Butyrylcholinesterase(BChE;EC 3.1.1.8),an enzyme structurally related to acetylcholinesterase,is widely distributed in the human body.It plays a role in the detoxification of chemicals such as succinylcholine,a muscle...Butyrylcholinesterase(BChE;EC 3.1.1.8),an enzyme structurally related to acetylcholinesterase,is widely distributed in the human body.It plays a role in the detoxification of chemicals such as succinylcholine,a muscle relaxant used in anesthetic practice.BChE is well-known due to variant forms of the enzyme with little or no hydrolytic activity which exist in some endogamous communities and result in prolonged apnea following the administration of succinylcholine.Its other functions include the ability to hydrolyze acetylcholine,the cholinergic neurotransmitter in the brain,when its primary hydrolytic enzyme,acetylcholinesterase,is absent.To assess its potential roles,BChE was studied in relation to insulin resistance,type 2 diabetes mellitus,cognition,hepatic disorders,cardiovascular and cerebrovascular diseases,and inflammatory conditions.Individuals who lack the enzyme activity of BChE are otherwise healthy,until they are given drugs hydrolyzed by this enzyme.Therefore,BChE is a candidate for the study of loss-of-function mutations in humans.Studying individuals with variant forms of BChE can provide insights into whether they are protected against metabolic diseases.The potential utility of the enzyme as a biomarker for Alzheimer’s disease and the response to its drug treatment can also be assessed.展开更多
Lipolytic enzymes have attracted enormous attentions because of their ability in ester hydrolysis,ester synthesis,transesterification and other biochemical reactions.Bacteria are important sources of lipolytic enzymes...Lipolytic enzymes have attracted enormous attentions because of their ability in ester hydrolysis,ester synthesis,transesterification and other biochemical reactions.Bacteria are important sources of lipolytic enzymes applied in industry.Here,a novel lipolytic enzyme encoded by esterase gene est1347 was identified in Marinobacter flavimaris WLL162,and was purified and characterized.The lipolytic enzyme Est1347 consisted of 312 amino acid residues and a 21-amino-acids N-terminal signal peptide with a predicted molecular weight of 34.2 kDa.It belongs to family V of bacterial lipolytic enzymes based on the amino acid sequence homology analysis.Est1347 is a mesophilic and alkali-resistant enzyme with the highest activity at 45℃and pH 8.5;it is stable at temperatures below 50℃and pH 7.5–11.0.Est1347 showed a preference for middle-length chain substrate p-NPC10 and a wide range of other substrates.The Km,Vmax,Kcat and Kcat/Km values of Est1347 for p-NPC10 in pH 8.5 at 45℃were 0.9411 mmol L^(−1),1285μmol min^(−1)mg^(−1),698.91 s^(−1)and 743.65 s^(−1)(mmol L^(−1))^(−1),respectively.It is also tolerant to the metal ions,organic solvents and detergents.In conclusion,the esterase Est1347 laid a foundation for further study of bacterial lipolytic enzyme family V.展开更多
This study was aimed to analyze the effect of procyanidin B2(PC)and tannin acid(TA)on the activities of cholesterol esterase(CEase)and the inhibitory mechanisms of enzymatic activity.The interaction mechanisms were in...This study was aimed to analyze the effect of procyanidin B2(PC)and tannin acid(TA)on the activities of cholesterol esterase(CEase)and the inhibitory mechanisms of enzymatic activity.The interaction mechanisms were investigated by enzymatic kinetics,multi-spectroscopy methods,thermodynamics analysis,molecular docking,and dynamic simulations.PC and TA could bind with CEase and inhibit the activity of enzyme in a mixed-competitive manner and non-competitive manner,which was verified by molecular docking simulations and dynamics simulations.Also,PC and TA showed the synergistic inhibition with orlistat.Fluorescence,UVvis and the thermodynamic analysis revealed that the complexes were formed from CEase and inhibitors by noncovalent interaction.As revealed by the circular dichroism results,both PC and TA decreased enzymatic activities by altering the conformations of CEase.The inhibition of PC and TA on CEase might be one mechanism for its cholesterol-lowering effect.展开更多
Iron is an essential but excessively toxic nutrient. Although iron is rich in nature, the acquisition of iron is a challenge to life. Its solubility is very low because it is mostly in the form of oxidation or hydroxi...Iron is an essential but excessively toxic nutrient. Although iron is rich in nature, the acquisition of iron is a challenge to life. Its solubility is very low because it is mostly in the form of oxidation or hydroxide. In order to overcome this, microorganisms have evolved a variety of iron absorption pathways, the most important of which is the siderophore-dependent iron absorption pathway. Both bacteria and fungi require specific siderophore esterases to encourage the release of iron within the cell. A deeper understanding of siderophore esterases is crucial for the development of new antibacterial and antifungal diagnostic and therapeutic approaches. There have been many recent studies on anti-infectives via siderophore antibiotic couplers in which siderophore esterases have also played an important role, and in this review, we provide an overview of several of the more common iron carriers as well as siderophore esterases in terms of structure as well as function.展开更多
Background Ferulic acid esterase(FAE)-secreting Lactiplantibacillus plantarum A1(Lp A1)is a promising silage inoculant due to the FAE’s ability to alter the plant cell wall structure during ensiling,an action that is...Background Ferulic acid esterase(FAE)-secreting Lactiplantibacillus plantarum A1(Lp A1)is a promising silage inoculant due to the FAE’s ability to alter the plant cell wall structure during ensiling,an action that is expected to improve forage digestibility.However,little is known regarding the impacts of Lp A1 on rumen microbiota.Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa’s ensilage,in vitro rumen incubation and microbiota.Results Samples of fresh and ensiled alfalfa treated with(either Lp A1 or Lp MTD/1)or without additives(as control;CON)and ensiled for 30,60 and 90 d were used for fermentation quality,in vitro digestibility and batch culture study.Inoculants treated silage had lower(P<0.001)pH,acetic acid concentration and dry matter(DM)loss,but higher(P=0.001)lactic acid concentration than the CON during ensiling.Compared to the CON and Lp MTD/1,silage treated with Lp A1 had lower(P<0.001)aNDF,ADF,ADL,hemicellulose,and cellulose contents and higher(P<0.001)free ferulic acid concentration.Compared silage treated with Lp MTD/1,silage treated with Lp A1 had significantly(P<0.01)improved ruminal gas production and digestibility,which were equivalent to those of fresh alfalfa.Realtime PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen’s total bacteria,fungi,Ruminococcus albus and Ruminococcus flavefaciens,while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON.Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments.Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments.Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community.Conclusions Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility,and modulating the rumen fermentation to improve feed utilization.展开更多
A feruloyl esterase (FAE-C6) gene of 957 bp was isolated from rumen microbial metagenome, subcloned into pET32b vector, and expressed in </span><i><span style="font-family:Verdana;">Escheri...A feruloyl esterase (FAE-C6) gene of 957 bp was isolated from rumen microbial metagenome, subcloned into pET32b vector, and expressed in </span><i><span style="font-family:Verdana;">Escherichia</span></i><i><span style="font-family:Verdana;"> coli</span></i><span style="font-family:Verdana;">. The enzyme purified in active form, consisted of 319 amino acid residues, with a molecular weight of 43.7 based on SDS-PAGE. Homology modeling showed that the FAE contained the catalytic triad composed of Ser</span><sub><span style="font-family:Verdana;">154-</span></sub><span style="font-family:Verdana;">Asp</span><sub><span style="font-family:Verdana;">263</span></sub><span style="font-family:Verdana;">His</span><sub><span style="font-family:Verdana;">295</span></sub><span style="font-family:Verdana;"> and a classical Gly-X-Ser</span><sub><span style="font-family:Verdana;">154</span></sub><span style="font-family:Verdana;">-X-Gly nucleophile motif commonly found in esterases. The FAE-C6 was characterized using corn fiber as substrate. Its combining action with glycoside hydrolases (C, X, A) individually and in various combinations was studied with focus on the difference in</span></span><span style="font-family:Verdana;"> the</span><span style="font-family:Verdana;"> effects on FA and sugar release. Glycoside hydrolases with endo-xylanase included in the enzyme mixture showed significant impact on increasing the FA yield. For the release of sugar, FAE enhanced the yield in all hydrolase combinations moderately and endo-xylanase was not the key factor in the enzyme formulation.展开更多
A novel esterase Est C10 from B acillus sp. CX01 isolated from the deep sea of the Western Pacific Ocean and the functionalities of Est C10 was characterized. At present, the reports about the kinetic resolution of ra...A novel esterase Est C10 from B acillus sp. CX01 isolated from the deep sea of the Western Pacific Ocean and the functionalities of Est C10 was characterized. At present, the reports about the kinetic resolution of racemic methyl 2-chloropropionate were quite rare. So we developed deep-sea microbial esterase Est C10 as a novel biocatalyst in the kinetic resolution of racemic methyl 2-chloropropionate and generate( R)-methyl 2-chloropropionate with high enantiomeric excess(>99%) after the optimization of process parameters such as p H, temperature, organic co-solvents, surfactants, substrate concentration and reaction time. Notably, the optimal substrate concentration(80 mmol/L) of esterase Est C10 was higher than the kinetic resolution of another esterase, Est12-7(50 mmol/L). The novel microbial esterase Est C10 identified from the deep sea was a promising green biocatalyst in the generation of( R)-methyl 2-chloropropionate as well of many other valuable chiral chemicals in industry.展开更多
AIM:To evaluate the utility of measuring epigenetic alterations in pancreatic and biliary fluids in determining molecular markers for pancreatobiliary cancers.METHODS:DNA was extracted from undiluted pancreatic and bi...AIM:To evaluate the utility of measuring epigenetic alterations in pancreatic and biliary fluids in determining molecular markers for pancreatobiliary cancers.METHODS:DNA was extracted from undiluted pancreatic and biliary fluids.As a surrogate for a genomewide hypomethylation assay,levels of long interspersed nuclear element-1(LINE-1) methylation were analyzed using bisulfite pyrosequencing.CpG island hypermethylation of 10 tumor-associated genes,aryl-hydrocarbon receptor repressor,adenomatous polyposis coli,calcium channel,voltage dependent,T type α1G subunit,insulin-like growth factor 2,O-6-methyl-guanine-DNA methyltransferase,neurogenin 1,CDKN2A,runt-related transcription factor 3(RUNX3),secreted frizzled-related protein 1,and ubiquitin carboxyl-terminal esterase L1(UCHL1),was analyzed using MethyLight.To examine the role of CpG methylation and histone deacetylation in the silencing of UCHL1,human gallbladder carcinoma cell lines and pancreatic carcinoma cell lines were treated with 2 or 5 μmol/L 5-AZA-dC for 72 h or 100 nmol/L Trichostatin A for 24 h.After the treatment,UCHL1 expression was analyzed by real-time reverse transcription-polymerase chain reaction.RESULTS:Pancreatobiliary cancers exhibited significantly lower LINE-1 methylation levels in pancreatic and biliary fluids than did noncancerous pancreatobiliary disease(58.7% ± 4.3% vs 61.7% ± 2.2%,P = 0.027;53.8% ± 6.6% vs 57.5% ± 1.7%,P = 0.007);however,LINE-1 hypomethylation was more evident in pancreatic cancer tissues than in pancreatic fluids(45.4% ± 5.5% vs 58.7% ± 4.3%,P < 0.001).CpG island hypermethylation of tumor-associated genes was detected at various frequencies,but it was not correlated with LINE-1 hypomethylation.Hypermethylation of the UCHL1 gene was cancer-specific and most frequently detected in pancreatic(67%) or biliary(70%) fluids from patients with pancreatobiliary cancer.As a single marker,hypermethylation of the UCHL1 gene in pancreatic and biliary fluids was most useful for the detection of pancreatic and pancreatobiliary cancers,respectively(100% specificity).Hypermethylation of the UCHL1 and RUNX3 genes in pancreatic and biliary fluids was the most useful combined marker for pancreatic(87% sensitivity and 100% specificity) and pancreatobiliary(97% sensitivity and 100% specificity) cancers.Treatment with a demethylating agent,5-AZA-2'-deoxycytidine,restored UCHL1 expression in pancreatobiliary cancer cell lines.CONCLUSION:Our results suggest that hypermethylation of UCHL1 and RUNX3 in pancreatobiliary fluid might be useful for the diagnosis of pancreatobiliary cancers.展开更多
Genomic mining has identifi ed a novel microbial alkaline esterase from the Indian Ocean. This esterase was overexpressed in E. coli BL21(DE3) and further functionally characterized. Under optimal conditions(10 mmol/L...Genomic mining has identifi ed a novel microbial alkaline esterase from the Indian Ocean. This esterase was overexpressed in E. coli BL21(DE3) and further functionally characterized. Under optimal conditions(10 mmol/L substrate, p H 6.0, 2 h at 40 °C), this esterase can hydrolyze racemic methyl mandelate to( R)-methyl mandelate with very high optical purity(e. e. >99%) and yield(nearly 90%). Interestingly, the stereoselectivity of this esterase is opposite to that of two previously reported lipases that can generate( S)-methyl mandelate through the hydrolysis of racemic methyl mandelate. No organic solvents or other additives were required to optimize the optical purity and production of the fi nal chiral product(R)-methyl mandelate, which can potentially simplify the production procedure of( R)-methyl mandelate catalyzed by esterase.展开更多
With the aim of identifying novel thermostable esterases, comprehensive sequence databases and cloned fosmid libraries of metagenomes derived from an offshore oil reservoir on the Norwegian Continental Shelf were scre...With the aim of identifying novel thermostable esterases, comprehensive sequence databases and cloned fosmid libraries of metagenomes derived from an offshore oil reservoir on the Norwegian Continental Shelf were screened for enzyme candidates using both sequence-and function-based screening. From several candidates identified in both approaches, one enzyme discovered by the functional approach was verified as a novel esterase and subjected to a deeper characterization. The enzyme was successfully over-produced in Escherichia coli and was shown to be thermostable up to 90°C, with the highest esterase activity on short-chain ester substrates and with tolerance to solvents and metal ions. The fact that the thermostable enzyme was solely found by functional screening of the oil reservoir metagenomes illustrates the importance of this approach as a complement to purely sequence-based screening, in which the enzyme candidate was not detected. In addition, this example indicates the large potential of deep-sub-surface oil reservoir metagenomes as a source of novel, thermostable enzymes of potential relevance for industrial applications.展开更多
The GDSL esterase/lipase family contains many functional genes that perform important biological functions in growth and development, morphogenesis, seed oil synthesis, and defense responses in plants. The expression ...The GDSL esterase/lipase family contains many functional genes that perform important biological functions in growth and development, morphogenesis, seed oil synthesis, and defense responses in plants. The expression of GDSL esterase/lipase genes can respond to biotic and abiotic stresses. Although GDSL esterase/lipase family genes have been identified and studied in other plants, they have not been identified and their functions remain unclear in tomato. This study is the first to identify 80 GDSL esterase/lipase family genes in tomato, which were named SlGELP1–80. These genes were mapped to their positions on the chromosomes and their physical and chemical properties, gene structure, phylogenetic relationships, collinear relationships, and cis-acting elements were analyzed. The spatiotemporal expression characteristics of the Sl GELP genes in tomato were diverse. In addition, RNA-seq analysis indicated that the expression patterns of the SlGELP genes in tomato differed before and after inoculation with Stemphylium lycopersici. qRT-PCR was used to analyze the expression of five Sl GELP genes after treatments with S. lycopersici, salicylic acid and jasmonic acid. Finally, this study was the first to identify and analyze GDSL esterase/lipase family genes in tomato via bioinformatics approaches, and these findings provide new insights for improving the study of plant disease resistance.展开更多
This study was designed to assess total animal exposure to non-occupational but environmentally induced smoke through short-term landfill burning toxicity tests at the biochemical levels. Exposure to municipal land-fi...This study was designed to assess total animal exposure to non-occupational but environmentally induced smoke through short-term landfill burning toxicity tests at the biochemical levels. Exposure to municipal land-fill burning using rat model focused primarily on inhalation exposure. The environmental monitoring consisted of 60 days exposure to refuse burning by evaluating the level of protein concentrations, neurotransmitter enzyme acetylcholine esterase (AcHE), and total cholesterol in different tissues of Wistar rats. Protein concentrations tended to decrease in the brain, liver and kidney and slightly increased in the plasma while acetylcholine esterase decreased in brain and liver and increased in the kidney. The non-depletion in total cholesterol levels in the tissues tended to be due to active mobilization towards tissue metabolism. The data were sufficient to support risk assessment for human.展开更多
Juvenile hormone esterase(JHE) is a key enzyme for insects,playing an important role in the regulation of insect growth,development,diapause and reproduction.We identified a complete putative JHE of Cydia pomonella(Cp...Juvenile hormone esterase(JHE) is a key enzyme for insects,playing an important role in the regulation of insect growth,development,diapause and reproduction.We identified a complete putative JHE of Cydia pomonella(CpJHE-like) which is comprised of a 1 761 bp coding sequence(CDS) encoding 587 amino acid residues from the transcriptome data.The deduced protein sequence of CpJHE-like showed the highest identity of 60.44% with the Adoxophyes honmai JHE(AhJHE) and the minimal identity of 25.81% with Aedes aegypti JHE(AaJHE).CpJHE-like exhibited all the seven typical motifs of the functional JHEs and had the highly consistent tertiary structure with Manduca sexta JHE(MsJHE).Phylogenetic analysis showed that the CpJHE-like was close to two JHEs from the family Tortricidae.The CpJHE-like transcript level take a leap in the 3-day-old fifth instar larva,increased about 300-fold compared to the basal level.Tissue-specific expression profile showed that the CpJHE-like transcript was expressed mainly in the fat body.This study indicates that the CpJHE-like is the functional JHE,which may play vital roles in the development and reproduction of C.pomonella.展开更多
A continuous co-evolutionary arms-race between pathogens and their host plants promotes the development of pathogenic factors by microbes, including carbohydrate esterase(CE) genes to overcome the barriers in plant ce...A continuous co-evolutionary arms-race between pathogens and their host plants promotes the development of pathogenic factors by microbes, including carbohydrate esterase(CE) genes to overcome the barriers in plant cell walls. Identification of CEs is essential to facilitate their functional and evolutionary investigations; however, current methods may have a limit in detecting some conserved domains, and ignore evolutionary relationships of CEs, as well as do not distinguish CEs from proteases. Here, candidate CEs were annotated using conserved functional domains, and orthologous gene detection and phylogenetic relationships were used to identify new CEs in 16 oomycete genomes, excluding genes with protease domains. In our method, 41 new putative CEs were discovered comparing to current methods, including three CE4, 14 CE5, eight CE12, five CE13, and 11 CE14. We found that significantly more CEs were identified in Phytophthora than in Hyaloperonospora and Pythium, especially CE8, CE12, and CE13 that are putatively involved in pectin degradation. The abundance of these CEs in Phytophthora may be due to a high frequency of multiple-copy genes, supporting by the phylogenetic distribution of CE13 genes, which showed five units of Phytophthora CE13 gene clusters each displaying a species tree like topology, but without any gene from Hyaloperonospora or Pythium species. Additionally, diverse proteins associated with products of CE13 genes were identified in Phytophthora strains. Our analyses provide a highly effective method for CE discovery, complementing current methods, and have the potential to advance our understanding of function and evolution of CEs.展开更多
EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong diffe...EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong different stain recipes for Est of 3 kinds of fungi-Lentinus edodes. Pleurotus sapidus. Phellinus igriarius and 2 kinds of plants-Populus sp and Brassica chinensis. Of the four kinds of Est staining recipes tested.the recipe α-acetic acid-naphther showed the best effect.and followed by β-aceticacid-naphther, semicontent α-aceticacid-naphther and α+β-aceticacidnathpher.展开更多
Three diesters of exo- syn-meso-oxabicyclo (2, 2, 1 ) -hept- 5- ene- 2, 3- dicarboxylic acid and three tetraesters of tetrahydrofuran-2, 3, 4, 5-tetracarboxylic acid were synthesized and tested with enantioselective h...Three diesters of exo- syn-meso-oxabicyclo (2, 2, 1 ) -hept- 5- ene- 2, 3- dicarboxylic acid and three tetraesters of tetrahydrofuran-2, 3, 4, 5-tetracarboxylic acid were synthesized and tested with enantioselective hydrolysis catalyzed by pig liver esterase(PLE). The results of the PLEcatalyzed hydrolysis were discussed.展开更多
Germinating seeds of Tamarindus indica synthesizes various enzymes which are required for the degradation of seed reserves such as xyloglucans, fatty acid esters and proteins. Among these, esterases, belonging to a gr...Germinating seeds of Tamarindus indica synthesizes various enzymes which are required for the degradation of seed reserves such as xyloglucans, fatty acid esters and proteins. Among these, esterases, belonging to a group of hydrolytic enzymes catalyze the hydrolysis of various types of esters. They play an important role in cell expansion as well as detoxification of xenobiotics and many agrochemicals and insecticides. The esterases are extracted from the germinating tamarind seeds using 50 mM phosphate buffer, pH 7. The Km with α-naphthyl acetate as the substrate is 19.23 μM and the enzymes are optimally active at pH 7.0 to 7.5 and are stable between pH 5.0 to 9.0. The optimum temperature of esterase activity of tamarind seed is between 37?C - 50?C and is stable up to 40?C. The activity declined by 30% at 60?C and about 90% at 70?C. Highest esterase activity and specific activity are observed on the 21st day of germination. The polyacrylamide gel electrophoresis (PAGE) indicated the presence of nine isozymes of esterases. Band numbers 1, 5 and 6 are the major esterolytic bands present throughout the germination period while band numbers 2 & 3 are minor bands present only during the latter period of the germination. Based on substrate and inhibitor specificity in conjunction with electrophoresis, the esterases 1 to 8 have been classified as carboxylesterases sensitive to organophosphate inhibitor (OP) and PCMB (p-chloromercuribenzoate) while esterase 9 is classified as carboxylesterase sensitive to OP. These esterases are unaffected by carbamate inhibitor, eserine sulphate.展开更多
The present work was conducted to lower and alleviate the saline injury by using natural products in garlic extract application on growth, metabolites, protein pattern and esterase enzyme of wheat plants. This study w...The present work was conducted to lower and alleviate the saline injury by using natural products in garlic extract application on growth, metabolites, protein pattern and esterase enzyme of wheat plants. This study was conducted that wheat plant cv. Gimiza 11 response to osmotic stress effects and in general showed a variable response between different organs. The aerial parts of plants not only alleviated salinity injury but activated the fresh and dry matter productions. In root these parameters decreased as increasing salinity stress. Length of the shoots, roots and spikes run parallel with the previous results. Photosynthetic pigment enhanced markedly the increasing osmotic stress levels. The effect of garlic was reflected on the accumulation of soluble sugar and soluble protein in both roots and spikes, and a reduction of Na+ and an increase in K+ under garlic treatments were recorded. In the present study, staining intensity of protein bands of wheat plant was decreased as osmotic stress increased but the number of bands was increased up to -0.9 MPa, after that level a slight decrease was recorded (for control induction, 12 bands, -0.3 MPa, 16 bands, -0.6 MPa 14 bands, -0.9 14 bands, -1.2 MPa 11 bands and final 11 bands for -1.5 MPa). Induction protein bands for control plus garlic were 12 bands, for -0.3 MPa OSL plus garlic were 13 bands, for -0.6 MPa OSL plus garlic were 12 bands, for -0.9 MPa OSL plus garlic were 12 bands, for -1.2 MPa OSL were 8 bands and finally for -1.5 MPa OSL plus garlic were 9 bands. Electrophoresis studies of esterase showed wide variations in their intensities and densities among all treatments. There were 6 isozymes forms of esterase under OSL and with garlic but intensity was different. It seems that garlic extract was able to enhance the tolerance of the wheat plant to osmotic stress.展开更多
The total protein and esterase were isolated from the seeds of Red Kidney beans (Phaseolus vulgaris). The crude protein content was observed as 15%. The germination of seeds of red kidney bean has been carried out and...The total protein and esterase were isolated from the seeds of Red Kidney beans (Phaseolus vulgaris). The crude protein content was observed as 15%. The germination of seeds of red kidney bean has been carried out and change in the total protein content and esterase activity was monitored. The protein content was decreased from 15% (24 hours) to 8% (144 hours) during germination. The socked seeds and seedlings of all the days of germination exhibited esterase activity. Maximum ester hydrolyzing activity was observed on 6th day of germination whereas as lowest ester hydrolyzing activity was observed 2nd day germination. Native PAGE was carried out and esterase banding pattern for two different artificial substrates was studied. The esterase banding pattern in presence of 1-Napthyl acetate showed the presence of 4 esterolytic bands while 2 esterolytic bands were observed in presence of 2-Napthyl acetate.展开更多
文摘Butyrylcholinesterase(BChE;EC 3.1.1.8),an enzyme structurally related to acetylcholinesterase,is widely distributed in the human body.It plays a role in the detoxification of chemicals such as succinylcholine,a muscle relaxant used in anesthetic practice.BChE is well-known due to variant forms of the enzyme with little or no hydrolytic activity which exist in some endogamous communities and result in prolonged apnea following the administration of succinylcholine.Its other functions include the ability to hydrolyze acetylcholine,the cholinergic neurotransmitter in the brain,when its primary hydrolytic enzyme,acetylcholinesterase,is absent.To assess its potential roles,BChE was studied in relation to insulin resistance,type 2 diabetes mellitus,cognition,hepatic disorders,cardiovascular and cerebrovascular diseases,and inflammatory conditions.Individuals who lack the enzyme activity of BChE are otherwise healthy,until they are given drugs hydrolyzed by this enzyme.Therefore,BChE is a candidate for the study of loss-of-function mutations in humans.Studying individuals with variant forms of BChE can provide insights into whether they are protected against metabolic diseases.The potential utility of the enzyme as a biomarker for Alzheimer’s disease and the response to its drug treatment can also be assessed.
基金supported by the projects from the National Natural Science Foundation of China(No.42230411)the China Ocean Mineral Resources R and D Association(COMRA)Special Foundation(No.DY135-B2-10).
文摘Lipolytic enzymes have attracted enormous attentions because of their ability in ester hydrolysis,ester synthesis,transesterification and other biochemical reactions.Bacteria are important sources of lipolytic enzymes applied in industry.Here,a novel lipolytic enzyme encoded by esterase gene est1347 was identified in Marinobacter flavimaris WLL162,and was purified and characterized.The lipolytic enzyme Est1347 consisted of 312 amino acid residues and a 21-amino-acids N-terminal signal peptide with a predicted molecular weight of 34.2 kDa.It belongs to family V of bacterial lipolytic enzymes based on the amino acid sequence homology analysis.Est1347 is a mesophilic and alkali-resistant enzyme with the highest activity at 45℃and pH 8.5;it is stable at temperatures below 50℃and pH 7.5–11.0.Est1347 showed a preference for middle-length chain substrate p-NPC10 and a wide range of other substrates.The Km,Vmax,Kcat and Kcat/Km values of Est1347 for p-NPC10 in pH 8.5 at 45℃were 0.9411 mmol L^(−1),1285μmol min^(−1)mg^(−1),698.91 s^(−1)and 743.65 s^(−1)(mmol L^(−1))^(−1),respectively.It is also tolerant to the metal ions,organic solvents and detergents.In conclusion,the esterase Est1347 laid a foundation for further study of bacterial lipolytic enzyme family V.
基金supported by the National Basic Research Program of China(‘973’program,2013CB127106)。
文摘This study was aimed to analyze the effect of procyanidin B2(PC)and tannin acid(TA)on the activities of cholesterol esterase(CEase)and the inhibitory mechanisms of enzymatic activity.The interaction mechanisms were investigated by enzymatic kinetics,multi-spectroscopy methods,thermodynamics analysis,molecular docking,and dynamic simulations.PC and TA could bind with CEase and inhibit the activity of enzyme in a mixed-competitive manner and non-competitive manner,which was verified by molecular docking simulations and dynamics simulations.Also,PC and TA showed the synergistic inhibition with orlistat.Fluorescence,UVvis and the thermodynamic analysis revealed that the complexes were formed from CEase and inhibitors by noncovalent interaction.As revealed by the circular dichroism results,both PC and TA decreased enzymatic activities by altering the conformations of CEase.The inhibition of PC and TA on CEase might be one mechanism for its cholesterol-lowering effect.
文摘Iron is an essential but excessively toxic nutrient. Although iron is rich in nature, the acquisition of iron is a challenge to life. Its solubility is very low because it is mostly in the form of oxidation or hydroxide. In order to overcome this, microorganisms have evolved a variety of iron absorption pathways, the most important of which is the siderophore-dependent iron absorption pathway. Both bacteria and fungi require specific siderophore esterases to encourage the release of iron within the cell. A deeper understanding of siderophore esterases is crucial for the development of new antibacterial and antifungal diagnostic and therapeutic approaches. There have been many recent studies on anti-infectives via siderophore antibiotic couplers in which siderophore esterases have also played an important role, and in this review, we provide an overview of several of the more common iron carriers as well as siderophore esterases in terms of structure as well as function.
基金funded by National Natural Science Foundation of China(project no.31901390)China Postdoctoral Science Foundation(project no.2022M711451)Natural Science Foundation of Gansu Province,China(22JR5RA527)。
文摘Background Ferulic acid esterase(FAE)-secreting Lactiplantibacillus plantarum A1(Lp A1)is a promising silage inoculant due to the FAE’s ability to alter the plant cell wall structure during ensiling,an action that is expected to improve forage digestibility.However,little is known regarding the impacts of Lp A1 on rumen microbiota.Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa’s ensilage,in vitro rumen incubation and microbiota.Results Samples of fresh and ensiled alfalfa treated with(either Lp A1 or Lp MTD/1)or without additives(as control;CON)and ensiled for 30,60 and 90 d were used for fermentation quality,in vitro digestibility and batch culture study.Inoculants treated silage had lower(P<0.001)pH,acetic acid concentration and dry matter(DM)loss,but higher(P=0.001)lactic acid concentration than the CON during ensiling.Compared to the CON and Lp MTD/1,silage treated with Lp A1 had lower(P<0.001)aNDF,ADF,ADL,hemicellulose,and cellulose contents and higher(P<0.001)free ferulic acid concentration.Compared silage treated with Lp MTD/1,silage treated with Lp A1 had significantly(P<0.01)improved ruminal gas production and digestibility,which were equivalent to those of fresh alfalfa.Realtime PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen’s total bacteria,fungi,Ruminococcus albus and Ruminococcus flavefaciens,while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON.Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments.Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments.Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community.Conclusions Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility,and modulating the rumen fermentation to improve feed utilization.
文摘A feruloyl esterase (FAE-C6) gene of 957 bp was isolated from rumen microbial metagenome, subcloned into pET32b vector, and expressed in </span><i><span style="font-family:Verdana;">Escherichia</span></i><i><span style="font-family:Verdana;"> coli</span></i><span style="font-family:Verdana;">. The enzyme purified in active form, consisted of 319 amino acid residues, with a molecular weight of 43.7 based on SDS-PAGE. Homology modeling showed that the FAE contained the catalytic triad composed of Ser</span><sub><span style="font-family:Verdana;">154-</span></sub><span style="font-family:Verdana;">Asp</span><sub><span style="font-family:Verdana;">263</span></sub><span style="font-family:Verdana;">His</span><sub><span style="font-family:Verdana;">295</span></sub><span style="font-family:Verdana;"> and a classical Gly-X-Ser</span><sub><span style="font-family:Verdana;">154</span></sub><span style="font-family:Verdana;">-X-Gly nucleophile motif commonly found in esterases. The FAE-C6 was characterized using corn fiber as substrate. Its combining action with glycoside hydrolases (C, X, A) individually and in various combinations was studied with focus on the difference in</span></span><span style="font-family:Verdana;"> the</span><span style="font-family:Verdana;"> effects on FA and sugar release. Glycoside hydrolases with endo-xylanase included in the enzyme mixture showed significant impact on increasing the FA yield. For the release of sugar, FAE enhanced the yield in all hydrolase combinations moderately and endo-xylanase was not the key factor in the enzyme formulation.
基金Supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA11030404)the Guangzhou Science and Technology Plan Projects(No.201510010012)the National Natural Science Foundation of China(No.21302199)
文摘A novel esterase Est C10 from B acillus sp. CX01 isolated from the deep sea of the Western Pacific Ocean and the functionalities of Est C10 was characterized. At present, the reports about the kinetic resolution of racemic methyl 2-chloropropionate were quite rare. So we developed deep-sea microbial esterase Est C10 as a novel biocatalyst in the kinetic resolution of racemic methyl 2-chloropropionate and generate( R)-methyl 2-chloropropionate with high enantiomeric excess(>99%) after the optimization of process parameters such as p H, temperature, organic co-solvents, surfactants, substrate concentration and reaction time. Notably, the optimal substrate concentration(80 mmol/L) of esterase Est C10 was higher than the kinetic resolution of another esterase, Est12-7(50 mmol/L). The novel microbial esterase Est C10 identified from the deep sea was a promising green biocatalyst in the generation of( R)-methyl 2-chloropropionate as well of many other valuable chiral chemicals in industry.
基金Supported by Grants-in-Aid for Scientific Research from the Ministry of Education,Culture,Sports,Science and Technology of Japan (to Yamamoto H and Shinomura Y)
文摘AIM:To evaluate the utility of measuring epigenetic alterations in pancreatic and biliary fluids in determining molecular markers for pancreatobiliary cancers.METHODS:DNA was extracted from undiluted pancreatic and biliary fluids.As a surrogate for a genomewide hypomethylation assay,levels of long interspersed nuclear element-1(LINE-1) methylation were analyzed using bisulfite pyrosequencing.CpG island hypermethylation of 10 tumor-associated genes,aryl-hydrocarbon receptor repressor,adenomatous polyposis coli,calcium channel,voltage dependent,T type α1G subunit,insulin-like growth factor 2,O-6-methyl-guanine-DNA methyltransferase,neurogenin 1,CDKN2A,runt-related transcription factor 3(RUNX3),secreted frizzled-related protein 1,and ubiquitin carboxyl-terminal esterase L1(UCHL1),was analyzed using MethyLight.To examine the role of CpG methylation and histone deacetylation in the silencing of UCHL1,human gallbladder carcinoma cell lines and pancreatic carcinoma cell lines were treated with 2 or 5 μmol/L 5-AZA-dC for 72 h or 100 nmol/L Trichostatin A for 24 h.After the treatment,UCHL1 expression was analyzed by real-time reverse transcription-polymerase chain reaction.RESULTS:Pancreatobiliary cancers exhibited significantly lower LINE-1 methylation levels in pancreatic and biliary fluids than did noncancerous pancreatobiliary disease(58.7% ± 4.3% vs 61.7% ± 2.2%,P = 0.027;53.8% ± 6.6% vs 57.5% ± 1.7%,P = 0.007);however,LINE-1 hypomethylation was more evident in pancreatic cancer tissues than in pancreatic fluids(45.4% ± 5.5% vs 58.7% ± 4.3%,P < 0.001).CpG island hypermethylation of tumor-associated genes was detected at various frequencies,but it was not correlated with LINE-1 hypomethylation.Hypermethylation of the UCHL1 gene was cancer-specific and most frequently detected in pancreatic(67%) or biliary(70%) fluids from patients with pancreatobiliary cancer.As a single marker,hypermethylation of the UCHL1 gene in pancreatic and biliary fluids was most useful for the detection of pancreatic and pancreatobiliary cancers,respectively(100% specificity).Hypermethylation of the UCHL1 and RUNX3 genes in pancreatic and biliary fluids was the most useful combined marker for pancreatic(87% sensitivity and 100% specificity) and pancreatobiliary(97% sensitivity and 100% specificity) cancers.Treatment with a demethylating agent,5-AZA-2'-deoxycytidine,restored UCHL1 expression in pancreatobiliary cancer cell lines.CONCLUSION:Our results suggest that hypermethylation of UCHL1 and RUNX3 in pancreatobiliary fluid might be useful for the diagnosis of pancreatobiliary cancers.
基金Supported by the National Natural Science Foundation of China(No.21302199)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA11030404)+1 种基金the Project of“Engineering HighPerformance Microorganisms for Advanced Bio-Based Manufacturing”from the Chinese Academy of Sciences(No.KGZD-EW-606)the Guangzhou Science and Technology Plan Projects(No.201510010012)
文摘Genomic mining has identifi ed a novel microbial alkaline esterase from the Indian Ocean. This esterase was overexpressed in E. coli BL21(DE3) and further functionally characterized. Under optimal conditions(10 mmol/L substrate, p H 6.0, 2 h at 40 °C), this esterase can hydrolyze racemic methyl mandelate to( R)-methyl mandelate with very high optical purity(e. e. >99%) and yield(nearly 90%). Interestingly, the stereoselectivity of this esterase is opposite to that of two previously reported lipases that can generate( S)-methyl mandelate through the hydrolysis of racemic methyl mandelate. No organic solvents or other additives were required to optimize the optical purity and production of the fi nal chiral product(R)-methyl mandelate, which can potentially simplify the production procedure of( R)-methyl mandelate catalyzed by esterase.
文摘With the aim of identifying novel thermostable esterases, comprehensive sequence databases and cloned fosmid libraries of metagenomes derived from an offshore oil reservoir on the Norwegian Continental Shelf were screened for enzyme candidates using both sequence-and function-based screening. From several candidates identified in both approaches, one enzyme discovered by the functional approach was verified as a novel esterase and subjected to a deeper characterization. The enzyme was successfully over-produced in Escherichia coli and was shown to be thermostable up to 90°C, with the highest esterase activity on short-chain ester substrates and with tolerance to solvents and metal ions. The fact that the thermostable enzyme was solely found by functional screening of the oil reservoir metagenomes illustrates the importance of this approach as a complement to purely sequence-based screening, in which the enzyme candidate was not detected. In addition, this example indicates the large potential of deep-sub-surface oil reservoir metagenomes as a source of novel, thermostable enzymes of potential relevance for industrial applications.
基金supported by the“Bai Qian Wan”Project of Heilongjiang Province,China(2019ZX16B02)the National Natural Science Foundation of China(32002059)+1 种基金the Heilongjiang Natural Science Foundation of China(LH2020C10)the Fellowship of China Postdoctoral Science Foundation(2020M681068)。
文摘The GDSL esterase/lipase family contains many functional genes that perform important biological functions in growth and development, morphogenesis, seed oil synthesis, and defense responses in plants. The expression of GDSL esterase/lipase genes can respond to biotic and abiotic stresses. Although GDSL esterase/lipase family genes have been identified and studied in other plants, they have not been identified and their functions remain unclear in tomato. This study is the first to identify 80 GDSL esterase/lipase family genes in tomato, which were named SlGELP1–80. These genes were mapped to their positions on the chromosomes and their physical and chemical properties, gene structure, phylogenetic relationships, collinear relationships, and cis-acting elements were analyzed. The spatiotemporal expression characteristics of the Sl GELP genes in tomato were diverse. In addition, RNA-seq analysis indicated that the expression patterns of the SlGELP genes in tomato differed before and after inoculation with Stemphylium lycopersici. qRT-PCR was used to analyze the expression of five Sl GELP genes after treatments with S. lycopersici, salicylic acid and jasmonic acid. Finally, this study was the first to identify and analyze GDSL esterase/lipase family genes in tomato via bioinformatics approaches, and these findings provide new insights for improving the study of plant disease resistance.
文摘This study was designed to assess total animal exposure to non-occupational but environmentally induced smoke through short-term landfill burning toxicity tests at the biochemical levels. Exposure to municipal land-fill burning using rat model focused primarily on inhalation exposure. The environmental monitoring consisted of 60 days exposure to refuse burning by evaluating the level of protein concentrations, neurotransmitter enzyme acetylcholine esterase (AcHE), and total cholesterol in different tissues of Wistar rats. Protein concentrations tended to decrease in the brain, liver and kidney and slightly increased in the plasma while acetylcholine esterase decreased in brain and liver and increased in the kidney. The non-depletion in total cholesterol levels in the tissues tended to be due to active mobilization towards tissue metabolism. The data were sufficient to support risk assessment for human.
基金supported by the National Key Research and Development Program of China(2016YFC1201200,2017YFC1200600 and 2016YFC1200602)the Basic Research on Science and Technology Project of Shenzhen,China(JCYJ20160530191934833)the Dapeng New District Industrial Development Special Fund of Shenzhen,China(KY20180215)
文摘Juvenile hormone esterase(JHE) is a key enzyme for insects,playing an important role in the regulation of insect growth,development,diapause and reproduction.We identified a complete putative JHE of Cydia pomonella(CpJHE-like) which is comprised of a 1 761 bp coding sequence(CDS) encoding 587 amino acid residues from the transcriptome data.The deduced protein sequence of CpJHE-like showed the highest identity of 60.44% with the Adoxophyes honmai JHE(AhJHE) and the minimal identity of 25.81% with Aedes aegypti JHE(AaJHE).CpJHE-like exhibited all the seven typical motifs of the functional JHEs and had the highly consistent tertiary structure with Manduca sexta JHE(MsJHE).Phylogenetic analysis showed that the CpJHE-like was close to two JHEs from the family Tortricidae.The CpJHE-like transcript level take a leap in the 3-day-old fifth instar larva,increased about 300-fold compared to the basal level.Tissue-specific expression profile showed that the CpJHE-like transcript was expressed mainly in the fat body.This study indicates that the CpJHE-like is the functional JHE,which may play vital roles in the development and reproduction of C.pomonella.
基金supported by the Special Fund for Agro-scientific Research in the Public Interest,China(201303018)the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-ASTIP-IVFCAAS)the emarked fund for the China Agriculture Research System(CARS-25-B-01)
文摘A continuous co-evolutionary arms-race between pathogens and their host plants promotes the development of pathogenic factors by microbes, including carbohydrate esterase(CE) genes to overcome the barriers in plant cell walls. Identification of CEs is essential to facilitate their functional and evolutionary investigations; however, current methods may have a limit in detecting some conserved domains, and ignore evolutionary relationships of CEs, as well as do not distinguish CEs from proteases. Here, candidate CEs were annotated using conserved functional domains, and orthologous gene detection and phylogenetic relationships were used to identify new CEs in 16 oomycete genomes, excluding genes with protease domains. In our method, 41 new putative CEs were discovered comparing to current methods, including three CE4, 14 CE5, eight CE12, five CE13, and 11 CE14. We found that significantly more CEs were identified in Phytophthora than in Hyaloperonospora and Pythium, especially CE8, CE12, and CE13 that are putatively involved in pectin degradation. The abundance of these CEs in Phytophthora may be due to a high frequency of multiple-copy genes, supporting by the phylogenetic distribution of CE13 genes, which showed five units of Phytophthora CE13 gene clusters each displaying a species tree like topology, but without any gene from Hyaloperonospora or Pythium species. Additionally, diverse proteins associated with products of CE13 genes were identified in Phytophthora strains. Our analyses provide a highly effective method for CE discovery, complementing current methods, and have the potential to advance our understanding of function and evolution of CEs.
文摘EST isozymes are one of the frequently used biochemical markers in genetic analysis of fungi. and the staining is an important process in electrophoresis analysis of studying Est.The effects were compared ainong different stain recipes for Est of 3 kinds of fungi-Lentinus edodes. Pleurotus sapidus. Phellinus igriarius and 2 kinds of plants-Populus sp and Brassica chinensis. Of the four kinds of Est staining recipes tested.the recipe α-acetic acid-naphther showed the best effect.and followed by β-aceticacid-naphther, semicontent α-aceticacid-naphther and α+β-aceticacidnathpher.
基金Supported by the National Natural Science Foundation of China and Schweizerischer National fonds Zur Forderunyder wissenschaft
文摘Three diesters of exo- syn-meso-oxabicyclo (2, 2, 1 ) -hept- 5- ene- 2, 3- dicarboxylic acid and three tetraesters of tetrahydrofuran-2, 3, 4, 5-tetracarboxylic acid were synthesized and tested with enantioselective hydrolysis catalyzed by pig liver esterase(PLE). The results of the PLEcatalyzed hydrolysis were discussed.
文摘Germinating seeds of Tamarindus indica synthesizes various enzymes which are required for the degradation of seed reserves such as xyloglucans, fatty acid esters and proteins. Among these, esterases, belonging to a group of hydrolytic enzymes catalyze the hydrolysis of various types of esters. They play an important role in cell expansion as well as detoxification of xenobiotics and many agrochemicals and insecticides. The esterases are extracted from the germinating tamarind seeds using 50 mM phosphate buffer, pH 7. The Km with α-naphthyl acetate as the substrate is 19.23 μM and the enzymes are optimally active at pH 7.0 to 7.5 and are stable between pH 5.0 to 9.0. The optimum temperature of esterase activity of tamarind seed is between 37?C - 50?C and is stable up to 40?C. The activity declined by 30% at 60?C and about 90% at 70?C. Highest esterase activity and specific activity are observed on the 21st day of germination. The polyacrylamide gel electrophoresis (PAGE) indicated the presence of nine isozymes of esterases. Band numbers 1, 5 and 6 are the major esterolytic bands present throughout the germination period while band numbers 2 & 3 are minor bands present only during the latter period of the germination. Based on substrate and inhibitor specificity in conjunction with electrophoresis, the esterases 1 to 8 have been classified as carboxylesterases sensitive to organophosphate inhibitor (OP) and PCMB (p-chloromercuribenzoate) while esterase 9 is classified as carboxylesterase sensitive to OP. These esterases are unaffected by carbamate inhibitor, eserine sulphate.
文摘The present work was conducted to lower and alleviate the saline injury by using natural products in garlic extract application on growth, metabolites, protein pattern and esterase enzyme of wheat plants. This study was conducted that wheat plant cv. Gimiza 11 response to osmotic stress effects and in general showed a variable response between different organs. The aerial parts of plants not only alleviated salinity injury but activated the fresh and dry matter productions. In root these parameters decreased as increasing salinity stress. Length of the shoots, roots and spikes run parallel with the previous results. Photosynthetic pigment enhanced markedly the increasing osmotic stress levels. The effect of garlic was reflected on the accumulation of soluble sugar and soluble protein in both roots and spikes, and a reduction of Na+ and an increase in K+ under garlic treatments were recorded. In the present study, staining intensity of protein bands of wheat plant was decreased as osmotic stress increased but the number of bands was increased up to -0.9 MPa, after that level a slight decrease was recorded (for control induction, 12 bands, -0.3 MPa, 16 bands, -0.6 MPa 14 bands, -0.9 14 bands, -1.2 MPa 11 bands and final 11 bands for -1.5 MPa). Induction protein bands for control plus garlic were 12 bands, for -0.3 MPa OSL plus garlic were 13 bands, for -0.6 MPa OSL plus garlic were 12 bands, for -0.9 MPa OSL plus garlic were 12 bands, for -1.2 MPa OSL were 8 bands and finally for -1.5 MPa OSL plus garlic were 9 bands. Electrophoresis studies of esterase showed wide variations in their intensities and densities among all treatments. There were 6 isozymes forms of esterase under OSL and with garlic but intensity was different. It seems that garlic extract was able to enhance the tolerance of the wheat plant to osmotic stress.
文摘The total protein and esterase were isolated from the seeds of Red Kidney beans (Phaseolus vulgaris). The crude protein content was observed as 15%. The germination of seeds of red kidney bean has been carried out and change in the total protein content and esterase activity was monitored. The protein content was decreased from 15% (24 hours) to 8% (144 hours) during germination. The socked seeds and seedlings of all the days of germination exhibited esterase activity. Maximum ester hydrolyzing activity was observed on 6th day of germination whereas as lowest ester hydrolyzing activity was observed 2nd day germination. Native PAGE was carried out and esterase banding pattern for two different artificial substrates was studied. The esterase banding pattern in presence of 1-Napthyl acetate showed the presence of 4 esterolytic bands while 2 esterolytic bands were observed in presence of 2-Napthyl acetate.
