The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeas...The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone a-factor （MFals）, and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction （PCR）-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae （SC-βG） was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A （PrA） activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/（h·ml） after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.展开更多
煤气化工艺作为煤制天然气的重要环节,是实现煤炭资源清洁利用的关键。基于化工流程模拟软件Aspen Plus建立BGL(British Gas Lurgi)炉煤气化工艺,以合成气有效成分、制气效率及水蒸气分解率为评价指标,考察常规气化技术中气化剂操作参...煤气化工艺作为煤制天然气的重要环节,是实现煤炭资源清洁利用的关键。基于化工流程模拟软件Aspen Plus建立BGL(British Gas Lurgi)炉煤气化工艺,以合成气有效成分、制气效率及水蒸气分解率为评价指标,考察常规气化技术中气化剂操作参数对工艺的耦合作用,得出最优气化剂组成及预热温度;同时,为解决煤气化过程中CO_2排放量大及氢碳比较低的问题,分别引入CO_2、CH_4对气化剂进行改进,进一步优化气化剂组成。结果表明:在常规气化技术中,m_(O_2)/m_(Coal)=0.33、m_(H_2O)/m_(Coal)=0.14为最优气化剂组成,220℃为最优气化剂预热温度;在改进气化技术中,m_(CO_2)/m_(H_2O)=0.18,m_(CH_4)/m_(H_2O)=0.08为最优气化剂组成。展开更多
基金the National Hi-Tech Research and Develop-ment Program (863) of China (No. 2007AA10Z315)the Natural Science Foundation of Zhejiang Province, China (No. Z304076)
文摘The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone a-factor （MFals）, and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction （PCR）-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae （SC-βG） was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A （PrA） activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/（h·ml） after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.
文摘煤气化工艺作为煤制天然气的重要环节,是实现煤炭资源清洁利用的关键。基于化工流程模拟软件Aspen Plus建立BGL(British Gas Lurgi)炉煤气化工艺,以合成气有效成分、制气效率及水蒸气分解率为评价指标,考察常规气化技术中气化剂操作参数对工艺的耦合作用,得出最优气化剂组成及预热温度;同时,为解决煤气化过程中CO_2排放量大及氢碳比较低的问题,分别引入CO_2、CH_4对气化剂进行改进,进一步优化气化剂组成。结果表明:在常规气化技术中,m_(O_2)/m_(Coal)=0.33、m_(H_2O)/m_(Coal)=0.14为最优气化剂组成,220℃为最优气化剂预热温度;在改进气化技术中,m_(CO_2)/m_(H_2O)=0.18,m_(CH_4)/m_(H_2O)=0.08为最优气化剂组成。
