建立了一种快速、简单、准确的牛奶蛋白改性纤维鉴定方法。使用ESI(Electron Spray Ionization)/TOF(Time of Flight)质谱对牛奶蛋白改性纤维胰蛋白酶解肽进行分析,确定了牛奶蛋白改性纤维特征肽段,使用该特征肽段制备得到特异性单克隆...建立了一种快速、简单、准确的牛奶蛋白改性纤维鉴定方法。使用ESI(Electron Spray Ionization)/TOF(Time of Flight)质谱对牛奶蛋白改性纤维胰蛋白酶解肽进行分析,确定了牛奶蛋白改性纤维特征肽段,使用该特征肽段制备得到特异性单克隆抗体;将酪蛋白包被于T线,建立了针对牛奶蛋白改性纤维的免疫层析快速鉴定方法。该方法以7M尿素为提取剂,检出限为氨基酸质量分数为1.00%的牛奶蛋白改性纤维(相当于混纺4.00%牛奶蛋白改性纤维的纱线),检测时间不超过20min(含前处理)。该方法无需使用分析仪器,对常见的纤维没有非特异性识别,纤维的染色不会对检测结果造成干扰。展开更多
This study almed to screen a highIy efficient ceI uIose-degrading compIex microbial system from soiI and rotten straw and then study its appIication to natural ceI uIose. The isoIated stralns were preIiminariIy screen...This study almed to screen a highIy efficient ceI uIose-degrading compIex microbial system from soiI and rotten straw and then study its appIication to natural ceI uIose. The isoIated stralns were preIiminariIy screened with Congo red stalning and the ceI uIases activities were determined with DNS method. The non-antagonis-tic highIy efficient ceI uIos-degrading stralns were seIected and cuItured combinedIy for deveIoping a ceI uIose-degrading compIex microbial system. The resuIts showed that the CMC enzyme activity of the mixed cuIture of the three fungi stralns was higher than those of the cuItures of the three singIe stralns. The morphoIogical and moIecuIar bioIogical identification indicated that F1 was Botryosphaeria, F2 was Rhi-zopus oryzae, and F5 was Fusarium oxysporum. When straw was used as carbon source, the CMC enzyme activities of F1, F2 and F5 were 39.2, 31.4 and 40.6 IU/mI, respectiveIy; whiIe the CMC enzyme activity of the mixed cuIture of F1＋F2＋F5 was 50.12 IU/mI, which was increased by 23% compared to that of the singIe straln of F5. The experimental resuIts indicated that the ceI uIose-degrading effect of the compIex microbial system was better than those of the singIe fungi stralns, and the singIe stralns of F1, F2 and F5 had certaln deveIopment potentials.展开更多
[Objective] The research aimed to lay the foundation for high-efficiency biological degradation microbial inoculums. [Method] under the 60℃ temperature the cellulolytic microbes in pig manure were isolated and determ...[Objective] The research aimed to lay the foundation for high-efficiency biological degradation microbial inoculums. [Method] under the 60℃ temperature the cellulolytic microbes in pig manure were isolated and determined the CMCase and Fpase,then proceeded the 16S rRNA gene analysis. [Result]The results showed that:BC1 and BC3 strains characterized higher carboxymethyl cellulose enzyme activity and higher filter paper activity,but their difference was small,then the 16S rDNA sequence of BC1 and BC3 strains were related to pseudomonas sp. (98% and 99% similarities,respectively).[Conclusion] the experiment laid foundation for high-efficiency biological heating agent.展开更多
In this paper, in order to get the target microbe which has high cellulose bio-degradation ability, we collected soil samples from environment rich in cellulose. Carboxymethyl cellulose sodium gel plate and filter pap...In this paper, in order to get the target microbe which has high cellulose bio-degradation ability, we collected soil samples from environment rich in cellulose. Carboxymethyl cellulose sodium gel plate and filter paper plate were used for microbe screening, and finally we got a strain named MI which has the highest cellulase producing ability, and it has been identified as Penicilium oxalicum by morphological and molecular biological identification. We have studied cellulose degradation ability of this strain under different conditions such as pH value, temperature and inoculating time. The results showed that CMCase which belongs to acid enzyme, could achieve its maximum enzyme activity 489.96 U/ml when the optimal pH value was 4. The FPAase could achieve its maximum enzyme activity 1,595.45 U/ml when the optimal pH value was 4, too. The optimal ferment temperature of CMCase is 50℃ when achieved the maximum of enzyme activity at the second day, whereas the FPAase could reach its tip top on the third day, So the optimal ferment temperature of FPAase is also 50℃.展开更多
The discovery of new, highly active, biomass-degrading enzymes is important to the development of a sustainable biofuels industry. Dictyoglomus turgidum, a thermophilic, anaerobic eubacterium that ferments cellulose a...The discovery of new, highly active, biomass-degrading enzymes is important to the development of a sustainable biofuels industry. Dictyoglomus turgidum, a thermophilic, anaerobic eubacterium that ferments cellulose and produces ethanol and hydrogen, was chosen as a candidate to screen for novel enzymes. A novel thermostable endoglucanase, CelA, was identified and purified during screening of a shotgun library of Dic（yoglomus turgidum and subsequently subcloned and expressed in E. coli. The celA gene coding for a 312 amino acid protein showed low homology to proteins outside the genus Dictoglomi and lacked an apparent signal peptide. CelA had a broad substrate range, possessing both endo and exo activity on soluble and insoluble β-（1,4）-Iinked glucose-containing substrates as well as endo activity on soluble and insoluble β-（1,4）-linked mannose containing substrates. The specific activity of CelA was 226 U/rag using β-glucan, 66 U/mg using glucomannan, and 63 U/mg using CMC as substrates. The high temperature optimum of 70 ℃ to 80 ℃ and wide substrate range of the enzyme might make it an excellent tool for biomass degradation at high temperature.展开更多
文摘建立了一种快速、简单、准确的牛奶蛋白改性纤维鉴定方法。使用ESI(Electron Spray Ionization)/TOF(Time of Flight)质谱对牛奶蛋白改性纤维胰蛋白酶解肽进行分析,确定了牛奶蛋白改性纤维特征肽段,使用该特征肽段制备得到特异性单克隆抗体;将酪蛋白包被于T线,建立了针对牛奶蛋白改性纤维的免疫层析快速鉴定方法。该方法以7M尿素为提取剂,检出限为氨基酸质量分数为1.00%的牛奶蛋白改性纤维(相当于混纺4.00%牛奶蛋白改性纤维的纱线),检测时间不超过20min(含前处理)。该方法无需使用分析仪器,对常见的纤维没有非特异性识别,纤维的染色不会对检测结果造成干扰。
基金Supported by Key Program for International S&T Cooperation Projects of China(2012DFA30600)National Key Technology Research and Development Program(2012BAD29B06)Special Scientific Research Fund of Marine Public Welfare Profession of China(201305013)~~
文摘This study almed to screen a highIy efficient ceI uIose-degrading compIex microbial system from soiI and rotten straw and then study its appIication to natural ceI uIose. The isoIated stralns were preIiminariIy screened with Congo red stalning and the ceI uIases activities were determined with DNS method. The non-antagonis-tic highIy efficient ceI uIos-degrading stralns were seIected and cuItured combinedIy for deveIoping a ceI uIose-degrading compIex microbial system. The resuIts showed that the CMC enzyme activity of the mixed cuIture of the three fungi stralns was higher than those of the cuItures of the three singIe stralns. The morphoIogical and moIecuIar bioIogical identification indicated that F1 was Botryosphaeria, F2 was Rhi-zopus oryzae, and F5 was Fusarium oxysporum. When straw was used as carbon source, the CMC enzyme activities of F1, F2 and F5 were 39.2, 31.4 and 40.6 IU/mI, respectiveIy; whiIe the CMC enzyme activity of the mixed cuIture of F1＋F2＋F5 was 50.12 IU/mI, which was increased by 23% compared to that of the singIe straln of F5. The experimental resuIts indicated that the ceI uIose-degrading effect of the compIex microbial system was better than those of the singIe fungi stralns, and the singIe stralns of F1, F2 and F5 had certaln deveIopment potentials.
基金Supported by Foundation of Henan Educational Committee(2010B530001)~~
文摘[Objective] The research aimed to lay the foundation for high-efficiency biological degradation microbial inoculums. [Method] under the 60℃ temperature the cellulolytic microbes in pig manure were isolated and determined the CMCase and Fpase,then proceeded the 16S rRNA gene analysis. [Result]The results showed that:BC1 and BC3 strains characterized higher carboxymethyl cellulose enzyme activity and higher filter paper activity,but their difference was small,then the 16S rDNA sequence of BC1 and BC3 strains were related to pseudomonas sp. (98% and 99% similarities,respectively).[Conclusion] the experiment laid foundation for high-efficiency biological heating agent.
文摘In this paper, in order to get the target microbe which has high cellulose bio-degradation ability, we collected soil samples from environment rich in cellulose. Carboxymethyl cellulose sodium gel plate and filter paper plate were used for microbe screening, and finally we got a strain named MI which has the highest cellulase producing ability, and it has been identified as Penicilium oxalicum by morphological and molecular biological identification. We have studied cellulose degradation ability of this strain under different conditions such as pH value, temperature and inoculating time. The results showed that CMCase which belongs to acid enzyme, could achieve its maximum enzyme activity 489.96 U/ml when the optimal pH value was 4. The FPAase could achieve its maximum enzyme activity 1,595.45 U/ml when the optimal pH value was 4, too. The optimal ferment temperature of CMCase is 50℃ when achieved the maximum of enzyme activity at the second day, whereas the FPAase could reach its tip top on the third day, So the optimal ferment temperature of FPAase is also 50℃.
文摘The discovery of new, highly active, biomass-degrading enzymes is important to the development of a sustainable biofuels industry. Dictyoglomus turgidum, a thermophilic, anaerobic eubacterium that ferments cellulose and produces ethanol and hydrogen, was chosen as a candidate to screen for novel enzymes. A novel thermostable endoglucanase, CelA, was identified and purified during screening of a shotgun library of Dic（yoglomus turgidum and subsequently subcloned and expressed in E. coli. The celA gene coding for a 312 amino acid protein showed low homology to proteins outside the genus Dictoglomi and lacked an apparent signal peptide. CelA had a broad substrate range, possessing both endo and exo activity on soluble and insoluble β-（1,4）-Iinked glucose-containing substrates as well as endo activity on soluble and insoluble β-（1,4）-linked mannose containing substrates. The specific activity of CelA was 226 U/rag using β-glucan, 66 U/mg using glucomannan, and 63 U/mg using CMC as substrates. The high temperature optimum of 70 ℃ to 80 ℃ and wide substrate range of the enzyme might make it an excellent tool for biomass degradation at high temperature.
