Three angiotensin I converting enzyme(ACE) inhibition peptides were isolated from sandworm Sipunculus nudus protein hydrolysate prepared using protamex. Consecutive purification methods, including size exclusion chrom...Three angiotensin I converting enzyme(ACE) inhibition peptides were isolated from sandworm Sipunculus nudus protein hydrolysate prepared using protamex. Consecutive purification methods, including size exclusion chromatography and reverse-phase high performance liquid chromatography(RP-HPLC), were used to isolate the ACE inhibition peptides. The amino acid sequences of the peptides were identified as Ile-Asn-Asp, Val-Glu-Pro-Gly and Leu-Ala-Asp-Glu-Phe. The IC_(50) values of the purified peptides for ACE inhibition activity were 34.72 μmol L^(-1), 20.55 μmol L^(-1) and 22.77 μmol L^(-1), respectively. These results suggested that S. nudus proteins contain specific peptides that can be released by enzymatic hydrolysis. This study may provide an experimental basis for further systematic research, rational development and clinical utilization of sandworm resources.展开更多
Hydantoinase and N-carbamoylase play important rol es in the production of optically pure amino acids from racemic 5-monosubstitut ed hydantoins. In this report, hydantoinase and the N-carbamoylase from Burkh olderia ...Hydantoinase and N-carbamoylase play important rol es in the production of optically pure amino acids from racemic 5-monosubstitut ed hydantoins. In this report, hydantoinase and the N-carbamoylase from Burkh olderia cepecia.njut01 were purified to homogeneity by chromatography (Pharma cia Explorer 100 system). The substrate specificity, enantioselectivity, pH depe ndence of activity and temperature stability of the activity were characterized. The results show that the hydantoinase and N-carbamoylase induced from Burkh olderia cepecia.njut01 are both strict D-stereo selective enzymes. They both hydrolyze substrates with side chains containing aliphatic and aromatic res idues with higher activity and affinity toward aromatic than aliphatic substitu ted substrates. The hydantoinase is a homotetramer with subunit molecular weight near 52,000 and is active between pH 6.5 and 10 with an optimum near pH 9.0. The en zyme is active at temperatures up to 60°C, however,it appears instable at h ig her temperatures. The subunit molecular weight of N-carbamoylase is about 35KD. The N-carbamoylase is active in the pH range from 6.0 to 9.5. The optim-pH is 7.2 and the optimizing bioconversion temperature of the N-carbamyolase is 52 °C.展开更多
A number of higher plants are able to hyperaccumulate cadmium(Cd). However, it is unknown whether cadmium(Cd) plays a biological functional role in the carbonic anhydrase(CA) of hyperaccumulators. A hydroponic experim...A number of higher plants are able to hyperaccumulate cadmium(Cd). However, it is unknown whether cadmium(Cd) plays a biological functional role in the carbonic anhydrase(CA) of hyperaccumulators. A hydroponic experiment was conducted to explore the potentially physiological function of Cd in CA and the accumulation and tolerance of Cd in the Zn/Cd hyperaccumulator Picris divaricata Vant. P. divaricata was exposed to nutrient solutions with six Cd concentrations(0, 5, 10, 25, 50 and 75 μmol L^(-1)). After 12 d, plants were harvested for the analysis of plant biomass, Cd concentration and CA activity. The Cd concentrations in plant increased with the increasing Cd in nutrient solution, reaching 640 and 3 100 mg kg^(-1) in shoot and root, respectively, at the 75 μmol L^(-1) Cd treatment. Meanwhile, plant growth was enhanced by the Cd treatments at 5–25 μmol L^(-1), but it was significantly inhibited when the plants were exposed to solutions with higher Cd concerntrations(50 and 75 μmol L^(-1)). Exposure to Cd significantly increased the CA activity in P. divaricata, which reached a maximum value of 21.27 U mg^(-1) proteins at the 25 μmol L^(-1)Cd treatment, and the CA activity and shoot Cd concentration were positively correlated at solutions Cd of ≤ 25 μmol L^(-1). Moreover, two protein bands appeared on the denatured gel electrophoresis of purified CA, indicating that P. divaricata may have CA isomers with their respective molecular weights at around 60 and 55 k Da, at least one of which is Cd-bound. In addition, trace amounts of Cd in purified CA significantly increased with the supplied Cd concentration in nutrient solution(5–25 μmol L^(-1)). The results suggested that Cd may play a biological role by enhancing the activities and forming the active Cd-specific CA in the hyperaccumulator P. divaricata.展开更多
In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry...In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry experiments,16S rRNA and gyrB gene analysis,the strains LZ-10 and LZ-11 were identified as Bacillus subtilis.Adopted the method of ammonium sulfate,DEAE-52 anion purify enzyme,finally used polyacrylamide gel electrophoresis(SDS-PAGE)to detect molecular weight.The strain LZ-10 had an amylase activity of 123.3 U·mL^(-1),a purification factor of 6.8,a recovery rate of 69.5% and an optimal temperature of 50℃.The amylase activity of the strain LZ-11 was 59.91 U·mL^(-1),the purification factor was 4.5,the recovery rate was 60.5%,and the optimum temperature was 55℃.The commodity enzyme derived from Bacillus subtilis was 37.5 U·mL^(-1).The relative molecular weight of amylase activity from the two strains was 55 ku.Both thermal stability and pH stability were higher than those of commercialized amylase.展开更多
The introduction of functionalized magnetizable particles for the purification of enzymes or for the multi-use of pre-immobilized biocatalysts offers a great potential for time and cost savings in biotechnological pro...The introduction of functionalized magnetizable particles for the purification of enzymes or for the multi-use of pre-immobilized biocatalysts offers a great potential for time and cost savings in biotechnological process design. The selective separation of the magnetizable particles is performed for example by a high-gradient magnetic separator. In this study FEM and CFD simulations of the magnetic field and the fluid flow field within a filter chamber of a magnetic separator were carried out, to find an optimal separator design. The motion of virtual magnetizable particles was calculated with a one-way coupled Lagrangian approach in order to test many geometric and parametric variations in reduced time. It was found that a flow homogenisator smoothed the fluid flow, so that the linear velocity became nearly equal over the cross section in the direction of flow. Furthermore the retention of magnetizable particles increases with a high total edge length within the filter matrix.展开更多
基金supported by research grant of Guangxi Key Laboratory Traditional Chinese Medicine Quality Standards (No. GXGZZK201501)the Open Research Fund Program of Guangxi Key Laboratory of Marine Biotechnology (No. GLMBT-201407)+1 种基金partly supported by Shanghai Fengxian District Science and Technology Project (Nos. 20141001 and 20151205)Shanghai No. 6 People’s Medical Group Project and research project of Shanghai municipal health and Family Planning Commission (No. 201540027)
文摘Three angiotensin I converting enzyme(ACE) inhibition peptides were isolated from sandworm Sipunculus nudus protein hydrolysate prepared using protamex. Consecutive purification methods, including size exclusion chromatography and reverse-phase high performance liquid chromatography(RP-HPLC), were used to isolate the ACE inhibition peptides. The amino acid sequences of the peptides were identified as Ile-Asn-Asp, Val-Glu-Pro-Gly and Leu-Ala-Asp-Glu-Phe. The IC_(50) values of the purified peptides for ACE inhibition activity were 34.72 μmol L^(-1), 20.55 μmol L^(-1) and 22.77 μmol L^(-1), respectively. These results suggested that S. nudus proteins contain specific peptides that can be released by enzymatic hydrolysis. This study may provide an experimental basis for further systematic research, rational development and clinical utilization of sandworm resources.
文摘Hydantoinase and N-carbamoylase play important rol es in the production of optically pure amino acids from racemic 5-monosubstitut ed hydantoins. In this report, hydantoinase and the N-carbamoylase from Burkh olderia cepecia.njut01 were purified to homogeneity by chromatography (Pharma cia Explorer 100 system). The substrate specificity, enantioselectivity, pH depe ndence of activity and temperature stability of the activity were characterized. The results show that the hydantoinase and N-carbamoylase induced from Burkh olderia cepecia.njut01 are both strict D-stereo selective enzymes. They both hydrolyze substrates with side chains containing aliphatic and aromatic res idues with higher activity and affinity toward aromatic than aliphatic substitu ted substrates. The hydantoinase is a homotetramer with subunit molecular weight near 52,000 and is active between pH 6.5 and 10 with an optimum near pH 9.0. The en zyme is active at temperatures up to 60°C, however,it appears instable at h ig her temperatures. The subunit molecular weight of N-carbamoylase is about 35KD. The N-carbamoylase is active in the pH range from 6.0 to 9.5. The optim-pH is 7.2 and the optimizing bioconversion temperature of the N-carbamyolase is 52 °C.
基金supported by the Environmental Pollution Control and Remediation Technology Foundation of Guangdong Provincial Key Lab,China(No.2011K0002)
文摘A number of higher plants are able to hyperaccumulate cadmium(Cd). However, it is unknown whether cadmium(Cd) plays a biological functional role in the carbonic anhydrase(CA) of hyperaccumulators. A hydroponic experiment was conducted to explore the potentially physiological function of Cd in CA and the accumulation and tolerance of Cd in the Zn/Cd hyperaccumulator Picris divaricata Vant. P. divaricata was exposed to nutrient solutions with six Cd concentrations(0, 5, 10, 25, 50 and 75 μmol L^(-1)). After 12 d, plants were harvested for the analysis of plant biomass, Cd concentration and CA activity. The Cd concentrations in plant increased with the increasing Cd in nutrient solution, reaching 640 and 3 100 mg kg^(-1) in shoot and root, respectively, at the 75 μmol L^(-1) Cd treatment. Meanwhile, plant growth was enhanced by the Cd treatments at 5–25 μmol L^(-1), but it was significantly inhibited when the plants were exposed to solutions with higher Cd concerntrations(50 and 75 μmol L^(-1)). Exposure to Cd significantly increased the CA activity in P. divaricata, which reached a maximum value of 21.27 U mg^(-1) proteins at the 25 μmol L^(-1)Cd treatment, and the CA activity and shoot Cd concentration were positively correlated at solutions Cd of ≤ 25 μmol L^(-1). Moreover, two protein bands appeared on the denatured gel electrophoresis of purified CA, indicating that P. divaricata may have CA isomers with their respective molecular weights at around 60 and 55 k Da, at least one of which is Cd-bound. In addition, trace amounts of Cd in purified CA significantly increased with the supplied Cd concentration in nutrient solution(5–25 μmol L^(-1)). The results suggested that Cd may play a biological role by enhancing the activities and forming the active Cd-specific CA in the hyperaccumulator P. divaricata.
基金Supported by Young Doctor Fund of Gansu Education Department,China(2021QB-034)。
文摘In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry experiments,16S rRNA and gyrB gene analysis,the strains LZ-10 and LZ-11 were identified as Bacillus subtilis.Adopted the method of ammonium sulfate,DEAE-52 anion purify enzyme,finally used polyacrylamide gel electrophoresis(SDS-PAGE)to detect molecular weight.The strain LZ-10 had an amylase activity of 123.3 U·mL^(-1),a purification factor of 6.8,a recovery rate of 69.5% and an optimal temperature of 50℃.The amylase activity of the strain LZ-11 was 59.91 U·mL^(-1),the purification factor was 4.5,the recovery rate was 60.5%,and the optimum temperature was 55℃.The commodity enzyme derived from Bacillus subtilis was 37.5 U·mL^(-1).The relative molecular weight of amylase activity from the two strains was 55 ku.Both thermal stability and pH stability were higher than those of commercialized amylase.
文摘The introduction of functionalized magnetizable particles for the purification of enzymes or for the multi-use of pre-immobilized biocatalysts offers a great potential for time and cost savings in biotechnological process design. The selective separation of the magnetizable particles is performed for example by a high-gradient magnetic separator. In this study FEM and CFD simulations of the magnetic field and the fluid flow field within a filter chamber of a magnetic separator were carried out, to find an optimal separator design. The motion of virtual magnetizable particles was calculated with a one-way coupled Lagrangian approach in order to test many geometric and parametric variations in reduced time. It was found that a flow homogenisator smoothed the fluid flow, so that the linear velocity became nearly equal over the cross section in the direction of flow. Furthermore the retention of magnetizable particles increases with a high total edge length within the filter matrix.
