Glutamate dehydrogenase (GDH)-synthesized RNA, a nongenetic code-based RNA is suitable for unraveling the structural constraints imposed on the regulation (transcription, translation, siRNA etc.) of metabolism by genetic code. GDH-synthesized RNAs have been induced in whole plants to knock out target mRNA populations thereby producing plant phenotypes that are allergen-free; enriched in fatty acids, essential amino acids, shikimic acid, resveratrol etc. Methods applied hereunder for investigating the structural properties of GDH-synthesized RNA included purification of GDH isoenzymes, synthesis of RNA by the isoenzymes, reverse transcription of the RNA to cDNA, sequencing of the cDNA, computation of the G+C-contents, profiling the stability through PCR amplification compared with genetic code-based DNA; and biochemical characterization of the RNAs synthesized by individual hexameric isoenzymes of GDH. Single product bands resulted from the PCR amplification of the cDNAs of GDH-synthesized RNA, whereas several bands resulted from the amplification of genetic code-based DNA. The cDNAs have wide G+C-contents (35% to 59%), whereas genetic code-based DNA has narrower G+C-contents (50% to 60%). The GDH β6 homo-hexameric isoenzyme synthesized the A+U-rich RNAs, whereas the a6, and α6 homo-hexameric isoenzymes synthesized the G+C-rich RNAs. Therefore, the RNA synthesized by GDH is different from genetic code-based RNAs. In vitro chemical reactions revealed that GDH-synthesized RNA degraded total RNA to lower molecular weight products. Therefore, GDH-synthesized RNA is RNA enzyme. Dismantling of the structural constraints imposed on RNA by genetic code liberated RNA to become an enzyme with specificity to degrade unwanted transcripts. The RNA enzyme activity of GDH-synthesized RNA is ubiquitous in cells; it is readily induced by treatment of plants with mineral nutrients etc. and may simplify experimental approaches in plant enzymology and molecular biology research projects.
{"title":"Structural Properties of the RNA Synthesized by Glutamate Dehydrogenase for the Degradation of Total RNA","authors":"G. Osuji, Paul M Johnson","doi":"10.4236/AER.2018.63004","DOIUrl":"https://doi.org/10.4236/AER.2018.63004","url":null,"abstract":"Glutamate dehydrogenase (GDH)-synthesized RNA, a nongenetic code-based RNA is suitable for unraveling the structural constraints imposed on the regulation (transcription, translation, siRNA etc.) of metabolism by genetic code. GDH-synthesized RNAs have been induced in whole plants to knock out target mRNA populations thereby producing plant phenotypes that are allergen-free; enriched in fatty acids, essential amino acids, shikimic acid, resveratrol etc. Methods applied hereunder for investigating the structural properties of GDH-synthesized RNA included purification of GDH isoenzymes, synthesis of RNA by the isoenzymes, reverse transcription of the RNA to cDNA, sequencing of the cDNA, computation of the G+C-contents, profiling the stability through PCR amplification compared with genetic code-based DNA; and biochemical characterization of the RNAs synthesized by individual hexameric isoenzymes of GDH. Single product bands resulted from the PCR amplification of the cDNAs of GDH-synthesized RNA, whereas several bands resulted from the amplification of genetic code-based DNA. The cDNAs have wide G+C-contents (35% to 59%), whereas genetic code-based DNA has narrower G+C-contents (50% to 60%). The GDH β6 homo-hexameric isoenzyme synthesized the A+U-rich RNAs, whereas the a6, and α6 homo-hexameric isoenzymes synthesized the G+C-rich RNAs. Therefore, the RNA synthesized by GDH is different from genetic code-based RNAs. In vitro chemical reactions revealed that GDH-synthesized RNA degraded total RNA to lower molecular weight products. Therefore, GDH-synthesized RNA is RNA enzyme. Dismantling of the structural constraints imposed on RNA by genetic code liberated RNA to become an enzyme with specificity to degrade unwanted transcripts. The RNA enzyme activity of GDH-synthesized RNA is ubiquitous in cells; it is readily induced by treatment of plants with mineral nutrients etc. and may simplify experimental approaches in plant enzymology and molecular biology research projects.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"06 1","pages":"29-52"},"PeriodicalIF":0.0,"publicationDate":"2018-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43775030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Combinatorial chemistry has been a focus of research activity in modern drug discovery and biotechnology. It is a concept by which a vast library of molecular diversity is synthesized and screened for target properties. This report is to illustrate the application of enzyme technology using the concept of combinatorial chemistry as a novel approach for the bioconversion of plant fibers. Citrus pectin was subjected to combinatorial enzyme digestion to create libraries of pectic oligosaccharides with diverse structural variants. Repeated cycles of fractionation and screening resulted in the isolation and identification of an active oligoGalA species with antimicrobial activity.
{"title":"Combinatorial Enzyme Approach to Produce Oligosaccharides of Diverse Structures for Functional Screen","authors":"D. Wong, S. Feng, W. Orts","doi":"10.4236/AER.2018.62002","DOIUrl":"https://doi.org/10.4236/AER.2018.62002","url":null,"abstract":"Combinatorial chemistry has been a focus of research activity in modern drug discovery and biotechnology. It is a concept by which a vast library of molecular diversity is synthesized and screened for target properties. This report is to illustrate the application of enzyme technology using the concept of combinatorial chemistry as a novel approach for the bioconversion of plant fibers. Citrus pectin was subjected to combinatorial enzyme digestion to create libraries of pectic oligosaccharides with diverse structural variants. Repeated cycles of fractionation and screening resulted in the isolation and identification of an active oligoGalA species with antimicrobial activity.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"06 1","pages":"11-20"},"PeriodicalIF":0.0,"publicationDate":"2018-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42786888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yan Chen, G. Pu, B. Lian, X. Pei, Guifang Huang, Qifeng Wang, Yanna Lv
Fungi are the key agents in litter decomposition in forest ecosystems. However, the specific roles of the interactions between different fungal species during litter decomposition process are unclear. To evaluate the interactions, two fungi strains with significantly different morphs were isolated from the soils of Quercus acutissima forest and Pinus massoniana forest, and inoculated in the litter powder of Quercus acutissima leaves and Pinus massoniana needles with grown separately and in coexistence equally through a microcosm experiment. The enzyme activities were determined as a proxy for microbial activities. The results showed that the degradative enzymes involved in litter decomposition showed varying dynamics pattern during the incubation period. The interactions between the two fungi strains are synergism, and benefit to each other according to enzyme activities, suggesting that a fungi strain growth was accelerated by the presence of other fungi strain during litter decomposition process. However, the interactions of the two fungi strains were bilateral antagonism inoculated in the litter powder of Quercus acutissima leaves according to cellobiohydrolase activities. The synergism, despite bilateral antagonism in an exceptional case, may be an important factor controlling the fungal colonization and growth on litter substrate. The results implied that more fungal species may accelerate litter decomposition rates due to their mutual cooperation.
{"title":"Interactions between Two Fungi Strains during Litter Decomposition through a Microcosm Experiment: Different Degradative Enzyme Activities","authors":"Yan Chen, G. Pu, B. Lian, X. Pei, Guifang Huang, Qifeng Wang, Yanna Lv","doi":"10.4236/AER.2018.61001","DOIUrl":"https://doi.org/10.4236/AER.2018.61001","url":null,"abstract":"Fungi are the key agents in litter decomposition in forest ecosystems. However, the specific roles of the interactions between different fungal species during litter decomposition process are unclear. To evaluate the interactions, two fungi strains with significantly different morphs were isolated from the soils of Quercus acutissima forest and Pinus massoniana forest, and inoculated in the litter powder of Quercus acutissima leaves and Pinus massoniana needles with grown separately and in coexistence equally through a microcosm experiment. The enzyme activities were determined as a proxy for microbial activities. The results showed that the degradative enzymes involved in litter decomposition showed varying dynamics pattern during the incubation period. The interactions between the two fungi strains are synergism, and benefit to each other according to enzyme activities, suggesting that a fungi strain growth was accelerated by the presence of other fungi strain during litter decomposition process. However, the interactions of the two fungi strains were bilateral antagonism inoculated in the litter powder of Quercus acutissima leaves according to cellobiohydrolase activities. The synergism, despite bilateral antagonism in an exceptional case, may be an important factor controlling the fungal colonization and growth on litter substrate. The results implied that more fungal species may accelerate litter decomposition rates due to their mutual cooperation.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"6 1","pages":"1-9"},"PeriodicalIF":0.0,"publicationDate":"2018-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47747521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Andrew, N. S. Emmanuel, D. C. Ejiogu, C. N. Chima, F. M. Yahuza, E. C. Umeh, P. P. Mshelia
Isoniazid induced hepatotoxicity is a major concern in patients taking anti tuberculosis treatment and prophylaxis. It can result in elevated serum liver enzymes and hepatic failure. The aim of the study was to evaluate the phytochemicals and ameliorative effects of aqueous extracts of Brysocarpus coccineus on serum liver enzymes in isoniazid (INH) induced hepatotoxicity in adult male Wistar rats. Thirty six (36) adult male Wistar rats were divided into six groups of six rats each and were treated orally for 30 days as follows: Group I: 1 ml/kg of distilled water; group II: Isoniazid (27 mg/kg); group III: Isoniazid (27 mg/kg) + Livolin forte (20 mg/kg); group IV: Isoniazid (27 mg/kg) + B. coccineus (200 mg/kg); group V: Isoniazid (27 mg/kg) + B. coccineus (400 mg/kg); group VI: Isoniazid (27 mg/kg) + B. coccineus (800 mg/kg). At the end of the experiments, the Wistar rats were sacrificed and sera obtained for liver enzymes assay, whereas the liver tissue was also harvested and used for histological studies. Tanins, saponins, alkaloids and flavonoids were quantitatively present at 2.29%, 18.05%, 23.24% and 18.99%, respectively. There was an increase in the serum AST and ALT in the isoniazid treated group, which was reversed by livolin forte and the aqueous extracts at a dose of 200 mg/kg, however the extracts increased the serum levels of AST and ALT at higher doses, which was however not significant (p > 0.05) when compared to the controls. There was evidence of a reduction in hepatocytes damage in the extract treated groups when compared to the Isoniazid untreated group. In conclusion, aqueous extracts of B. coccineus shows hepatoprotective effects at 200 mg/kg in isoniazid hepatotoxicity in adult male Wistar rats.
{"title":"Evaluation of Quantitative Phytochemicals, Liver Enzymes and Histological Changes in Isoniazid Induced Hepatotoxicity in Adult Male Wistar Rats Treated with Aqueous Extracts of Brysocarpus coccineus","authors":"K. Andrew, N. S. Emmanuel, D. C. Ejiogu, C. N. Chima, F. M. Yahuza, E. C. Umeh, P. P. Mshelia","doi":"10.4236/AER.2017.53004","DOIUrl":"https://doi.org/10.4236/AER.2017.53004","url":null,"abstract":"Isoniazid induced hepatotoxicity is a major concern in patients taking anti tuberculosis treatment and prophylaxis. It can result in elevated serum liver enzymes and hepatic failure. The aim of the study was to evaluate the phytochemicals and ameliorative effects of aqueous extracts of Brysocarpus coccineus on serum liver enzymes in isoniazid (INH) induced hepatotoxicity in adult male Wistar rats. Thirty six (36) adult male Wistar rats were divided into six groups of six rats each and were treated orally for 30 days as follows: Group I: 1 ml/kg of distilled water; group II: Isoniazid (27 mg/kg); group III: Isoniazid (27 mg/kg) + Livolin forte (20 mg/kg); group IV: Isoniazid (27 mg/kg) + B. coccineus (200 mg/kg); group V: Isoniazid (27 mg/kg) + B. coccineus (400 mg/kg); group VI: Isoniazid (27 mg/kg) + B. coccineus (800 mg/kg). At the end of the experiments, the Wistar rats were sacrificed and sera obtained for liver enzymes assay, whereas the liver tissue was also harvested and used for histological studies. Tanins, saponins, alkaloids and flavonoids were quantitatively present at 2.29%, 18.05%, 23.24% and 18.99%, respectively. There was an increase in the serum AST and ALT in the isoniazid treated group, which was reversed by livolin forte and the aqueous extracts at a dose of 200 mg/kg, however the extracts increased the serum levels of AST and ALT at higher doses, which was however not significant (p > 0.05) when compared to the controls. There was evidence of a reduction in hepatocytes damage in the extract treated groups when compared to the Isoniazid untreated group. In conclusion, aqueous extracts of B. coccineus shows hepatoprotective effects at 200 mg/kg in isoniazid hepatotoxicity in adult male Wistar rats.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"05 1","pages":"33-44"},"PeriodicalIF":0.0,"publicationDate":"2017-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48515098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Tezuka, A. Higashino, M. Akiba, Takashi Nakamura
Assays of stress enzymes related to active oxygen species were performed by using an in vitro preparation from the liver of a monkey (Japanese Macaque). Ge-132, an organic germanium compound, viz. poly-trans-[(2-carboxyethyl) germasesquioxane] [(GeCH2CH2COOH)2O3]n, suppressed the activities of NADH-dependent oxidase and NADPH-dependent oxidase [NAD(P)H-OD] and xanthine oxidase (XOD) as superoxide-forming enzymes, while promoting the activities of superoxide dismutase (SOD) as a superoxide-scavenging enzyme and catalase (CAT) as an enzyme responsible for degradation of hydrogen peroxide (H2O2). The evidence suggests that the levels of active oxygen species such as and H2O2 would be reduced by Ge-132. The possible connection between Ge-132 and activities of stress enzymes is discussed on the basis of these results.
{"title":"Organogermanium (Ge-132) Suppresses Activities of Stress Enzymes Responsible for Active Oxygen Species in Monkey Liver Preparation","authors":"T. Tezuka, A. Higashino, M. Akiba, Takashi Nakamura","doi":"10.4236/AER.2017.52002","DOIUrl":"https://doi.org/10.4236/AER.2017.52002","url":null,"abstract":"Assays of stress enzymes related to active oxygen species were performed by using an in vitro preparation from the liver of a monkey (Japanese Macaque). Ge-132, an organic germanium compound, viz. poly-trans-[(2-carboxyethyl) germasesquioxane] [(GeCH2CH2COOH)2O3]n, suppressed the activities of NADH-dependent oxidase and NADPH-dependent oxidase [NAD(P)H-OD] and xanthine oxidase (XOD) as superoxide-forming enzymes, while promoting the activities of superoxide dismutase (SOD) as a superoxide-scavenging enzyme and catalase (CAT) as an enzyme responsible for degradation of hydrogen peroxide (H2O2). The evidence suggests that the levels of active oxygen species such as and H2O2 would be reduced by Ge-132. The possible connection between Ge-132 and activities of stress enzymes is discussed on the basis of these results.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"05 1","pages":"13-23"},"PeriodicalIF":0.0,"publicationDate":"2017-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46413693","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
I. Malgwi, Y. Tanko, M. Kawu, E. Eze, H. Salami, N. S. Emmanuel, A. Mohammed
In mammals, lactation is the most energetically demanding period of a female’s reproductive life. This study was designed to evaluate the effect of fermented Soya bean and Vitamin C supplement on lipid peroxidation and antioxidant enzymes in lactating albino rats. Thirty five (35) adult female rats were used for this study. At parturition, the animals were randomly divided into five groups of five (5) rats each. Except group four (4) that was subdivided into three (3) sub groups of five animals each (n = 5). Treatment was carried out as follows: Group I: (Normal control) was given normal feed and distilled water, orally (1 ml/kg), Group II: metoclopramide (5 mg/kg), Group III: 100 mg/kg of Vitamin C. The three (3) sub groups under group four (4) received 10%, 20% and 40% soya bean, respectively, Group V: was co-administered with 20% soya bean supplement and Vitamin C (100 mg/kg). Treatment was done for the period of ten (10) days at 06:00 h daily. Although there was an increase in serum MDA concentrations in all the treated groups compared to the control, lipid peroxidation was however significantly higher (P < 0.05) in the metoclopramide group relative to the soya bean supplemented groups. This study has shown that supplementation with soya bean induces a mild antioxidant effect by increasing serum level of superoxide dismutase. There was however a significant decrease in serum SOD in the 10% SB group compared to the control. There was a significant difference in serum catalase activity in the group treated with METCL (46.20 ± 1.53), SB 10% (44.00 ± 1.14) and SB 20% (45.20 ± 1.28) compared to the control (52.00 ± 0.71) (P < 0.05). Serum level of glutathione peroxidase GPx showed a significant difference in the group treated with VIT C, SB 10% and SB 20% compared to the control (P < 0.05).
{"title":"Lipid Peroxidation and Antioxidant Enzymes Evaluation in Lactating Female Albino Rats Following Supplementation with Fermented Soya Bean and Vitamin C","authors":"I. Malgwi, Y. Tanko, M. Kawu, E. Eze, H. Salami, N. S. Emmanuel, A. Mohammed","doi":"10.4236/AER.2017.52003","DOIUrl":"https://doi.org/10.4236/AER.2017.52003","url":null,"abstract":"In mammals, lactation is the most energetically demanding period of a female’s reproductive life. This study was designed to evaluate the effect of fermented Soya bean and Vitamin C supplement on lipid peroxidation and antioxidant enzymes in lactating albino rats. Thirty five (35) adult female rats were used for this study. At parturition, the animals were randomly divided into five groups of five (5) rats each. Except group four (4) that was subdivided into three (3) sub groups of five animals each (n = 5). Treatment was carried out as follows: Group I: (Normal control) was given normal feed and distilled water, orally (1 ml/kg), Group II: metoclopramide (5 mg/kg), Group III: 100 mg/kg of Vitamin C. The three (3) sub groups under group four (4) received 10%, 20% and 40% soya bean, respectively, Group V: was co-administered with 20% soya bean supplement and Vitamin C (100 mg/kg). Treatment was done for the period of ten (10) days at 06:00 h daily. Although there was an increase in serum MDA concentrations in all the treated groups compared to the control, lipid peroxidation was however significantly higher (P < 0.05) in the metoclopramide group relative to the soya bean supplemented groups. This study has shown that supplementation with soya bean induces a mild antioxidant effect by increasing serum level of superoxide dismutase. There was however a significant decrease in serum SOD in the 10% SB group compared to the control. There was a significant difference in serum catalase activity in the group treated with METCL (46.20 ± 1.53), SB 10% (44.00 ± 1.14) and SB 20% (45.20 ± 1.28) compared to the control (52.00 ± 0.71) (P < 0.05). Serum level of glutathione peroxidase GPx showed a significant difference in the group treated with VIT C, SB 10% and SB 20% compared to the control (P < 0.05).","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"05 1","pages":"24-31"},"PeriodicalIF":0.0,"publicationDate":"2017-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47966770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Acetylcholinesterase (AChE) is an important enzyme responsible for the cleavage of acetylcholine. Studies of the activity of this enzyme use an artificial substrate, acetylthiocholine, because a product of its catalysis, thiocholine, readily generates a light absorbing product upon reaction with Elman’s reagent 5,5’-dithiobis-(2-nitrobenzoic acid (DTNB). The hydrolysis of acetylcholine cannot be assayed with this method. The isothermal titration calorimetry can assay the hydrolysis of both substrates, without requiring additional reagents other than the enzyme and the substrate. To compare kinetic values obtained in the hydrolysis of acetylcholine (ACh) and acetylthiocholine (ATCh), with carbaryl acting as inhibitor, a calorimetric technique was used to evaluate kinetic properties of the two reactions. This method can show the hydrolysis of both substrates by the heat exchange that occurs during catalysis. In addition, it allowed the assessment of the AChE inhibition by carbaryl, a common insecticide. The results show a similarity between values obtained with both substrates, which are slightly higher for acetylcholine, the enzyme natural substrate. Enzymatic parameters values from ATCh and ACh were similar to each other and inhibitory constants using carbaryl were also similar, displaying that any approach to ACh is feasible using ATCh. The results obtained from ITC show the precision achieved by the calorimetric method.
{"title":"Microcalorimetric Study of Acetylcholine and Acetylthiocholine Hydrolysis by Acetylcholinesterase","authors":"P. Neves, Eliane Novato Silva, P. Beirão","doi":"10.4236/AER.2017.51001","DOIUrl":"https://doi.org/10.4236/AER.2017.51001","url":null,"abstract":"Acetylcholinesterase (AChE) is an important enzyme responsible for the cleavage of acetylcholine. Studies of the activity of this enzyme use an artificial substrate, acetylthiocholine, because a product of its catalysis, thiocholine, readily generates a light absorbing product upon reaction with Elman’s reagent 5,5’-dithiobis-(2-nitrobenzoic acid (DTNB). The hydrolysis of acetylcholine cannot be assayed with this method. The isothermal titration calorimetry can assay the hydrolysis of both substrates, without requiring additional reagents other than the enzyme and the substrate. To compare kinetic values obtained in the hydrolysis of acetylcholine (ACh) and acetylthiocholine (ATCh), with carbaryl acting as inhibitor, a calorimetric technique was used to evaluate kinetic properties of the two reactions. This method can show the hydrolysis of both substrates by the heat exchange that occurs during catalysis. In addition, it allowed the assessment of the AChE inhibition by carbaryl, a common insecticide. The results show a similarity between values obtained with both substrates, which are slightly higher for acetylcholine, the enzyme natural substrate. Enzymatic parameters values from ATCh and ACh were similar to each other and inhibitory constants using carbaryl were also similar, displaying that any approach to ACh is feasible using ATCh. The results obtained from ITC show the precision achieved by the calorimetric method.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"05 1","pages":"1-12"},"PeriodicalIF":0.0,"publicationDate":"2017-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46181961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Andreeva Na, L. Trilisenko, M. Eldarov, T. Kulakovskaya
The Saccharomyces cerevisiae polyphosphatase PPN1 (uniprot/Q04119) degrades inorganic polyphosphates both by cleaving Pi from the chain end and by fragmenting long-chain polymers into shorter ones. In this study, we have found a new activity of this protein: it releases phosphate from dATP. The dATP phosphohydrolase activity of pure PPN1 was ~7-fold lower compared to the exopolyphosphatase activity. This activity was strongly stimulated by Co2+ ions, as well as by ammonium ions, and inhibited by heparin and pyrophosphate similar to the exopolyphosphatase activity of PPN1. The Km value for dATP was 0.88 ± 0.14 mM. The dATP phosphohydrolase activity in the cells of PPN1-overexpressing yeast strain was several-fold higher than that in the parent strain. The other exopolyphosphatase of S. cerevisiae, PPX1, did not split Pi from dATP.
{"title":"Polyphosphatase PPN1 of Saccharomyces cerevisiae Is a Deoxyadenosine Triphosphate Phosphohydrolase","authors":"Andreeva Na, L. Trilisenko, M. Eldarov, T. Kulakovskaya","doi":"10.4236/AER.2016.44013","DOIUrl":"https://doi.org/10.4236/AER.2016.44013","url":null,"abstract":"The Saccharomyces cerevisiae polyphosphatase PPN1 (uniprot/Q04119) degrades inorganic polyphosphates both by cleaving Pi from the chain end and by fragmenting long-chain polymers into shorter ones. In this study, we have found a new activity of this protein: it releases phosphate from dATP. The dATP phosphohydrolase activity of pure PPN1 was ~7-fold lower compared to the exopolyphosphatase activity. This activity was strongly stimulated by Co2+ ions, as well as by ammonium ions, and inhibited by heparin and pyrophosphate similar to the exopolyphosphatase activity of PPN1. The Km value for dATP was 0.88 ± 0.14 mM. The dATP phosphohydrolase activity in the cells of PPN1-overexpressing yeast strain was several-fold higher than that in the parent strain. The other exopolyphosphatase of S. cerevisiae, PPX1, did not split Pi from dATP.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"04 1","pages":"144-151"},"PeriodicalIF":0.0,"publicationDate":"2016-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70485707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Castor oil is the source of numerous products and is the only commercial source of the fatty acid ricinoleate, 12-hydroxy-oleate. Hydrogenated castor oil is similarly useful as the source of 12-hydroxy-stearic acid, best known as a component of lithium grease. Mono- and diacylglycerols are derived from castor oil and are useful in development of lubricants and emulsifiers for cosmetics, pharmaceutical and food use. Acylglycerols derived from hydrogenated castor oil may be similarly useful, albeit with different physical and chemical properties. We have evaluated the use of immobilized lipases to generate acylglycerols, using organic solvents to modulate the action of lipase to produce mono- and diacylglycerols, using tri-(12-hydroxy stearoyl)-glycerol as a model for hydrogenated castor oil. The presence of an alkylated oxygen in the solvent appears to be an important factor in supporting lipase activity, with diisopropyl ether providing the best yield of di-(12-hydroxy stearoyl)-glycerol.
{"title":"Lipase Catalyzed Methanolysis of Tri-(12-Hydroxy Stearoyl)-Glycerol in Organic Solvents","authors":"S. Kang, T. Mckeon","doi":"10.4236/aer.2016.44014","DOIUrl":"https://doi.org/10.4236/aer.2016.44014","url":null,"abstract":"Castor oil is the source of numerous products and is the only commercial source of the fatty acid ricinoleate, 12-hydroxy-oleate. Hydrogenated castor oil is similarly useful as the source of 12-hydroxy-stearic acid, best known as a component of lithium grease. Mono- and diacylglycerols are derived from castor oil and are useful in development of lubricants and emulsifiers for cosmetics, pharmaceutical and food use. Acylglycerols derived from hydrogenated castor oil may be similarly useful, albeit with different physical and chemical properties. We have evaluated the use of immobilized lipases to generate acylglycerols, using organic solvents to modulate the action of lipase to produce mono- and diacylglycerols, using tri-(12-hydroxy stearoyl)-glycerol as a model for hydrogenated castor oil. The presence of an alkylated oxygen in the solvent appears to be an important factor in supporting lipase activity, with diisopropyl ether providing the best yield of di-(12-hydroxy stearoyl)-glycerol.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"04 1","pages":"152-157"},"PeriodicalIF":0.0,"publicationDate":"2016-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70485751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Osmar Soares da Silva, R. L. C. Oliveira, C. Souza-Motta, A. Porto, T. S. Porto
This study reports the protease production from Aspergillus tamarii using agroindustrial residues as substrate for solid-state fermentation (SSF) and biochemical characterization. The highest protease production was obtained using wheat bran as substrate at 72 h fermentation with maximum proteolytic activity of 401.42 U/mL, collagenase of 243.0 U/mL and keratinase of 19.1 U/mL. The protease exhibited KM = 18.7 mg/mL and Vmax = 28.5 mg/mL/min. The optimal pH was 8.0 and stable in a wide pH range (5.0 - 11.0) during 24 h. The optimum temperature was 40°C. The proteolytic activity was inhibited by Cu2+ (33.98%) and Hg2+ (22.69%). The enzyme was also inhibited by PMSF (65.11%), indicating that is a Serine Protease. These properties suggest that alkaline protease from A. tamarii URM4634 is suitable for application in food industries and leather processing. Additionally, the present findings opened new vistas in the utilization of wheat bran and other effective agroindustrial wastes as substrates for SSF.
{"title":"Novel Protease from Aspergillus tamarii URM4634: Production and Characterization Using Inexpensive Agroindustrial Substrates by Solid-State Fermentation","authors":"Osmar Soares da Silva, R. L. C. Oliveira, C. Souza-Motta, A. Porto, T. S. Porto","doi":"10.4236/AER.2016.44012","DOIUrl":"https://doi.org/10.4236/AER.2016.44012","url":null,"abstract":"This study reports the protease production from Aspergillus tamarii using agroindustrial residues as substrate for solid-state fermentation (SSF) and biochemical characterization. The highest protease production was obtained using wheat bran as substrate at 72 h fermentation with maximum proteolytic activity of 401.42 U/mL, collagenase of 243.0 U/mL and keratinase of 19.1 U/mL. The protease exhibited KM = 18.7 mg/mL and Vmax = 28.5 mg/mL/min. The optimal pH was 8.0 and stable in a wide pH range (5.0 - 11.0) during 24 h. The optimum temperature was 40°C. The proteolytic activity was inhibited by Cu2+ (33.98%) and Hg2+ (22.69%). The enzyme was also inhibited by PMSF (65.11%), indicating that is a Serine Protease. These properties suggest that alkaline protease from A. tamarii URM4634 is suitable for application in food industries and leather processing. Additionally, the present findings opened new vistas in the utilization of wheat bran and other effective agroindustrial wastes as substrates for SSF.","PeriodicalId":65616,"journal":{"name":"酶研究进展(英文)","volume":"04 1","pages":"125-143"},"PeriodicalIF":0.0,"publicationDate":"2016-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70485697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：