of host. Compared with that in small intestine, the density of bacteria is much higher and the retention time of chyme is much longer in large intestine. On the one hand, the metabolism and community structure of microflora could be affected by nitrogen nutrients entering the large intestine. Thus, the metabolism of nitrogen nutrients by large intestinal microflora can lead to the formation of several metabolites, which are generally presumed to be detrimental for the host health. This review summarized the effects of dietary protein on the community structure of large intestinal microflora, the microbial metabolites of AAs in large intestine and their impacts on intestinal physiology and host health.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36088294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[The nuclear import of Newcastle disease virus matrix protein depends on KPNB1 and Ran protein]. [新城疫病毒基质蛋白的核输入取决于KPNB1和Ran蛋白]。
Pub Date : 2017-01-04
Zhiqiang Duan, Xinqin Ji, Jouqiang Xu, Jiafu Zhao, Haixu Xu, Shunlin Hu, Xiufan Liu

Objective: The aim of this study was to identify the transport proteins that mediates the nuclear import of Newcastle disease virus (NDV) matrix (M) protein.

Methods: Chicken KPNA1 to KPNA6 gene and KPNB1 gene were cloned from DF-1 cells and then inserted into eukaryotic expression vectors. The constructed recombinant plasmids with a combination of grouping were transfected into HEK-293T cells to identify the transport proteins interacting with NDV M protein by co-immunoprecipitation (Co-IP) assay. Moreover, fluorescent co-localization assay was used to verify the transport proteins by co-expressing M and Ran protein mutant or M and its interactive protein deletant.

Results: The recombinant proteins could normally express in plasmid-transfected HEK-293T cells. Indirect immunofluorescence detection showed that the recombinant proteins except for Myc-KPNA2 displayed the same nuclear localization as NDV M protein. The results of Co-IP revealed that M protein could interact with KPNA1 and KPNB1. Further fluorescent co-localization indicated that co-expression of M and DN-KPNA1 did not change the nuclear localization of M, whereas co-expression of M and DN-KPNB1 or M and Ran-Q69L disrupted the nuclear localization of M, demonstrating that the nuclear import of M protein was dependent on KPNB1 and Ran protein.

Conclusion: KPNB1 and Ran protein jointly mediated the nuclear import of NDV M protein, showing that KPNB1 protein interacted with NDV M protein to form binary complex and then entered into the nucleus with the assistance of Ran protein.

目的:本研究旨在鉴定介导新城疫病毒(NDV)基质(M)蛋白核输入的转运蛋白。方法:从DF-1细胞中克隆鸡KPNA1 ~ KPNA6基因和KPNB1基因,并将其插入真核表达载体中。将组合分组构建的重组质粒转染HEK-293T细胞,用共免疫沉淀法(Co-IP)鉴定与NDV M蛋白相互作用的转运蛋白。此外,采用荧光共定位法通过共表达M和Ran蛋白突变体或M及其相互作用的蛋白缺失来验证转运蛋白。结果:重组蛋白能在转染HEK-293T细胞的质粒中正常表达。间接免疫荧光检测显示,除Myc-KPNA2外,重组蛋白与NDV M蛋白具有相同的核定位。Co-IP结果显示M蛋白可与KPNA1和KPNB1相互作用。进一步的荧光共定位表明,M与DN-KPNA1共表达不会改变M的核定位,而M与DN-KPNB1或M与Ran- q69l共表达会破坏M的核定位,说明M蛋白的核输入依赖于KPNB1和Ran蛋白。结论:KPNB1与Ran蛋白共同介导NDV M蛋白的核输入,表明KPNB1蛋白与NDV M蛋白相互作用形成二元复合物,并在Ran蛋白的协助下进入细胞核。
{"title":"[The nuclear import of Newcastle disease virus matrix protein depends on KPNB1 and Ran protein].","authors":"Zhiqiang Duan,&nbsp;Xinqin Ji,&nbsp;Jouqiang Xu,&nbsp;Jiafu Zhao,&nbsp;Haixu Xu,&nbsp;Shunlin Hu,&nbsp;Xiufan Liu","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>The aim of this study was to identify the transport proteins that mediates the nuclear import of Newcastle disease virus (NDV) matrix (M) protein.</p><p><strong>Methods: </strong>Chicken KPNA1 to KPNA6 gene and KPNB1 gene were cloned from DF-1 cells and then inserted into eukaryotic expression vectors. The constructed recombinant plasmids with a combination of grouping were transfected into HEK-293T cells to identify the transport proteins interacting with NDV M protein by co-immunoprecipitation (Co-IP) assay. Moreover, fluorescent co-localization assay was used to verify the transport proteins by co-expressing M and Ran protein mutant or M and its interactive protein deletant.</p><p><strong>Results: </strong>The recombinant proteins could normally express in plasmid-transfected HEK-293T cells. Indirect immunofluorescence detection showed that the recombinant proteins except for Myc-KPNA2 displayed the same nuclear localization as NDV M protein. The results of Co-IP revealed that M protein could interact with KPNA1 and KPNB1. Further fluorescent co-localization indicated that co-expression of M and DN-KPNA1 did not change the nuclear localization of M, whereas co-expression of M and DN-KPNB1 or M and Ran-Q69L disrupted the nuclear localization of M, demonstrating that the nuclear import of M protein was dependent on KPNB1 and Ran protein.</p><p><strong>Conclusion: </strong>KPNB1 and Ran protein jointly mediated the nuclear import of NDV M protein, showing that KPNB1 protein interacted with NDV M protein to form binary complex and then entered into the nucleus with the assistance of Ran protein.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Construction of recombinant Bacillus subtilis by co-expression of heterologous D-hydantoinase and N-carbamoylase]. [异种d -羟脲酶和n -氨基淀粉酶共表达构建重组枯草芽孢杆菌]。
Pub Date : 2017-01-04
Yameng Wang, Rui Ban, Lu Liu, Yu Shen

Objective: We aimed at co-expressing heterologous D-hydantoinase and N-carbamoylase in Bacillus subtilis, and evaluating the feasibility of producing D-p-hydroxyphenylglycine by the recombinant B. subtilis whole-cell catalysis.

Methods: The Paco expression cassette was combined with the coding sequence of hyd or sd1 gene as an artificial gene to express D-hydantoinase. The PAE expression cassette was combined with the coding sequence of adc gene as an artificial gene to express N-carbamoylase. The D-hydantoinase and N-carbamoylase co-expression plasmids pHCS(sd1+adc) and pHCY(hyd+adc) were constructed, using plasmid pHP13 as carrier; the co-expression plasmids pUCS(sd1+adc) was constructed, using plasmid pUB110 as carrier. The additional copy of acoR and sigL gene was integrated at chromosome. The skf and sdp gene were knocked out in B. subtilis. All recombinant strains bearing co-expression plasmid were characterized by analyzing whole-cell catalysis activity.

Results: In the recombinant strains with plasmid pHCY and with pHCS, the whole-cell catalytic activity reached 0.21 U/mL and 0.31 U/mL, respectively. After the over-expression of acoR, sigL, and high-copy-number pUCS, the whole-cell catalytic activity reached 1.0 U/mL.

Conclusion: Overexpression of acoR, sigL and the deletion of skf, sdp genes had significant effects on the catalysis activity of recombinant whole-cell.

目的:在枯草芽孢杆菌中共表达外源d -羟化酶和n -氨基甲酰化酶,评价重组枯草芽孢杆菌全细胞催化生产d -对羟基苯基甘氨酸的可行性。方法:将Paco表达盒与hyd或sd1基因编码序列结合,作为人工基因表达d -羟化酶。将PAE表达盒与adc基因编码序列结合,作为人工基因表达n -氨基甲酰酶。以质粒pHP13为载体,构建d -羟脲酶和n -氨基甲酰酶共表达质粒pHCS(sd1+adc)和pHCY(hyd+adc);以质粒pUB110为载体构建共表达质粒pUCS(sd1+adc)。acoR和sigL基因的附加拷贝整合在染色体上。skf和sdp基因在枯草芽孢杆菌中被敲除。所有携带共表达质粒的重组菌株通过全细胞催化活性分析进行了表征。结果:以pHCY和pHCS为质粒的重组菌株的全细胞催化活性分别达到0.21 U/mL和0.31 U/mL。过表达acoR、sigL和高拷贝数pUCS后,全细胞催化活性达到1.0 U/mL。结论:acoR、sigL的过表达和skf、sdp基因的缺失对重组全细胞的催化活性有显著影响。
{"title":"[Construction of recombinant Bacillus subtilis by co-expression of heterologous D-hydantoinase and N-carbamoylase].","authors":"Yameng Wang,&nbsp;Rui Ban,&nbsp;Lu Liu,&nbsp;Yu Shen","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>We aimed at co-expressing heterologous D-hydantoinase and N-carbamoylase in Bacillus subtilis, and evaluating the feasibility of producing D-p-hydroxyphenylglycine by the recombinant B. subtilis whole-cell catalysis.</p><p><strong>Methods: </strong>The Paco expression cassette was combined with the coding sequence of hyd or sd1 gene as an artificial gene to express D-hydantoinase. The PAE expression cassette was combined with the coding sequence of adc gene as an artificial gene to express N-carbamoylase. The D-hydantoinase and N-carbamoylase co-expression plasmids pHCS(sd1+adc) and pHCY(hyd+adc) were constructed, using plasmid pHP13 as carrier; the co-expression plasmids pUCS(sd1+adc) was constructed, using plasmid pUB110 as carrier. The additional copy of acoR and sigL gene was integrated at chromosome. The skf and sdp gene were knocked out in B. subtilis. All recombinant strains bearing co-expression plasmid were characterized by analyzing whole-cell catalysis activity.</p><p><strong>Results: </strong>In the recombinant strains with plasmid pHCY and with pHCS, the whole-cell catalytic activity reached 0.21 U/mL and 0.31 U/mL, respectively. After the over-expression of acoR, sigL, and high-copy-number pUCS, the whole-cell catalytic activity reached 1.0 U/mL.</p><p><strong>Conclusion: </strong>Overexpression of acoR, sigL and the deletion of skf, sdp genes had significant effects on the catalysis activity of recombinant whole-cell.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36087232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Heterologous expression, purification and characterization of phospholipase C from Bacillus cereus in Kluyveromyces lactis]. [蜡样芽孢杆菌磷脂酶C在乳酸克鲁维菌中的异源表达、纯化和特性研究]。
Pub Date : 2017-01-04
Chao Xiao, Liang Zhang, Yanyan Li, Yu Xin, Guoan Chen, Shengrong Yang

Objective: In this study, we constructed recombinant Kluyveromyces lactis strains to produce phospholipase C (PLC) of Bacillus cereus. The recombinant enzymes were purified and characterized.

Methods: We cloned the PLC encoding gene bcplc of Bacillus cereus. And the amplified fragments were inserted into pKLAC1 to obtain expression plasmids. K. lactis harboring the above plasmids was cultivated to express PLC that was purified by HisTrapTM affinity chromatography and characterized.

Results: PLC of B. cereus was cloned and expressed in K. lactis. The recombinant enzyme had shown activity of 19251 U/mg when using p-nitrophenyl phosphorycholine as substrate. Purified PLC exhibited optimum temperature at 80 °C and optimal pH at 9.0. The recombinant enzyme was stable below 40 °C and pH between 7.0 and 8.0. Cu2+ and Co2+ inhibited its activity whereas Zn2+, Mn2+, Ca2+ and Mg2+ stimulated its activity.

Conclusion: It is the first time to express and characterize the PLC gene in K. lactis. These research results provide reference for the study of recombinant PLC.

目的:构建重组乳酸克吕酵母菌生产蜡样芽孢杆菌磷脂酶C (PLC)的菌株。对重组酶进行了纯化和表征。方法克隆蜡样芽孢杆菌PLC编码基因bccp。将扩增片段插入pKLAC1中获得表达质粒。培养含有上述质粒的K. lactis表达PLC,通过HisTrapTM亲和层析纯化PLC并对其进行表征。结果:克隆出蜡样芽孢杆菌PLC,并在乳酸菌中表达。以对硝基苯基磷胆碱为底物时,酶活性为19251 U/mg。纯化后的PLC最适温度为80℃,最适pH为9.0。重组酶在40℃和7.0 ~ 8.0 pH范围内稳定。Cu2+和Co2+抑制其活性,而Zn2+、Mn2+、Ca2+和Mg2+则刺激其活性。结论:首次在乳酸菌中表达并鉴定了PLC基因。这些研究结果为重组PLC的研究提供了参考。
{"title":"[Heterologous expression, purification and characterization of phospholipase C from Bacillus cereus in Kluyveromyces lactis].","authors":"Chao Xiao,&nbsp;Liang Zhang,&nbsp;Yanyan Li,&nbsp;Yu Xin,&nbsp;Guoan Chen,&nbsp;Shengrong Yang","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>In this study, we constructed recombinant Kluyveromyces lactis strains to produce phospholipase C (PLC) of Bacillus cereus. The recombinant enzymes were purified and characterized.</p><p><strong>Methods: </strong>We cloned the PLC encoding gene bcplc of Bacillus cereus. And the amplified fragments were inserted into pKLAC1 to obtain expression plasmids. K. lactis harboring the above plasmids was cultivated to express PLC that was purified by HisTrapTM affinity chromatography and characterized.</p><p><strong>Results: </strong>PLC of B. cereus was cloned and expressed in K. lactis. The recombinant enzyme had shown activity of 19251 U/mg when using p-nitrophenyl phosphorycholine as substrate. Purified PLC exhibited optimum temperature at 80 °C and optimal pH at 9.0. The recombinant enzyme was stable below 40 °C and pH between 7.0 and 8.0. Cu2+ and Co2+ inhibited its activity whereas Zn2+, Mn2+, Ca2+ and Mg2+ stimulated its activity.</p><p><strong>Conclusion: </strong>It is the first time to express and characterize the PLC gene in K. lactis. These research results provide reference for the study of recombinant PLC.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Complete genome sequencing of polymalic acid-producing strain Aureobasidium pullulans CCTCC M2012223]. [产聚苹果酸菌株普鲁兰金黄色葡萄球菌CCTCC M2012223全基因组测序]。
Pub Date : 2017-01-04
Yongkang Wang, Xiaodan Song, Xiaorong Li, Sang-tian Yang, Xiang Zou

Objective: To explore the genome sequence of Aureobasidium pullulans CCTCC M2012223, analyze the key genes related to the biosynthesis of important metabolites, and provide genetic background for metabolic engineering.

Methods: Complete genome of A. pullulans CCTCC M2012223 was sequenced by Illumina HiSeq high throughput sequencing platform. Then, fragment assembly, gene prediction, functional annotation, and GO/COG cluster were analyzed in comparison with those of other five A. pullulans varieties.

Results: The complete genome sequence of A. pullulans CCTCC M2012223 was 30756831 bp with an average GC content of 47.49%, and 9452 genes were successfully predicted. Genome-wide analysis showed that A. pullulans CCTCC M2012223 had the biggest genome assembly size. Protein sequences involved in the pullulan and polymalic acid pathway were highly conservative in all of six A. pullulans varieties. Although both A. pullulans CCTCC M2012223 and A. pullulans var. melanogenum have a close affinity, some point mutation and inserts were occurred in protein sequences involved in melanin biosynthesis.

Conclusion: Genome information of A. pullulans CCTCC M2012223 was annotated and genes involved in melanin, pullulan and polymalic acid pathway were compared, which would provide a theoretical basis for genetic modification of metabolic pathway in A. pullulans.

目的:探索普鲁兰Aureobasidium pululans CCTCC M2012223的基因组序列,分析其重要代谢物生物合成相关的关键基因,为代谢工程提供遗传背景。方法:采用Illumina HiSeq高通量测序平台对芦兰CCTCC M2012223全基因组进行测序。然后,与其他5个普鲁兰品种进行片段组装、基因预测、功能注释和GO/COG聚类分析。结果:pululans CCTCC M2012223全基因组序列为30756831 bp,平均GC含量为47.49%,成功预测9452个基因。全基因组分析显示,普鲁兰芽孢杆菌CCTCC M2012223基因组组装大小最大。普鲁兰和聚苹果酸途径相关的蛋白序列在6个普鲁兰品种中均高度保守。虽然a . pululans CCTCC M2012223与a . pululans var. melanogenum亲缘关系密切,但在黑色素生物合成相关的蛋白序列中出现了一些点突变和插入。结论:对普鲁兰CCTCC M2012223基因组信息进行标注,并对黑色素、普鲁兰和聚苹果酸途径相关基因进行比较,为普鲁兰代谢途径的基因改造提供理论依据。
{"title":"[Complete genome sequencing of polymalic acid-producing strain Aureobasidium pullulans CCTCC M2012223].","authors":"Yongkang Wang,&nbsp;Xiaodan Song,&nbsp;Xiaorong Li,&nbsp;Sang-tian Yang,&nbsp;Xiang Zou","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Objective: </strong>To explore the genome sequence of Aureobasidium pullulans CCTCC M2012223, analyze the key genes related to the biosynthesis of important metabolites, and provide genetic background for metabolic engineering.</p><p><strong>Methods: </strong>Complete genome of A. pullulans CCTCC M2012223 was sequenced by Illumina HiSeq high throughput sequencing platform. Then, fragment assembly, gene prediction, functional annotation, and GO/COG cluster were analyzed in comparison with those of other five A. pullulans varieties.</p><p><strong>Results: </strong>The complete genome sequence of A. pullulans CCTCC M2012223 was 30756831 bp with an average GC content of 47.49%, and 9452 genes were successfully predicted. Genome-wide analysis showed that A. pullulans CCTCC M2012223 had the biggest genome assembly size. Protein sequences involved in the pullulan and polymalic acid pathway were highly conservative in all of six A. pullulans varieties. Although both A. pullulans CCTCC M2012223 and A. pullulans var. melanogenum have a close affinity, some point mutation and inserts were occurred in protein sequences involved in melanin biosynthesis.</p><p><strong>Conclusion: </strong>Genome information of A. pullulans CCTCC M2012223 was annotated and genes involved in melanin, pullulan and polymalic acid pathway were compared, which would provide a theoretical basis for genetic modification of metabolic pathway in A. pullulans.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Analysis of proposals received and funded in discipline of microbiology of the National Natural Science Foundation of China in 2016]. 【2016年国家自然科学基金微生物学科立项立项分析】。
Pub Date : 2017-01-04
Qiang Li, Weimin Li, Hongyan Shan, Guiqing Xiao, Mingzhang Wen, Quansheng Du

We summarized proposals submitted and funded in the discipline of microbiology of the Department of Life Sciences of National Natural Science Foundation of China in 2016. The traits and concerns in different sub-disciplines as well as distinctive funding programs were addressed, and the prior funding fields were prospected. The information may provide references for researchers who apply funding at the discipline of microbiology.

我们总结了2016年国家自然科学基金生命科学部微生物学科的申报和资助情况。分析了不同子学科的特点和关注点以及不同的资助项目,并展望了优先资助领域。这些信息可为申请微生物学科资助的研究人员提供参考。
{"title":"[Analysis of proposals received and funded in discipline of microbiology of the National Natural Science Foundation of China in 2016].","authors":"Qiang Li,&nbsp;Weimin Li,&nbsp;Hongyan Shan,&nbsp;Guiqing Xiao,&nbsp;Mingzhang Wen,&nbsp;Quansheng Du","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We summarized proposals submitted and funded in the discipline of microbiology of the Department of Life Sciences of National Natural Science Foundation of China in 2016. The traits and concerns in different sub-disciplines as well as distinctive funding programs were addressed, and the prior funding fields were prospected. The information may provide references for researchers who apply funding at the discipline of microbiology.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36088293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Effects of fatty acids and oxylipins on fungal growth, sporulation and aflatoxin production in Aspergillus]. 脂肪酸和氧化脂类对曲霉真菌生长、产孢和黄曲霉毒素产生的影响。
Pub Date : 2017-01-01
Shijuan Yan, Wenjie Huang, Chun-Ming Liu

Seeds with high oil contents are more susceptible to aflatoxin contamination after infected by Aspergillus species. However, in vitro studies showed that different types of fatty acids have striking difference on fungal growth, sporulation and aflatoxin biosynthesis in Aspergillus. Recent studies revealed that, although all fatty acids examined promote aflatoxin production, oxidized polyunsaturated fatty acids inhibit aflatoxin biosynthesis. The inhibiting effect is derived from oxylipins produced during autoxidation. In this article, we provide an overview for recent progress in fatty acids and oxylipins on fungal growth, sporulation and aflatoxin production in Aspergillus species.

含油量高的种子被曲霉侵染后,更容易受到黄曲霉毒素的污染。然而,体外研究表明,不同类型的脂肪酸对曲霉的真菌生长、产孢和黄曲霉毒素的生物合成有显著差异。最近的研究表明,虽然所有脂肪酸都促进黄曲霉毒素的产生,但氧化的多不饱和脂肪酸抑制黄曲霉毒素的生物合成。抑制作用来源于自氧化过程中产生的氧脂素。本文就脂肪酸和氧化脂类对曲霉真菌生长、产孢和黄曲霉毒素产生的影响的研究进展作一综述。
{"title":"[Effects of fatty acids and oxylipins on fungal growth, sporulation and aflatoxin production in Aspergillus].","authors":"Shijuan Yan,&nbsp;Wenjie Huang,&nbsp;Chun-Ming Liu","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Seeds with high oil contents are more susceptible to aflatoxin contamination after infected by Aspergillus species. However, in vitro studies showed that different types of fatty acids have striking difference on fungal growth, sporulation and aflatoxin biosynthesis in Aspergillus. Recent studies revealed that, although all fatty acids examined promote aflatoxin production, oxidized polyunsaturated fatty acids inhibit aflatoxin biosynthesis. The inhibiting effect is derived from oxylipins produced during autoxidation. In this article, we provide an overview for recent progress in fatty acids and oxylipins on fungal growth, sporulation and aflatoxin production in Aspergillus species.</p>","PeriodicalId":7120,"journal":{"name":"微生物学报","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36086663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
微生物学报
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1