[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chrom...[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chromatography and fine chromatography. [Result]The optimized separation and purification conditions were that： Supernatant was treated by trichloroacetic acid, then by ultrafiltration desalt and anion exchange chromatography. Strong anion Q F. F. was better than weak anion DEAE F.F. The proper balanced solution was Tris-HCI （ pH 8.0）. The proper conductivity was 6 ms/cm. The maximum applied sample was 240 ATU/ml to matrix of strong anion Q F. F. This optimized procedure was magnified in strong anion exchange HiPrep 16/10Q with the 90% recovery and 70.2% purity. The purification of gel filtration of Sephacryl S-100 to hirudin was not relative to flow rate within certain scope. The application size of sample was 10 ml. The purity checked by HPLC was 95.1%, and the recovery was 93%, and the band of SDS-PAGE was single. [ Conclusion] The research provided the reference of the further industrialization separation and purification of hiruin.展开更多
[Objectives] This paper aims to explore the possibility to intercrop garlic with pomegranate tree to control pomegranate wilt.[Methods] Root exudates of garlic is cultivated in 1/5 concentration of MS solution and dis...[Objectives] This paper aims to explore the possibility to intercrop garlic with pomegranate tree to control pomegranate wilt.[Methods] Root exudates of garlic is cultivated in 1/5 concentration of MS solution and distilled water is examined in lab to test their effect to growth of mycelia of pomegranate wilt pathogen（Ceratocystis fimbriata）and multiplication of Bacillus subtilis.[Results] The result shows that garlic root exudates whatever cultivated in MS solution or distilled water could not inhibit or promote mycelia growth of C.fimbriata.However,garlic root exudates cultivated in both methods effectively promote multiplication of B.subtilis.[Conclusions] It is suggested that intercropping garlic with pomegranate tree by combining application B.subtilis could be a promising way to prevent pomegranate wilt spread in practice.展开更多
Two plastid division genes, NtFtsZ1 and NtFtsZ2 isolated from Nicotiana tabacum L. were fused with gfp and expressed in Escherichia coli . The regular localizations of full length NtFtsZs∶GFP along the fil...Two plastid division genes, NtFtsZ1 and NtFtsZ2 isolated from Nicotiana tabacum L. were fused with gfp and expressed in Escherichia coli . The regular localizations of full length NtFtsZs∶GFP along the filamentous bacteria indicated that the NtFtsZs could recognize the potential division sites in E. coli and be polymerized with heterogeneous FtsZ from bacteria. The overexpression of NtFtsZs ∶ gfp inhibited the division of host strain cells and resulted in the long filamentous bacterial morphology. These results suggested that eukaryotic ftsZs have similar function to their prokaryotic homologs. Meanwhile, the different deletions of motifs of NtFtsZs are also employed to investigate the functions of these proteins in E. coli . The results showed that the C_terminal domains of NtFtsZs were related to the correct localization of NtFtsZs in E. coli and the N_terminal domains of NtFtsZs were responsible for the polymerization of homogeneous and heterogeneous FtsZ proteins. The significance of these results in understanding the functions of NtFtsZs in plastid division were discussed.展开更多
In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY- 3, which produced a bacteriocin-like substance （BLS）, was characterized to be a strain of Baci...In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY- 3, which produced a bacteriocin-like substance （BLS）, was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16S rDNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria （including Staphylococcus aureus and B. subtillis） and some fimgi （including Penicillium glaucum, Aspergillus niger and Aspergillus flavus）. The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100~C for 60 min without losing any activity and stable over a wide pH range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid （HC1） and gel filteration （Sephadex G-100）. SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 kDa. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.展开更多
The vast majority of herbaceous plants engage into arbuscular mycorrhizal (AM) symbioses and consideration of their mycorrhizal status should be embodied in studies of plant-microbe interactions. To establish reliable...The vast majority of herbaceous plants engage into arbuscular mycorrhizal (AM) symbioses and consideration of their mycorrhizal status should be embodied in studies of plant-microbe interactions. To establish reliable AM contrasts, however, a sterilized re-inoculation procedure is commonly adopted. It was questioned whether the specific approach is sufficient for the studies targeting the bacterial domain, specifically nitrifiers, a group of autotrophic, slow growing microbes. In a controlled experiment mycorrhizal and non-mycorrhizal Plantago lanceolata were grown up in compartmentalized pots to study the AM effect on nitrification rates in the plant rhizosphere. Nitrification rates were assayed following an extensive 3-week bacterial equilibration step of the re-inoculated soil and a 13-week plant growth period in a controlled environment. Under these specific conditions, the nitrification potential levels at harvest were exceptionally low, and actual nitrification rates of the root compartment of non-mycorrhizal P. lanceolata were significantly lower than those of any other compartment. It is then argued that the specific effects should be attributed to the alleged higher growth rates of non-mycorrhizal plants that are known to occur early in the AM experiment. It is concluded that the specific experimental approach is not suitable for the study of microbes with slow growth rates.展开更多
基金Supported by 863 Program of China(2006AA03Z0453)NaturalScience Research Program of Higher Education of Jiangsu Province(09KJB230001)+1 种基金973 Program of China(2009CB724700)AndSchool Foundation of Jiangsu University(08JDG009)~~
文摘[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chromatography and fine chromatography. [Result]The optimized separation and purification conditions were that： Supernatant was treated by trichloroacetic acid, then by ultrafiltration desalt and anion exchange chromatography. Strong anion Q F. F. was better than weak anion DEAE F.F. The proper balanced solution was Tris-HCI （ pH 8.0）. The proper conductivity was 6 ms/cm. The maximum applied sample was 240 ATU/ml to matrix of strong anion Q F. F. This optimized procedure was magnified in strong anion exchange HiPrep 16/10Q with the 90% recovery and 70.2% purity. The purification of gel filtration of Sephacryl S-100 to hirudin was not relative to flow rate within certain scope. The application size of sample was 10 ml. The purity checked by HPLC was 95.1%, and the recovery was 93%, and the band of SDS-PAGE was single. [ Conclusion] The research provided the reference of the further industrialization separation and purification of hiruin.
基金Supported by National"973" Project(2011CB100400)~~
文摘[Objectives] This paper aims to explore the possibility to intercrop garlic with pomegranate tree to control pomegranate wilt.[Methods] Root exudates of garlic is cultivated in 1/5 concentration of MS solution and distilled water is examined in lab to test their effect to growth of mycelia of pomegranate wilt pathogen（Ceratocystis fimbriata）and multiplication of Bacillus subtilis.[Results] The result shows that garlic root exudates whatever cultivated in MS solution or distilled water could not inhibit or promote mycelia growth of C.fimbriata.However,garlic root exudates cultivated in both methods effectively promote multiplication of B.subtilis.[Conclusions] It is suggested that intercropping garlic with pomegranate tree by combining application B.subtilis could be a promising way to prevent pomegranate wilt spread in practice.
文摘Two plastid division genes, NtFtsZ1 and NtFtsZ2 isolated from Nicotiana tabacum L. were fused with gfp and expressed in Escherichia coli . The regular localizations of full length NtFtsZs∶GFP along the filamentous bacteria indicated that the NtFtsZs could recognize the potential division sites in E. coli and be polymerized with heterogeneous FtsZ from bacteria. The overexpression of NtFtsZs ∶ gfp inhibited the division of host strain cells and resulted in the long filamentous bacterial morphology. These results suggested that eukaryotic ftsZs have similar function to their prokaryotic homologs. Meanwhile, the different deletions of motifs of NtFtsZs are also employed to investigate the functions of these proteins in E. coli . The results showed that the C_terminal domains of NtFtsZs were related to the correct localization of NtFtsZs in E. coli and the N_terminal domains of NtFtsZs were responsible for the polymerization of homogeneous and heterogeneous FtsZ proteins. The significance of these results in understanding the functions of NtFtsZs in plastid division were discussed.
基金supported by the National Science and Technology Support Program (No. 2011BAD14B04)Project of Shandong Province Higher Educational Science and Technology Program (J14LE59)+1 种基金Applied & Basic Research Foundation of Qingdao (No. 12-1-4-3(3)-jch)Science & Technology Project of AQSIQ (No. 2012IK176)
文摘In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY- 3, which produced a bacteriocin-like substance （BLS）, was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16S rDNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria （including Staphylococcus aureus and B. subtillis） and some fimgi （including Penicillium glaucum, Aspergillus niger and Aspergillus flavus）. The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100~C for 60 min without losing any activity and stable over a wide pH range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid （HC1） and gel filteration （Sephadex G-100）. SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 kDa. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms.
基金Supported by a PhD fellowship from the Chloros trust
文摘The vast majority of herbaceous plants engage into arbuscular mycorrhizal (AM) symbioses and consideration of their mycorrhizal status should be embodied in studies of plant-microbe interactions. To establish reliable AM contrasts, however, a sterilized re-inoculation procedure is commonly adopted. It was questioned whether the specific approach is sufficient for the studies targeting the bacterial domain, specifically nitrifiers, a group of autotrophic, slow growing microbes. In a controlled experiment mycorrhizal and non-mycorrhizal Plantago lanceolata were grown up in compartmentalized pots to study the AM effect on nitrification rates in the plant rhizosphere. Nitrification rates were assayed following an extensive 3-week bacterial equilibration step of the re-inoculated soil and a 13-week plant growth period in a controlled environment. Under these specific conditions, the nitrification potential levels at harvest were exceptionally low, and actual nitrification rates of the root compartment of non-mycorrhizal P. lanceolata were significantly lower than those of any other compartment. It is then argued that the specific effects should be attributed to the alleged higher growth rates of non-mycorrhizal plants that are known to occur early in the AM experiment. It is concluded that the specific experimental approach is not suitable for the study of microbes with slow growth rates.
