ABSTRACT
Abstract We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P ≤ 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.
Resumo Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P ≤ 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.
Subject(s)
Animals , Body Composition , FishesABSTRACT
A ninety days nutritional trial was directed to explore the effects of dietary chromium on body composition, gut enzyme activity and physiological status of Cirrhinus mrigala by using G & NG corn. Six experimental diets were prepared by using different levels of chromium chloride hexahydrate (0, 0.2, 0.4 mg/kg, each with G & NG corn). For this experimental trial, 480 fingerlings, irrespective of sex were distributed in six aquariums each with replicate. Results revealed that gelatinized corn along with increasing level of Cr2Cl3.6H2O have a positive impact upon body composition of fish. Hematology was positively correlated with chromium chloride hexahydrate supplementation in gelatinized corn. Amylase gut enzyme also showed significant (P<0.05) increase in group fed with chromium chloride hexahydrate supplemented diet (G corn). However, corn with chromium chloride hexahydrate supplementation did not revealed any significant impact on gut protease enzyme activity. From these results it can be concluded that both chromium chloride hexahydrate and gelatinized corn in fish feed are very beneficial to improve body composition, enzymes activity and physiological health status of fish.
Um ensaio nutricional de 90 dias foi dirigido para explorar os efeitos do cromo dietético na composição corporal, atividade enzimática intestinal e estado fisiológico de Cirrhinus mrigala usando milho G & NG. Seis dietas experimentais foram preparadas usando diferentes níveis de cloreto de cromo hexa-hidratado (0, 0,2, 0,4 mg/kg, cada um com milho G e NG). Para este ensaio experimental, 480 alevinos, independentemente do sexo, foram distribuídos em seis aquários, cada um com réplicas. Os resultados revelaram que o milho gelatinizado, juntamente com o aumento do nível de Cr2Cl3.6H2O, tem um impacto positivo na composição corporal dos peixes. A hematologia foi positivamente correlacionada com a suplementação de cloreto de cromo hexa-hidratado em milho gelatinizado. A enzima amilase intestinal também apresentou aumento significativo (P <0,05) no grupo alimentado com dieta suplementada com cloreto de cromo hexa-hidratado (milho G). No entanto, o milho com suplementação de hexahidrato de cloreto de cromo não revelou nenhum impacto significativo na atividade da enzima protease intestinal. Desses resultados, pode-se concluir que tanto o cloreto de cromo hexa-hidratado quanto o milho gelatinizado na alimentação de peixes são muito benéficos para melhorar a composição corporal, a atividade de enzimas e o estado fisiológico de saúde dos peixes.
Subject(s)
Animals , Dietary Carbohydrates/administration & dosage , Chromium/administration & dosage , Cyprinidae/growth & development , Cyprinidae/physiology , Cyprinidae/metabolism , Cyprinidae/blood , Zea maysABSTRACT
We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P ≤ 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.
Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P ≤ 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.
Subject(s)
Animals , Enzyme Activation , Cyprinidae/growth & development , Peptide Hydrolases/metabolismABSTRACT
Abstract A ninety days nutritional trial was directed to explore the effects of dietary chromium on body composition, gut enzyme activity and physiological status of Cirrhinus mrigala by using G & NG corn. Six experimental diets were prepared by using different levels of chromium chloride hexahydrate (0, 0.2, 0.4 mg/kg, each with G & NG corn). For this experimental trial, 480 fingerlings, irrespective of sex were distributed in six aquariums each with replicate. Results revealed that gelatinized corn along with increasing level of Cr2Cl3.6H2O have a positive impact upon body composition of fish. Hematology was positively correlated with chromium chloride hexahydrate supplementation in gelatinized corn. Amylase gut enzyme also showed significant (P 0.05) increase in group fed with chromium chloride hexahydrate supplemented diet (G corn). However, corn with chromium chloride hexahydrate supplementation did not revealed any significant impact on gut protease enzyme activity. From these results it can be concluded that both chromium chloride hexahydrate and gelatinized corn in fish feed are very beneficial to improve body composition, enzymes activity and physiological health status of fish.
Resumo Um ensaio nutricional de 90 dias foi dirigido para explorar os efeitos do cromo dietético na composição corporal, atividade enzimática intestinal e estado fisiológico de Cirrhinus mrigala usando milho G & NG. Seis dietas experimentais foram preparadas usando diferentes níveis de cloreto de cromo hexa-hidratado (0, 0,2, 0,4 mg/kg, cada um com milho G e NG). Para este ensaio experimental, 480 alevinos, independentemente do sexo, foram distribuídos em seis aquários, cada um com réplicas. Os resultados revelaram que o milho gelatinizado, juntamente com o aumento do nível de Cr2Cl3.6H2O, tem um impacto positivo na composição corporal dos peixes. A hematologia foi positivamente correlacionada com a suplementação de cloreto de cromo hexa-hidratado em milho gelatinizado. A enzima amilase intestinal também apresentou aumento significativo (P 0,05) no grupo alimentado com dieta suplementada com cloreto de cromo hexa-hidratado (milho G). No entanto, o milho com suplementação de hexahidrato de cloreto de cromo não revelou nenhum impacto significativo na atividade da enzima protease intestinal. Desses resultados, pode-se concluir que tanto o cloreto de cromo hexa-hidratado quanto o milho gelatinizado na alimentação de peixes são muito benéficos para melhorar a composição corporal, a atividade de enzimas e o estado fisiológico de saúde dos peixes.
ABSTRACT
Abstract We have evaluated the effects of different fish feeds on the body composition, growth, and enzyme activities of Labeo rohita (Rohu). In total, 240 fishes between the average weights of 24.77±2.15g were studied. The treatments were applied in a completely randomized design, with 4 treatments of 60 fishes each. Treatments consisted of four different fish feeds [Oryza (T1), AMG (T2), Aqua (T3), and Supreme (T4)]. Body composition, growth performance, and enzyme activities were evaluated. There was a significant variation in performance of fishes fed with different type of feed; as fishes having Oryza feed showed the highest weight gain, specific growth rate (SGR), and best feed conversion ratio (FCR) as compared to other groups that were considered to be significant (P 0.05). High net weight gain was obtained in T4 when compared with T2 and T3. FCR value of T4 was less than T1 but higher than T2, T3 and T2, which showed the lowest values. The specific growth rate was recorded as average in T4, but T2 led a high SGR than T3. Similarly, crude protein level and digestive enzymes activity was recorded significantly highest in fed with Oryza (T1) as compared to AMG (T2), Aqua (T3), and Supreme (T4). Water quality parameters were recorded significant in all treatments except pH and DO of treatment (T1), significantly different from other treatments. It was concluded that Rohu (Labeo rohita) could show a promising growth rate and protease enzyme activity when fed with the Oryza feed of 25% protein.
Resumo Avaliamos os efeitos de diferentes alimentos para peixes em relação à composição corporal, crescimento e atividades enzimáticas de Labeo rohita (Rohu). No total, foram estudados 240 peixes com pesos médios de 24,77 ± 2,15 g. Os tratamentos foram aplicados em delineamento inteiramente casualizado, com quatro tratamentos de 60 peixes cada. Os tratamentos consistiram em quatro alimentos diferentes para peixes: Oryza (T1), AMG (T2), Aqua (T3) e Supreme (T4). Foram avaliados a composição corporal, o desempenho de crescimento e as atividades enzimáticas. Houve uma variação significativa no desempenho dos peixes alimentados com diferentes tipos de ração. Peixes com alimentação Oryza apresentaram maior ganho de peso, taxa de crescimento específico (SGR) e melhor taxa de conversão alimentar (FCR) em comparação com outros grupos que foram considerados significativos (P 0,05). Elevado ganho de peso líquido foi obtido em T4 quando comparado com T2 e T3. O valor da FCR de T4 foi menor que T1, mas maior que T2 e T3, que apresentaram os menores valores. A taxa de crescimento específico foi registrada como média em T4, mas T2 teve uma SGR alta do que T3. Da mesma forma, o nível de proteína bruta e a atividade das enzimas digestivas foram registrados significativamente mais altos nos peixes alimentados com Oryza (T1) em comparação com AMG (T2), Aqua (T3) e Supreme (T4). Os parâmetros de qualidade da água foram registrados como significativos em todos os tratamentos, exceto pH e OD do tratamento (T1), significativamente diferente dos demais tratamentos. Concluiu-se que Rohu (Labeo rohita) pode apresentar uma taxa de crescimento promissora e atividade da enzima protease quando alimentado com Oryza de 25% de proteína.
ABSTRACT
Dihydroflavonol 4-reductase (DFR) plays an essential role in the biosynthesis of anthocyanin and regulation of plant flower color. Based on the transcriptome data of Cistanche tubulosa (Schenk) Wight, a full-length cDNA sequence of CtDFR gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR). CtDFR contains an open reading frame (ORF) of 1 263 bp which encodes 420 amino acids with a predicted molecular weight of 47.5 kDa. The sequence analysis showed that CtDFR contains a nicotinamide adenine dinucleotide phosphate (NADPH) binding domain and a specific substrate binding domain. The expression analysis indicated that CtDFR was highly expressed in red and purple flowers, and the relative expression levels were 4.04 and 19.37 times higher than those of white flowers, respectively. The recombinant CtDFR protein was expressed in E.coli BL21 (DE3) using vector pET-28a-CtDFR and was purified. In vitro enzyme activity analysis, CtDFR could reduce three types of dihydroflavonols including dihydrokaempferol, dihydroquercetin, and dihydromyricetin to leucopelargonidin, leucocyanidin and leucodelphinidin. Subcellular localization analysis showed that CtDFR was mainly localized in the cytoplasm. These results demonstrate that CtDFR plays an important role in regulation of flower color in C. tubulosa and make a valuable contribution for the further investigation on the regulation mechanism of C. tubulosa (Schenk) Wight flower color.
ABSTRACT
1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.
ABSTRACT
Xanthine oxidoreductase (XOR), the key enzyme catalyzing purine to produce uric acid, including two subtypes, xanthine dehydrogenase (XDH) and xanthine oxidase (XO), respectively, in vivo. Usually, XDH and XO can transform to each other. In this study, based on the principle that the subtype XO or XDH uses different electron acceptors, the methods for the measuring the activities of bovine milk XOR (pure enzyme) and its subtypes were established. The optimal concentrations of substrate xanthine (50 μmol·L-1) and electron acceptor NAD+ (50 μmol·L-1), pH value (7.80) were investigated. The ranges of the XOR, XO, XDH activity which could be determined were 0.97-17.5 U·L-1, 1-9 U·L-1, and 66-1 191 mU·L-1, respectively. Furthermore, the methods for determining the activities of XOR and its subtypes in mouse liver were established. The preparation of liver samples, the optimal concentrations of xanthine (100 μmol·L-1) and NAD+ (100 μmol·L-1) were researched. And the activity ranges of XOR, XO and XDH in mouse liver which could be determined were 0.67-3.98, 0.19-1.08, and 0.52-3.55 U·gprot-1, respectively. With the methods above, the effects of classic XOR inhibitor allopurinal (Allo) on XOR, XO and XDH from both milk and mouse liver were determined. All animal experiments have been approved by the Animal Experimental Center, Institute of Materia Medica, Chinese Academy of Medical Science and Peking Union Medical College (00003346). This study established new methods for the determination of XOR and its subtypes activity in pure enzyme system and in mouse liver, respectively, which were accurate and convenient. It laid the experimental foundation for exploring the different pathophysiological effects of XOR in the body and developing new XOR inhibitors.
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ObjectiveTo screen the preparation technology of Baoyuan chewable tablets and to preliminarily elucidate its anti-fatigue effect and mechanism. MethodTaking encapsulation rate of volatile oil， extract rate and extraction rate of active ingredients as indexes， single factor test and orthogonal test were used to optimize the volatile oil inclusion， aqueous decoction and formulation molding processes of Baoyuan chewable tablets. ICR rats were randomly divided into the blank group， model group， Gaoshan Hongjingtian oral liquids group（6.01 mL·kg-1） and and Baoyuan chewable tablets low， medium， and high dose groups（2.1， 4.2， 8.4 g·kg-1）， 8 mice in each group， and were administered by gastric gavage at the corresponding dose once a day， the blank and model groups were given equal volume of saline for 15 d. After the last administration for 30 min， the mice were loaded with 5% of the body mass of lead at the tail and swam until exhaustion to establish the fatigue model， and the weighted swimming time of the mice in each group was recorded， meanwhile， the muscle tissues of the mice were sliced， stained by hematoxylin-eosin（HE） and subjected to pathological observation， and the levels of blood urea nitrogen（BUN）， lactic acid（LA）， liver glycogen（LG）， activities of lactate dehydrogenase（LDH） and creatine kinase（CK） in the serum were determined. ResultThe optimal inclusion process of cinnamon oil in Baoyuan chewable tablets was 10∶1 for β-cyclodextrin-volatile oil， and inclusion at 50 ℃ for 2 h with saturated aqueous solution method. The optimal water extraction process was to extract twice， adding 10 times of water to extract for 50 min for the first time， and adding 9 times of water to extract for 40 min for the second time. The ratio of the extract of Baoyuan chewable tablets with microcrystalline cellulose， maltodextrin， mannitol， citric acid， magnesium stearate was 63∶13∶8∶17∶17∶1∶1， the tablets were pressed by wet granulation， the each tablet weight was 1.2 g， and the hardness was 60-80 N. Compared with the model group， Baoyuan chewable tablets low， medium， and high dose groups could significantly prolong the exhaustion time of mice in weight bearing swimming（P<0.05， P<0.01）， and improve the exercise endurance of the body， and the results of HE staining showed that all dose groups of Baoyuan chewable tablets could significantly improve the muscle tissue damage caused by exercise， significantly reduce the levels of BUN， LA and the activities of LDH and CK in serum（P<0.01）， and significantly increase the content of LG（P<0.05， P<0.01）. ConclusionThe optimized preparation process of Baoyuan chewable tablets is stable and feasible， and the preparation can improve exercise endurance by increasing the LG level in liver tissue， and relieve muscle soreness by accelerating the removal of LA from the body， and reduce CK and LDH activities to exert anti-fatigue effects.
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@#Objective To express insulin-degrading enzyme(IDE)mutant T142A in prokaryotic cells and detect its activity.Methods According to the results of multi-sequence alignment and IDE substrate co-crystal structure,an active residue in β6-strand structure of IDE were predicted.The recombinant plasmid ppSUMO-T142A,with the site mutation of threonine 142 to alanine,was constructed by point mutation technique and expressed by E.coli prokaryotic expression system.After purification by nickel ion column affinity chromatography,ion exchange chromatography and gel filtration chromatography,the mutant T142A was obtained and determined for the activity by fluorescence method.Results IDE amino acid sequence is highly conserved among 16 species.T142 directly participates in substrate binding,interacts with substrate cleavage sites,and is close to important structures such as catalytic active sites and door-subdomains.The mutation of recombinant plasmid ppSUMO-T142A was proved to be correct by sequencing.The expressed fusion protein His-SUMO-T142A was mainly existed in soluble form in the supernatant at a concentration of 18 mg/mL,with a relative molecular mass of about 131 000;After three steps of purification,the purity of mutant T142A reached 86%.The maximum reaction rate(V_(max))of T142A catalytic degradation of fluorescent substrate V was 501.06 min~(-1) and the Michaelis constant(K_m) was 9.01μmol/L.Compared with wild-type IDE(V_(max) was 2 814.32 min~(-1),K_m was 11.93μmol/L),the activity of T142A decreased significantly.Conclusion The activity of IDE mutant T142A expressed in this study greatly decreases,while T142 is an important residue for IDE to play its enzymatic function,which provides an experimental basis for the development of new IDE activity regulatory molecules.
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@#Objective To express CamA(Clostridiodies difficile adenine methltransferase A,CamA)protein with methylase activity in prokaryotic expression system.Methods The gene sequence of CamA protein of the standard strain of C.difficile1870 was amplified by PCR,cloned into plasmid pGEX-4T-1-MBP,transformed into E.coli HB101 and induced by 1 mmol/L IPTG to express the recombinant target protein. After purification with Amylose Resin,CamA protein was digested by tobacco etch virus(TEV)protease,and verified for its methylase activity using MTase-Glo~(TM)Methyltransferase Assay in vitro.Results PCR sequencing showed that the cloned CamA gene sequence was correct,in which no base mutation occurred. The recombinant expression plasmid pGEX-4T-1-MBP-CamA was digested by EcoRⅠand BamHⅠ,which showed that 1 731 bp of CamA gene was connected to the vector plasmid. The relative molecular mass of the recombinant protein was about 108 800(with 1 MBP tag),and the purity was over 90% after purified with Amylose Resin. Under the condition of20 μmol/L DNA and 20 μmol/L SAM at room temperature for 30 min,0. 5 μg CamA produced SAH at a concentration of about 101. 60 nmol/L and the enzyme activity was(0. 339 ± 0. 027)U/mg.Conclusion The CamA protein of C.difficile has been successfully expressed and purified with methylase activity,which lays a foundation of further study on the function of CamA and its role in the occurrence,development and treatment of C.difficile infection.
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ObjectiveTo explore the forest type and soil environment suitable for Panax ginseng. MethodThe yield， quality， soil chemical properties， soil enzyme activity， and soil microbial metabolism of 9-year-old P. ginseng under different forests were investigated. ResultThe quality of P. ginseng was significantly different among forest types. To be specific， P. ginseng under the Quercus mongolica forest had the highest quality， with the total saponin content of 2.27% which was 51.89% higher than that in P. ginseng under Larix gmelinii forest. The yield of P. ginseng under Q. mongolica forest and L. gmelinii forest （30 g·m-2） was the highest， 62.5% higher than that under Betula platyphylla forest. The soil content of organic matter， Cu， and Zn， and activity of sucrase and urease under Q. mongolica forest were lower than those under other forest types. The utilization rate of D-galacturonic acid by soil microorganisms under Q. mongolica forest was higher than that under other forest types， but the utilization rate of L-phenylalanine was lower than that under other forest types. The utilization rate of 2-hydroxybenzoic acid by soil microorganisms of B. platyphylla forest was significantly lower than that under other forest types. There was a negative correlation between soil Zn and ginsenoside Rb1 and Rc， and between soil K and ginsenoside Rb2 and Rb3. Mn and Cu were positively correlated with most saponins. The results of redundancy analysis showed that the soil microorganisms using carbon sources of amino acids， esters， acids， and sugars were the main factors causing the differences in P. ginseng among different forest types. ConclusionThe yield and quality of P. ginseng under Q. mongolica forest were the best， followed by the forest with different tree species， and coming in last was the B. platyphylla forest. This study is expected to provide theoretical support for the improvement of P. ginseng yield and quality and the improvement of ecological planting technology.
ABSTRACT
This study aims to explore the resource utilization of used fungus-growing materials produced in the cultivation of Gastrodia elata. To be specific, based on the production practice, this study investigated the recycling mechanism of used fungus-growing materials of G. elata by Phallus inpudicus. To screen edible fungi with wide adaptability, this study examined the allelopathic effects of Armillaria mellea secretions on P. impudicus and 6 kinds of large edible fungi and the activities of enzymes related to degradation of the used fungus-growing materials of G. elata. The results showed that P. impudicus can effectively degrade cellulose, hemicellulose, and lignin in used fungus-growing materials of G. elata. The cellulase activity of A. mellea was significantly higher than that of P. impudicus, and the activities of lignin peroxidase, polyphenol oxidase, and xylanase of P. impudicus were significantly higher than those of A. mellea, which was the important reason why A. mellea and P. impudicus used different parts and components of the used fungus-growing materials to absorb carbon sources and develop ecological niche differences. The growth of P. impudicus was significantly inhibited on the used fungus-growing materials of G. elata. The secretions of A. mellea had allelopathic effects on P. impudicus and other edible fungi, and the allelopathic effects were related to the concentration of allelopathy substances. The screening result showed that the growth and development of L. edodes and A. auricular were not significantly affected by 30% of A. mellea liquid, indicating that they had high resistance to the allelopathy of A. mellea. The results showed that the activities of extracellular lignin peroxidase, polyphenol oxidase, and xylanase of the two edible fungi were similar to those of P. impudicus, and the cellulase activity was higher than that of P. impudicus. This experiment can be further verified by small-scale production tests.
Subject(s)
Agaricales , Ascomycota , Basidiomycota , Catechol Oxidase , Cellulases , GastrodiaABSTRACT
Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.
Subject(s)
Bromelains/isolation & purification , Enzyme Activation , Ammonium Sulfate , Peptide Hydrolases , Electrophoresis, Polyacrylamide GelABSTRACT
@#Trichinellosis is an important zoonotic parasitic disease worldwide and is principally caused by ingesting animal meat containing Trichinella infective larvae. Aspartyl aminopeptidase is an intracytoplasmic metalloproteinase that specifically hydrolyzes the N-terminus of polypeptides free of acidic amino acids (aspartic acid and glutamate), and plays an important role in the metabolism, growth and development of organisms. In this study, a novel T. spiralis aspartyl aminopeptidase (TsAAP) was cloned and expressed, and its biological properties and roles in worm growth and development were investigated. The results revealed that TsAAP transcription and expression in diverse T. spiralis stages were detected by RT-PCR and Western blotting, and primarily localized at cuticle, stichosome and intrauterine embryos of this nematode by immunofluorescence test. rTsAAP has the enzymatic activity of native AAP to hydrolyze the substrate H-Glu-pNA. There was a specific binding between rTsAAP and murine erythrocyte, and the binding site was localized in erythrocyte membrane proteins. Silencing of TsAAP gene by specific dsRNA significantly reduced the TsAAP expression, enzymatic activity, intestinal worm burdens and female fecundity. The results demonstrated that TsAAP participates in the growth, development and fecundity of T. spiralis and it might be a potential target molecule for anti-Trichinella vaccines.
ABSTRACT
With the rapid development of nuclear technology in the fields of industry and medicine, the possibility of people suffering from radiation damage has also increased. Radiation injury prevention and treatment drugs are the most effective and direct means for the treatment and protection of radiation injury, but the current radiation injury prevention and treatment drugs have limited effects. Due to the unique valence structure of cerium nanomaterials, it has a variety of enzymatic simulation activities and reproducibility. It shows superior oxidation resistance, powerful free radical scavenging function, and can protect cells from radiation damage. It can be used as an ideal it is a radioprotective agent, and is used in a variety of biological fields. A review of relevant literature shows that the antioxidant properties, high SOD mimicking activity, free radical scavenging ability and radiation resistance of ceria nanoparticles are derived from the mutual conversion of Ce3+/Ce4+ and the formation of oxygen vacancies. This article mainly introduces the basis of anti-radiation activity of ceria nanoparticles, radiation protection effects and the research progress of radiotherapy sensitization, and provides theoretical basis and reference for ceria nanoparticles in the field of radiation direction.
ABSTRACT
Ornithine decarboxylase (ODC) is a key enzyme in the biosynthetic pathway of polyamines and catalyzes the decarboxylation of ornithine to produce putrescine. Inhibition of ODC activity is a potential approach for the prevention and treatment of many diseases including cancer, as the expression levels and the activities of ODC in many abnormal cells and tumor cells are generally higher than those of normal cells. The discovery and evaluation of ODC inhibitors rely on the monitoring of the reaction processes catalyzed by ODC. There are several commonly used methods for analyzing the activity of ODC, such as measuring the yield of putrescine by high performance liquid chromatography, or quantifying the yield of isotope labelled carbon dioxide. However, the cumbersome operation and cost of these assays, as well as the difficulty to achieve high-throughput and real-time detection, hampered their applications. In this work, we optimized a real-time label-free method for analyzing the activity of ODC based on the macromolecule cucurbit[6]uril (CB6) and a fluorescent dye, DSMI (trans-4-[4-(dimethylamino) styryl]-1-methylpyridinium iodide). Finally, the optimized method was used to determine the activities of different ODC inhibitors with different inhibition mechanisms.
Subject(s)
Bridged-Ring Compounds , Imidazoles , Ornithine , Ornithine Decarboxylase , Ornithine Decarboxylase Inhibitors , PutrescineABSTRACT
To screen the available tomato pollution-safe cultivar varieties and reduce the potential food safety risks in Cd-polluted areas, the differences of Cd accumulation in different tomato (Solanum lycopersicum) varieties in southern China were studied by soil culture and hydroponic experiments. Firstly, the high and low accumulation varieties were selected from 25 tomato varieties under 2.94 mg/kg Cd stress by soil culture test, and then the responses of high and low accumulation tomato varieties to Cd stress were determined by hydroponic experiments. The results of soil culture test show that under 2.94 mg/kg Cd stress, there were significant differences in plant height, total biomass and yield among 25 tomato cultivars, and the Cd contents of fruits of all 25 tomato cultivars exceeded the highest limit value (0.05 mg/kg) of CAC (Codex alimentarius commission). Through cluster analysis, 7, 4 and 14 varieties accumulating relatively high, medium, and low concentrations of Cd in the fruits were screened, among which the highest, the lowest, and the average Cd contents in the fruits were 3.06 mg/kg DW, 1.47 mg/kg DW, and 2.21 mg/kg DW, respectively. The results of hydroponic experiment show that under the same concentration of Cd stress, Qiantangxuri F1, a high Cd accumulating variety, absorbed Cd faster, accumulated more Cd, used shorter oxidative stress response time and had stronger tolerance to Cd than Zhefen 3053, a low Cd accumulating variety. The typical high and low Cd accumulating varieties can provide a reference for agricultural production in heavy metal polluted areas and the development of molecular-assisted breeding methods of PSC. At present, cultivating low Cd accumulating PSC varieties and dynamic monitoring of Cd contents in tomato fruits are feasible methods in medium and light Cd-polluted areas.
Subject(s)
Antioxidants , Cadmium/toxicity , China , Solanum lycopersicum , Plant Breeding , Plant Roots/chemistry , Soil Pollutants/toxicityABSTRACT
L-asparaginase hydrolyzes L-asparagine to produce L-aspartic acid and ammonia. It is widely distributed in microorganisms, plants and serum of some rodents, and has important applications in the pharmaceutical and food industries. However, the poor thermal stability, low catalytic efficiency and low yield hampered the further application of L-asparaginase. In this paper, rational design and 5' untranslated region (5'UTR) design strategies were used to increase the specific enzyme activity and protein expression of L-asparaginase derived from Rhizomucor miehei (RmAsnase). The results showed that among the six mutants constructed through homology modeling combined with sequence alignment, the specific enzyme activity of the mutant A344E was 1.5 times higher than the wild type. Subsequently, a food-safe strain Bacillus subtilis 168/pMA5-A344E was constructed, and the UTR strategy was used for the construction of recombinant strain B. subtilis 168/pMA5 UTR-A344E. The enzyme activity of B. subtilis 168/pMA5 UTR-A344E was 7.2 times higher than that of B. subtilis 168/pMA5-A344E. The recombinant strain B. subtilis 168/pMA5 UTR-A344E was scaled up in 5 L fermenter, and the final yield of L-asparaginase was 489.1 U/mL, showing great potential for industrial application.
Subject(s)
Asparaginase/genetics , Bacillus subtilis/genetics , Industrial Microbiology , Protein Engineering , Rhizomucor/enzymology , Sequence AlignmentABSTRACT
Acetylcholinesterase (AChE) is a key enzyme used to detect organophosphorus pesticide residues by the enzyme inhibition method. An accidental discovery of a mutant strain with AChE activity was made in our laboratory during the process of AChE expression by