A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calci...A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calcined at 500 ℃, denoted as CLDH, was characterized by XRD, IR and BET surface area measurements. CLDH has a poor crystalline MgO-like structure with a high surface area and porosity. CLDH was used as a support for the immobilization of penicillin G acylase(PGA). The effect of varying the immobilization conditions, such as pH, contact time and the ratio of enzyme to support, on the activity of the immobilized enzyme in the hydrolysis of penicillin G has been studied. It was found that the activity of the immobilized enzyme decreased slightly with decreasing pH and reached a maximum after a contact time of 24 h. The activity of the immobilized enzyme increased with increasing the ratio of enzyme to support. It was found that the adsorption of PGA inhibited the expected reaction of CLDH with an aqueous medium to regenerate a LDH phase. Its original activity(36%) after 15 cycles of reuse of the immobilized enzyme was retained, but no further loss in the activity was observed.展开更多
为探究双酶不同酶解路线制得大米蛋白肽的性质差异,研究采用胰蛋白酶(A)和碱性蛋白酶(B)按不同酶解路线对大米蛋白进行酶解,制备了5种大米蛋白肽(A^(1)B^(1)、A^(1)B^(2)、A^(2)B^(1)、A^(1*)B^(2)、A^(2)B^(1*)),对其水解度、基本成分...为探究双酶不同酶解路线制得大米蛋白肽的性质差异,研究采用胰蛋白酶(A)和碱性蛋白酶(B)按不同酶解路线对大米蛋白进行酶解,制备了5种大米蛋白肽(A^(1)B^(1)、A^(1)B^(2)、A^(2)B^(1)、A^(1*)B^(2)、A^(2)B^(1*)),对其水解度、基本成分、氨基酸组成、微观结构、二级结构、分子量分布、风味和体外抗氧化活性等进行分析。结果表明,A^(2)B^(1)组的蛋白含量最高达到90.69%,肽含量高达72.73%;各路线的水解度大于17.60%,水解较为完全;微观结构均由不规则块状变为球体状,A1B2组和A^(1*)B^(2)组球体壁较厚,A^(2)B^(1)组和A^(2)B^(1*)组球体壁较薄,A^(1)B^(1)组为球体碎片;各路线的必需氨基酸含量比大米蛋白低,A1B2组必需氨基酸含量最高;二级结构以多种构象并存,各路线二级结构均以β-转角为主,占二级结构的44.62%~47.18%;各路线酶解产物多为低分子量的多肽,分子量低于5 k Da的多肽占92.09%~93.71%;A1B2样品鲜味最强,涩味最弱,A^(2)B^(1)样品咸味和苦味最弱;相比于A^(1)B^(1)组、A1B2组和A^(1*)B^(2)组,A^(2)B^(1)组和A^(2)B^(1*)组的抗氧化活性更强。通过对双酶不同酶解路线大米蛋白肽的性质研究,综合基本成分、肽含量、氨基酸组成、风味和抗氧化活性评价,A^(2)B^(1)组的品质最佳。展开更多
目的建立皮尔逊相关系数(Pearson correlation coefficient,PCC)和长短期记忆(long short term memory,LSTM)神经网络的反应液葡萄糖含量预测模型用以实时预测葡萄糖酸锌生产过程中反应液葡萄糖含量。方法通过葡萄糖酸锌制备实验,结合PC...目的建立皮尔逊相关系数(Pearson correlation coefficient,PCC)和长短期记忆(long short term memory,LSTM)神经网络的反应液葡萄糖含量预测模型用以实时预测葡萄糖酸锌生产过程中反应液葡萄糖含量。方法通过葡萄糖酸锌制备实验,结合PCC理论确定对反应液葡萄糖含量有较大影响的因素,对这些因素进行数据采集并将其作为神经网络的输入变量,采集反应液葡萄糖含量数据并进行处理,将其作为神经网络的输出变量,进而建立反向传播神经网络(backpropagation neural network,BP)和LSTM神经网络的反应液葡萄糖含量预测模型。结果通过100次模型迭代训练,对照BP反应液葡萄糖含量预测模型可以看出LSTM反应液葡萄糖含量预测模型在测试集的误差约为0.45%,误差较小,准确度较高。结论基于LSTM反应液葡萄糖含量预测模型显著提高了预测精度,相比现有检测方法更加智能高效,能够有效辅助生产进行。展开更多
Zhongshuang9, a new semi-winter Brassica napus variety with high resistance to Sclerotinia sclerotiorum and lodging, high-yield, double-low quality and extensive adaptability, was bred by multiple crossing and microsp...Zhongshuang9, a new semi-winter Brassica napus variety with high resistance to Sclerotinia sclerotiorum and lodging, high-yield, double-low quality and extensive adaptability, was bred by multiple crossing and microspore culture technique. It was registered and released in China in 2002. In regional trial of Hubei Province in China, Zhongshuang9 yielded 2 482. 2 kg ha-1 averagely in 2000 - 2002, 15. 33% higher than the control variety Zhongyou821. Erucic acid, glucosinolates and oil contents of Zhongshuang9 were 0.23%, 22.69 μmol g-1(in meal)and 42%, respectively. In field assessment of resistance to S. Sclerotiorum , the disease incidence and disease index of Zhongshuang9 averaged 13.31 % and 6.47, respectively, which were lower than those of Zhongyou821 by 28% and 36%, respectively. After inoculation of detached leaves with mycelia, the lesion size of Zhongshuang9 was 4. 709 cm2, which was significantly smaller than that of the mid-resistant variety Zhongyou821(5. 933 cm2). The stem lesion length of Zhongshuang9 after match-stick inoculation was 1.275 cm, which was significantly lower than that of Zhongyou821(1.943 cm). The possible mechanism of resistance to S. sclerotiorum was studied through comparing the activities of phenylalanine ammonia lyase(PAL), exo-chitinase, β-1, 3-glucanase, peroxidase(POD)and polyphenoloxidase(PPO)in Zhongshuang9 with those in other resistant, mid-resistant and susceptible cultivars.展开更多
目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌E...目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌EscherichiacoliBL21(DE3)中进行重组和异丙基-β-D-硫代半乳糖苷(isopropylβ-D-thiogalactoside,IPTG)诱导表达,经镍亲和层析纯化并鉴定AKR18A2的酶学性质,构建德沃斯氏菌(Devosiasp.)来源的DON降解酶QDDH和AKR18A2双酶作用体系,实现DON的高效降解。结果来自鞘氨醇单胞菌属(Sphingomonas sp.)的呕吐毒素降解酶(AKR18A2)由343个氨基酸组成。该酶属于醛酮还原酶超家族,45℃和pH7.0为最适反应条件。AKR18A2可在24h内降解15.42%DON,而双酶(QDDH和AKR18A2)协同作用4h后对DON的降解率可提高至98.02%。结论本研究鉴定了一种新型DON降解酶AKR18A2,并首次创新建立双酶联用体系进一步实现DON的高效降解。展开更多
In the present research,enzyme encapsulated hydrogels（single gels and double network gels）and enzyme immobilized magnetic beads,which allow high-throughput screening,were fabricated and evaluated in terms of the pre...In the present research,enzyme encapsulated hydrogels（single gels and double network gels）and enzyme immobilized magnetic beads,which allow high-throughput screening,were fabricated and evaluated in terms of the preservation,precision, and repeatability of enzyme activity.The fabricated gels and magnetic beads were analyzed in a 96-well microassay plate.Trypsin was successfully encapsulated in both types of gels and immobilized to the magnetic beads.However,pepsin,either encapsulated in the gels or immobilized to the magnetic beads,could not react with its substrates.The adaptability to various enzymes （e.g.,trypsin,β-glucuronidase,and CYP1A1）in the single gels and magnetic beads was superior to that in double network gels.However,the soak out of the enzymes was observed in the single gels.The double network gels could encapsulate trypsin,whereas the fabrication of the other enzymes（e.g.β-glucuronidase,CYP1A1,and pepsin）failed because of the inactivation of the enzymes by acryl amide and ammonium peroxodisulfate,which are the components of the gel formulation. The enzyme reaction in the magnetic beads exhibited the highest efficiency among the three fabrication methods.Furthermore, the stability of the enzymes immobilized to the magnetic beads was better than that fabricated by the other methods,and the activities of trypsin andβ-glucuronidase did not decline for up to one week.In addition,in the magnetic beads,the activities of trypsin andβ-glucuronidase can be well repeated.Hence,although the adaptability of the double network gels to various enzymes is currently limited,the efficiency of the enzyme encapsulation can be improved by optimizing the formulation of acryl amide gels.展开更多
基金Supported by the National Natural Science Foundation of China( No.2 99730 0 4 )
文摘A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calcined at 500 ℃, denoted as CLDH, was characterized by XRD, IR and BET surface area measurements. CLDH has a poor crystalline MgO-like structure with a high surface area and porosity. CLDH was used as a support for the immobilization of penicillin G acylase(PGA). The effect of varying the immobilization conditions, such as pH, contact time and the ratio of enzyme to support, on the activity of the immobilized enzyme in the hydrolysis of penicillin G has been studied. It was found that the activity of the immobilized enzyme decreased slightly with decreasing pH and reached a maximum after a contact time of 24 h. The activity of the immobilized enzyme increased with increasing the ratio of enzyme to support. It was found that the adsorption of PGA inhibited the expected reaction of CLDH with an aqueous medium to regenerate a LDH phase. Its original activity(36%) after 15 cycles of reuse of the immobilized enzyme was retained, but no further loss in the activity was observed.
文摘为探究双酶不同酶解路线制得大米蛋白肽的性质差异,研究采用胰蛋白酶(A)和碱性蛋白酶(B)按不同酶解路线对大米蛋白进行酶解,制备了5种大米蛋白肽(A^(1)B^(1)、A^(1)B^(2)、A^(2)B^(1)、A^(1*)B^(2)、A^(2)B^(1*)),对其水解度、基本成分、氨基酸组成、微观结构、二级结构、分子量分布、风味和体外抗氧化活性等进行分析。结果表明,A^(2)B^(1)组的蛋白含量最高达到90.69%,肽含量高达72.73%;各路线的水解度大于17.60%,水解较为完全;微观结构均由不规则块状变为球体状,A1B2组和A^(1*)B^(2)组球体壁较厚,A^(2)B^(1)组和A^(2)B^(1*)组球体壁较薄,A^(1)B^(1)组为球体碎片;各路线的必需氨基酸含量比大米蛋白低,A1B2组必需氨基酸含量最高;二级结构以多种构象并存,各路线二级结构均以β-转角为主,占二级结构的44.62%~47.18%;各路线酶解产物多为低分子量的多肽,分子量低于5 k Da的多肽占92.09%~93.71%;A1B2样品鲜味最强,涩味最弱,A^(2)B^(1)样品咸味和苦味最弱;相比于A^(1)B^(1)组、A1B2组和A^(1*)B^(2)组,A^(2)B^(1)组和A^(2)B^(1*)组的抗氧化活性更强。通过对双酶不同酶解路线大米蛋白肽的性质研究,综合基本成分、肽含量、氨基酸组成、风味和抗氧化活性评价,A^(2)B^(1)组的品质最佳。
文摘目的建立皮尔逊相关系数(Pearson correlation coefficient,PCC)和长短期记忆(long short term memory,LSTM)神经网络的反应液葡萄糖含量预测模型用以实时预测葡萄糖酸锌生产过程中反应液葡萄糖含量。方法通过葡萄糖酸锌制备实验,结合PCC理论确定对反应液葡萄糖含量有较大影响的因素,对这些因素进行数据采集并将其作为神经网络的输入变量,采集反应液葡萄糖含量数据并进行处理,将其作为神经网络的输出变量,进而建立反向传播神经网络(backpropagation neural network,BP)和LSTM神经网络的反应液葡萄糖含量预测模型。结果通过100次模型迭代训练,对照BP反应液葡萄糖含量预测模型可以看出LSTM反应液葡萄糖含量预测模型在测试集的误差约为0.45%,误差较小,准确度较高。结论基于LSTM反应液葡萄糖含量预测模型显著提高了预测精度,相比现有检测方法更加智能高效,能够有效辅助生产进行。
文摘Zhongshuang9, a new semi-winter Brassica napus variety with high resistance to Sclerotinia sclerotiorum and lodging, high-yield, double-low quality and extensive adaptability, was bred by multiple crossing and microspore culture technique. It was registered and released in China in 2002. In regional trial of Hubei Province in China, Zhongshuang9 yielded 2 482. 2 kg ha-1 averagely in 2000 - 2002, 15. 33% higher than the control variety Zhongyou821. Erucic acid, glucosinolates and oil contents of Zhongshuang9 were 0.23%, 22.69 μmol g-1(in meal)and 42%, respectively. In field assessment of resistance to S. Sclerotiorum , the disease incidence and disease index of Zhongshuang9 averaged 13.31 % and 6.47, respectively, which were lower than those of Zhongyou821 by 28% and 36%, respectively. After inoculation of detached leaves with mycelia, the lesion size of Zhongshuang9 was 4. 709 cm2, which was significantly smaller than that of the mid-resistant variety Zhongyou821(5. 933 cm2). The stem lesion length of Zhongshuang9 after match-stick inoculation was 1.275 cm, which was significantly lower than that of Zhongyou821(1.943 cm). The possible mechanism of resistance to S. sclerotiorum was studied through comparing the activities of phenylalanine ammonia lyase(PAL), exo-chitinase, β-1, 3-glucanase, peroxidase(POD)and polyphenoloxidase(PPO)in Zhongshuang9 with those in other resistant, mid-resistant and susceptible cultivars.
文摘目的挖掘呕吐毒素(deoxynivalenol,DON)降解酶,应用双酶体系实现呕吐毒素的高效降解。方法利用生物信息学技术从国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库筛选潜在DON降解酶AKR18A2,在大肠杆菌EscherichiacoliBL21(DE3)中进行重组和异丙基-β-D-硫代半乳糖苷(isopropylβ-D-thiogalactoside,IPTG)诱导表达,经镍亲和层析纯化并鉴定AKR18A2的酶学性质,构建德沃斯氏菌(Devosiasp.)来源的DON降解酶QDDH和AKR18A2双酶作用体系,实现DON的高效降解。结果来自鞘氨醇单胞菌属(Sphingomonas sp.)的呕吐毒素降解酶(AKR18A2)由343个氨基酸组成。该酶属于醛酮还原酶超家族,45℃和pH7.0为最适反应条件。AKR18A2可在24h内降解15.42%DON,而双酶(QDDH和AKR18A2)协同作用4h后对DON的降解率可提高至98.02%。结论本研究鉴定了一种新型DON降解酶AKR18A2,并首次创新建立双酶联用体系进一步实现DON的高效降解。
基金The Global COE Program from the Ministry of Education,Science,Sports,and Culture of Japan.
文摘In the present research,enzyme encapsulated hydrogels（single gels and double network gels）and enzyme immobilized magnetic beads,which allow high-throughput screening,were fabricated and evaluated in terms of the preservation,precision, and repeatability of enzyme activity.The fabricated gels and magnetic beads were analyzed in a 96-well microassay plate.Trypsin was successfully encapsulated in both types of gels and immobilized to the magnetic beads.However,pepsin,either encapsulated in the gels or immobilized to the magnetic beads,could not react with its substrates.The adaptability to various enzymes （e.g.,trypsin,β-glucuronidase,and CYP1A1）in the single gels and magnetic beads was superior to that in double network gels.However,the soak out of the enzymes was observed in the single gels.The double network gels could encapsulate trypsin,whereas the fabrication of the other enzymes（e.g.β-glucuronidase,CYP1A1,and pepsin）failed because of the inactivation of the enzymes by acryl amide and ammonium peroxodisulfate,which are the components of the gel formulation. The enzyme reaction in the magnetic beads exhibited the highest efficiency among the three fabrication methods.Furthermore, the stability of the enzymes immobilized to the magnetic beads was better than that fabricated by the other methods,and the activities of trypsin andβ-glucuronidase did not decline for up to one week.In addition,in the magnetic beads,the activities of trypsin andβ-glucuronidase can be well repeated.Hence,although the adaptability of the double network gels to various enzymes is currently limited,the efficiency of the enzyme encapsulation can be improved by optimizing the formulation of acryl amide gels.
