Abstract:
:Horseradish peroxidase (HRP) was successfully purified from horseradish roots by a two-stage reverse-micellar extraction from the dialyzed aqueous extract. The anionic surfactant AOT dissolved in isooctane was used to produce the reverse-micellar phases. The narrow pH range at which HRP solubilization occurred was exploited to remove most of the contaminant proteins in the first forward extraction. In the second extraction stage, HRP was selectively solubilized and concentrated by using a volume ratio of 10 between the aqueous and organic phases. The HRP final specific activity was 86 guaiacol U mg-1, obtained with a purification factor of 80 and yield of 46%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed two overlapping bands, with HRP corresponding to that at 43.8 kDa. Image analysis on isoelectric focusing (IEF) gels showed that the HRP was 80% pure. Ion exchange liquid chromatography showed that most of the specific activity was due to the basic isoenzyme with pI 8.5, which comprises 33.5% of the product. There were high HRP losses as a precipitate at the interface when direct reverse-micellar extraction was attempted from the crude extract. It is believed that the hydrophobic environment near the haem group of the HRP basic isoenzyme favors complex formation with the surfactant, and that this is promoted at higher protein concentrations.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Regalado C,Asenjo JA,Pyle DLdoi
10.1016/0141-0229(95)00031-3subject
Has Abstractpub_date
1996-04-01 00:00:00pages
332-9issue
5eissn
0141-0229issn
1879-0909pii
0141022995000313journal_volume
18pub_type
杂志文章abstract::To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzy...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2019.109447
更新日期:2020-02-01 00:00:00
abstract::A straightforward synthesis of gold nanoparticles (AuNps) is achieved by novel probiotic Lactobacillus kimchicus DCY51T isolated from Korean kimchi via an intracellular membrane-bound mechanism. The bioreduction of HAuCl4 into AuNps was verified by ultraviolet-visible spectrophotometry at ∼540nm. AuNps were spherical ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.08.018
更新日期:2016-12-01 00:00:00
abstract::Solid-liquid enzyme reactions constitute important processes in biochemical industries. The isomerization of d-glucose to d-fructose, using the immobilized glucose isomerase (Sweetzyme T), as a typical example of solid-liquid catalyzed reactions has been carried out in one stage and multi-stage novel type of impinging...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2013.11.006
更新日期:2014-02-05 00:00:00
abstract::The kinetic and thermodynamic parameters of wheat β-amylase (WBA) were characterized and various additives were evaluated for enhancing its activity and thermostability. WBA activity was examined by neocuproine method using soluble starch as substrate. The Michaelis constant (K(m)) and molecular activity (k(cat)) were...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2012.07.004
更新日期:2012-10-10 00:00:00
abstract::l-ribose and d-tagatose are biochemically synthesized using sugar isomerases. The l-arabinose isomerase gene from Shigella flexneri (Sf-AI) was cloned and expressed in Escherichia coli BL-21. Sf-AI was applied for the bioproduction of d-tagatose from d-galactose. l-ribose synthesis was performed by two step isomerizat...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.11.001
更新日期:2017-02-01 00:00:00
abstract::Isolates from gardening waste compost and 38 culture collection microbes were grown on agar plates at pH 4.0 with the cutinase model substrate polycaprolactone as a carbon source. The strains showing polycaprolactone hydrolysis were cultivated in liquid at acidic pH and the cultivations were monitored by assaying the ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2013.01.005
更新日期:2013-04-10 00:00:00
abstract::Phospholipase C (PLC) activity secreted by bacteria as a virulence factor is commonly detected by use of the artificial substrate p-nitrophenylphosphorylcholine (p-NPPC). We examined several commercially available enzymes (phosphodiesterases, phosphomonoesterases, phospholipase A, lipase, protease) for their hydrolyti...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(99)00190-8
更新日期:2000-03-01 00:00:00
abstract::In this work, a biosensor using a glassy carbon electrode modified with gold nanoparticles (AuNPs) and tyrosinase (Tyr) within a dihexadecylphosphate film is proposed. Cystamine and glutaraldehyde crosslinking agents were used as a support for Tyr immobilization. The proposed biosensor was characterized by scanning el...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.12.004
更新日期:2016-03-01 00:00:00
abstract::Two acidic β-glucosidases (βGI and βGII) from the brown rot fungus Fomitopsis palustris were purified to homogeneity by several chromatographic steps. βGI and βGII had molecular weights of 130 and 213 kDa, respectively, and exhibited optimum activity at pH 2.5 and 55°C. The K(m) values of βGI and βGII for p-nitropheny...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2010.12.012
更新日期:2011-04-07 00:00:00
abstract::The effect of intense pulsed light (IPL) irradiation on Chromobacterium viscosum lipase was investigated with a primary focus on catalytic activity and molecular structure. During IPL irradiation, lipase activity decreased significantly with increasing pulse fluence (Fp) and exposure time (te). IPL-induced deactivatio...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2019.02.001
更新日期:2019-05-01 00:00:00
abstract::In the previous study, glutathione production was elevated by adding precursor amino acids and adenosine triphosphate (ATP) in high cell density cultivation of Candida utilis. Furthermore, in the present research, glutathione production was further improved by optimizing sodium dedecyl sulfate (SDS) addition coupled w...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2010.02.007
更新日期:2010-05-05 00:00:00
abstract::Monod kinetic parameters provide information required for kinetic analysis of anaerobic oxidation of methane coupled to denitrification (AOM-D). This information is critical for engineering AOM-D processes in wastewater treatment facilities. We first experimentally determined Monod kinetic parameters for an AOM-D enri...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.05.005
更新日期:2017-09-01 00:00:00
abstract::Production of laccase activity by nine strains of Chalara paradoxa isolated from olive mill wastewater disposal ponds were studied. Enzyme extracts obtained from cultured broths by adsorption on hydroxyapatite showed a single band of laccase activity on ABTS after polyacrylamide gel electrophoresis (PAGE). They showed...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(99)00197-0
更新日期:2000-04-01 00:00:00
abstract::Mycobacterium tuberculosis acetohydroxyacid synthase (M. tuberculosis AHAS) has been proposed to bean essential target for novel herbicide- and chemical-based antibacterial agents. Therefore, here we investigated the roles of multiple conserved herbicide-binding site residues (R318, A146, Q148, M512, and V513) in M. t...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.06.009
更新日期:2015-10-01 00:00:00
abstract::Malolactic fermentation (MLF) is a secondary bioconversion developed in some wines involving malic acid decarboxylation. The induction of MLF in wine by cultures of free and immobilized Oenococcus oeni cells was investigated. This work reports on the effect of surface charges in the immobilization material, a recently...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00339-2
更新日期:2001-03-08 00:00:00
abstract::Microorganism selection is critical to deglycosylation in soybean fermentation for producing beneficial phytochemicals. This study investigated isoflavone bioconversion in soybean extract inoculated with Lactobacillus plantarum K2-12 and Lactobacillus curvatus JD0-31 exhibiting different enzyme activities. L. plantaru...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2019.109394
更新日期:2020-01-01 00:00:00
abstract::Various recombinant proteins can be produced by the yeast Saccharomyces cerevisiae cell factories; therefore, efficient recombinant protein production techniques are desirable. In this study, to establish an efficient recombinant protein production technique in S. cerevisiae, the secretory production of recombinant pr...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2018.10.014
更新日期:2019-02-01 00:00:00
abstract::We here describe a unique β-D-glucosidase (BGL; Blon_0625) derived from Bifidobacterium longum subsp. infantis ATCC 15697. The Blon_0625 gene was expressed by recombinant Escherichia coli. Purified recombinant Blon_0625 retains hydrolyzing activity against both p-nitrophenyl-β-D-glucopyranoside (pNPG; 17.3±0.24Umg(-1)...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.10.001
更新日期:2015-01-01 00:00:00
abstract::Ergosterol, like cholesterol, has many beneficial physiological activities, and because it has no adverse clinical problem with cholesterol, it is a substance that can be used as a biomaterial in the food and cosmetics industry. However, ergosterol has low oil solubility and is easily crystallized, which is problemati...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2020.109581
更新日期:2020-09-01 00:00:00
abstract::The different physical and chemical properties of lignin might have various effects on the enzymatic hydrolysis of lignocellulosic substrates. In this study, the influence of lignin on enzymatic digestibility of cellulose was assessed. Addition of 20% (4g/L) isolated enzymatic lignin (lignin 2 and 3) and kraft lignin ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.03.001
更新日期:2017-06-01 00:00:00
abstract::Different lipases have been immobilized on octyl agarose beads at 1mg/g and at maximum loading, via physical interfacial activation versus the octyl layer on the support. The stability of the preparations was analyzed. Most biocatalysts had the expected result: the apparent stability increased using the highly loaded ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.12.002
更新日期:2017-03-01 00:00:00
abstract::Fremyella diplosiphon is a freshwater cyanobacterium that has great potential as a biofuel agent due to its ability to grow in low light intensity and acclimation to different wavelengths. To enhance its halotolerance for growth in 35gL-1 sodium chloride (NaCl), plasmids harboring hemolysin B (hlyB) and malate dehydro...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.04.009
更新日期:2017-08-01 00:00:00
abstract::Electricity production from carbon monoxide (CO) is demonstrated in a single chamber microbial fuel cell (MFC) with a CoTMPP-based air cathode. The MFC was inoculated with anaerobic sludge and continuously sparged with CO as a sole carbon source. Volumetric power output was maximized at a CO flow rate of 4.8LLR(-1)d(-...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2010.02.010
更新日期:2010-05-05 00:00:00
abstract::The filamentous fungal strains Aspergillus niger I-1472 and Pycnoporus cinnabarinus MUCL39533, previously selected for the bioconversion of ferulic acid to vanillic acid and vanillin respectively, were grown on sugar beet pulp. A large spectrum of polysaccharide-degrading enzymes was produced by A. niger and very few ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00277-5
更新日期:2001-01-02 00:00:00
abstract::Direct esterifications using a nylon-immobilized lipase from Candida cylindracea were carried out in batch and continuous-flow reactors. The immobilized enzyme was effective in catalyzing the synthesis of ethylpropionate, isoamylpropionate, and isoamylbutyrate. With ethanol dissolved in hexane as a substrate, the maxi...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(92)90054-r
更新日期:1992-11-01 00:00:00
abstract::In the present investigation, the bacterium Escherichia coli (VM1) was isolated from a marine sediments of Ennore coastal water along southeast coast of India was exploited for its capability of Ag NPs synthesis. The derived nanoparticles were characterized employing ultraviolet-visible (UV-vis) spectroscopy, Fourier ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.09.008
更新日期:2016-12-01 00:00:00
abstract::The methyl ester of 2-benzoxazolon-3-yl-acetic acid was used as an acyl donor in the penicillin amidase-catalysed transfer reaction to 7-aminodesacetoxycephalosporanic acid. The synthesis of 7-(2-benzoxazolon-3-yl-acetamido)-desacetoxycephalosporanic acid was carried out as a kinetically controlled reaction. A charact...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(92)90144-d
更新日期:1992-06-01 00:00:00
abstract::Although the chaperon solvent plug was reported as a strategy to reduce aggregation before the column inlet in SEC (size-exclusion chromatography) protein refolding process, the appropriate position at which sample injected and the volume of the chaperon solvent plug have not been elucidated. Therefore, the detail of ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2011.05.003
更新日期:2011-07-10 00:00:00
abstract::A colorimetric platform based on enzyme-inorganic hybrid nanoflowers was used for ultraviolet-visible detection of hydrogen peroxide (H2O2). The hybrid nanoflowers were prepared via incubation at room temperature using horseradish peroxidase (HRP) and copper phosphate (Cu3(PO4)2) as the organic and inorganic component...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2020.109620
更新日期:2020-10-01 00:00:00
abstract::Operational stability under high temperature is required for enzyme application in industrial processes. Error-prone PCR and B-factor analysis were employed to enhance the thermostability of a xylanase from GH family 11 in this study. Based on the top 10 mutants screened from the random mutation libraries, mutant Xyn3...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2020.109720
更新日期:2021-02-01 00:00:00