Milk allergy is one of the most common food allergies,affecting 6%of young children,andβ-lactoglobulin(β-LG)is the main milk allergen.Clostridium tyrobutyricum Z816 was selected for the degradation ofβ-LG,which was...Milk allergy is one of the most common food allergies,affecting 6%of young children,andβ-lactoglobulin(β-LG)is the main milk allergen.Clostridium tyrobutyricum Z816 was selected for the degradation ofβ-LG,which was successfully reduced by about 90%using permeabilized bacteria under the optimized conditions.The hydrolyzed peptides were identified by liquid chromatography-tandem mass spectrometry(LC-MS/MS)and analyzed by molecular modeling,which indicated that C.tyrobutyricum Z816 could effectively degrade the antigenic epitopes ofβ-LG.Finally,the concentration and digestibility ofβ-LG in actual samples was quantified using enzyme-linked immunosorbent assay(ELISA)and gastrointestinal digestion simulation experiments.The results showed more than 92%ofβ-LG in actual samples was hydrolyzed,and the gastric and total digestibility of whey protein isolate(WPI)was improved by 85.96%and 64.51%,respectively.Therefore,C.tyrobutyricum Z816 offers an effective method to degradeβ-LG and reduce the occurrence of milk allergies,which has great significance for the development of hypoallergenic dairy products.展开更多
The objective of this study was to improve the production of butyric acid by process optimization using the metabolically engineered mutant of Clostridium tyrobutyricum (PAK-Em). First, the free-cell fermentation at...The objective of this study was to improve the production of butyric acid by process optimization using the metabolically engineered mutant of Clostridium tyrobutyricum (PAK-Em). First, the free-cell fermentation at pH 6.0 produced butyric acid with concentration of 38.44 g/L and yield of 0.42 g/g. Second, the immobilized- cell fermentations using fibrous-bed bioreactor (FBB) were run at pHs of 5.0, 5.5, 6.0, 6.5 and 7.0 to optimize fermentation process and improve the butyric acid production. It was found that the highest titer of butyric acid, 63.02g/L, was achieved at pH 6.5. Finally, the metabolic flux balance analysis was performed to inves- tigate the carbon rebalance in C. tyrobutyricum. The results show both gene manipulation and fermentation pH change redistribute carbon between biomass, acetic acid and butyric acid. This study demonstrated that high butyric acid production could be obtained by integrating metabolic engineering and fermentation process optimization.展开更多
为有效利用固态废弃毕赤酵母,高效发酵生产丁酸,建立了以80 g/L玉米淀粉为基础培养基、以废弃毕赤酵母处理液为高效有机氮源,连续补加葡萄糖的丁酸发酵工艺。7 L厌氧发酵罐下,发酵20h,先以1∶4的比例添加250~400 g/L规格的废弃酵母处理...为有效利用固态废弃毕赤酵母,高效发酵生产丁酸,建立了以80 g/L玉米淀粉为基础培养基、以废弃毕赤酵母处理液为高效有机氮源,连续补加葡萄糖的丁酸发酵工艺。7 L厌氧发酵罐下,发酵20h,先以1∶4的比例添加250~400 g/L规格的废弃酵母处理液替代昂贵的发酵用复合培养基,再连续补加葡萄糖浓缩液。最终的丁酸质量浓度为45 g/L、得率为40%,丁酸占总酸的比率(butyric acid ratio over total organic acids,B/TA)≥0.90的较高水平。该工艺可以有效地利用来自于废弃酵母处理液中的氨基酸,提高细胞生长速度和浓度,增强丁酸合成所依赖的NADH再生速度,间接改善了丁酸发酵性能。与此同时,实现了废弃生物质的有效利用和资源化。展开更多
利用马尔可夫链蒙特卡罗(Markov Chain Monte Carlo,MCMC)方法估计Logistic回归模型中的参数,就是要构造一个以参数的后验分布为其平稳分布的非周期不可约的马尔可夫链,然后用该平稳分布中抽出的样本点计算蒙特卡罗积分。上述理论方法...利用马尔可夫链蒙特卡罗(Markov Chain Monte Carlo,MCMC)方法估计Logistic回归模型中的参数,就是要构造一个以参数的后验分布为其平稳分布的非周期不可约的马尔可夫链,然后用该平稳分布中抽出的样本点计算蒙特卡罗积分。上述理论方法可以解决实验样本数据由于存在定和约束和多重共线性、在进行经典的logistic回归建模时的困难问题。基于此方法,研究了丁酸梭菌株对于给定辐照区间剂量的应答趋势,用模型挖掘数据所隐含的内在信息并导出了Logistic回归模型参数的贝叶斯框架下的50%,90%,95%和99%的置信区间。结果表明,运用Logistic与马尔可夫链耦合模型在有关给定辐射剂量对于微生物作用效果问题的logistic回归建模中具有较大的科学性与很好的使用性,从而可以为辐照诱变处理微生物制定辐照剂量区提供理论支持和回归技术借鉴。展开更多
基金supported by the National Key R&D Program of China(2017YFC1600404)the National Natural Science Foundation of China(31922070,22008114)the Natural Science Foundation of Jiangsu Province(BK20180038,BK20200684)。
文摘Milk allergy is one of the most common food allergies,affecting 6%of young children,andβ-lactoglobulin(β-LG)is the main milk allergen.Clostridium tyrobutyricum Z816 was selected for the degradation ofβ-LG,which was successfully reduced by about 90%using permeabilized bacteria under the optimized conditions.The hydrolyzed peptides were identified by liquid chromatography-tandem mass spectrometry(LC-MS/MS)and analyzed by molecular modeling,which indicated that C.tyrobutyricum Z816 could effectively degrade the antigenic epitopes ofβ-LG.Finally,the concentration and digestibility ofβ-LG in actual samples was quantified using enzyme-linked immunosorbent assay(ELISA)and gastrointestinal digestion simulation experiments.The results showed more than 92%ofβ-LG in actual samples was hydrolyzed,and the gastric and total digestibility of whey protein isolate(WPI)was improved by 85.96%and 64.51%,respectively.Therefore,C.tyrobutyricum Z816 offers an effective method to degradeβ-LG and reduce the occurrence of milk allergies,which has great significance for the development of hypoallergenic dairy products.
文摘The objective of this study was to improve the production of butyric acid by process optimization using the metabolically engineered mutant of Clostridium tyrobutyricum (PAK-Em). First, the free-cell fermentation at pH 6.0 produced butyric acid with concentration of 38.44 g/L and yield of 0.42 g/g. Second, the immobilized- cell fermentations using fibrous-bed bioreactor (FBB) were run at pHs of 5.0, 5.5, 6.0, 6.5 and 7.0 to optimize fermentation process and improve the butyric acid production. It was found that the highest titer of butyric acid, 63.02g/L, was achieved at pH 6.5. Finally, the metabolic flux balance analysis was performed to inves- tigate the carbon rebalance in C. tyrobutyricum. The results show both gene manipulation and fermentation pH change redistribute carbon between biomass, acetic acid and butyric acid. This study demonstrated that high butyric acid production could be obtained by integrating metabolic engineering and fermentation process optimization.
文摘为有效利用固态废弃毕赤酵母,高效发酵生产丁酸,建立了以80 g/L玉米淀粉为基础培养基、以废弃毕赤酵母处理液为高效有机氮源,连续补加葡萄糖的丁酸发酵工艺。7 L厌氧发酵罐下,发酵20h,先以1∶4的比例添加250~400 g/L规格的废弃酵母处理液替代昂贵的发酵用复合培养基,再连续补加葡萄糖浓缩液。最终的丁酸质量浓度为45 g/L、得率为40%,丁酸占总酸的比率(butyric acid ratio over total organic acids,B/TA)≥0.90的较高水平。该工艺可以有效地利用来自于废弃酵母处理液中的氨基酸,提高细胞生长速度和浓度,增强丁酸合成所依赖的NADH再生速度,间接改善了丁酸发酵性能。与此同时,实现了废弃生物质的有效利用和资源化。
文摘利用马尔可夫链蒙特卡罗(Markov Chain Monte Carlo,MCMC)方法估计Logistic回归模型中的参数,就是要构造一个以参数的后验分布为其平稳分布的非周期不可约的马尔可夫链,然后用该平稳分布中抽出的样本点计算蒙特卡罗积分。上述理论方法可以解决实验样本数据由于存在定和约束和多重共线性、在进行经典的logistic回归建模时的困难问题。基于此方法,研究了丁酸梭菌株对于给定辐照区间剂量的应答趋势,用模型挖掘数据所隐含的内在信息并导出了Logistic回归模型参数的贝叶斯框架下的50%,90%,95%和99%的置信区间。结果表明,运用Logistic与马尔可夫链耦合模型在有关给定辐射剂量对于微生物作用效果问题的logistic回归建模中具有较大的科学性与很好的使用性,从而可以为辐照诱变处理微生物制定辐照剂量区提供理论支持和回归技术借鉴。