Abstract：
:We studied, for the first time, characterization of the invertase expressed in the methylotrophic yeasts Hansenula polymorpha and Pichia pastoris in terms of enzyme conformational stability and structural behaviour induced by temperature as a function of pH using enzymic assays, differential scanning calorimetry, fluorescence and CD. The enzyme produced in both hosts was very stable over a broad range of pH values, keeping its enzymic activity and structure above 60 degrees C. Thermal denaturation, as measured by differential scanning calorimetry, was always irreversible. However, the fact that scanning rate had no effect on the calorimetric curves gave us the chance to analyse the data from a thermodynamic point of view. The conformational stabilities were essentially identical under the experimental conditions studied, but stability was always slightly higher in the enzyme expressed in H. polymorpha. This fact indicates that the greater degree of glycosylation of this enzyme form contributed to its increased global stability. Reactivation upon heating at 80 degrees C depends on protein concentration, suggesting that irreversibility could be associated with slow refolding kinetics at high protein concentration.
journal_name
Biotechnol Appl Biochemjournal_title
Biotechnology and applied biochemistryauthors
Acosta N,Beldarraín A,Rodríguez L,Alonso Ydoi
10.1042/ba20000064subject
Has Abstractpub_date
2000-12-01 00:00:00pages
179-87issue
3eissn
0885-4513issn
1470-8744journal_volume
32pub_type
杂志文章abstract：:Two monoclonal antibodies against human myeloperoxidase, designated 3-2H3 and 4-2C11, were produced and characterized. Both bound to the native enzyme, but neither bound to the denatured enzyme or to its two denatured subunits. 4-2C11 bound to the three types of leukocyte myeloperoxidase, I, II, and III, as well as to...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1986-12-01 00:00:00
abstract：:Some epidemiological studies have suggested possible associations between exposure to extremely low-frequency electromagnetic fields (ELF-EMFs) and various diseases. Recently, ELF-EMF has been considered as a therapeutic agent. To support ELF-EMF use in regenerative medicine, in particular in the treatment of skin inj...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1495
更新日期：2017-05-01 00:00:00
abstract：:Acetone-butanol-ethanol (ABE) fermentation was performed with sugarcane bagasse (SCB) hydrolysate using Clostridium beijerinckii strains. A cost-effective SCB medium was developed with no enzymatic hydrolysis and no supplementation of extra carbon source or expensive nitrogen source. One of the C. beijerinckii strains...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1865
更新日期：2020-09-01 00:00:00
abstract：:The aim of the present study was to determine both the minimal and the optimal conditions under which heat treatment is effective for enhancing 3D (three-dimensional)-like cell proliferation. C3H10T1/2 mouse fibroblasts were cultured with hydroxyapatite granules for 10 weeks after heat treatment at 40, 41.5, 43, 44 or...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20060200
更新日期：2007-05-01 00:00:00
abstract：:A new system equipped with a computer-controlled multiple activity analyzer has been developed for the efficient purification of multiple enzymes. The system consists of the following units: conventional enzyme fractionation system with a peristaltic pump, liquid chromatographic column, fraction collector, and uv moni...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1986-12-01 00:00:00
abstract：:Recently, the development of "green" methods for fabrication of silver nanoparticles (Ag-NPs) has been emphasized, in view of their environmental safety, feasibility, and low cost. In this study, a serine protease, EuP-82 from Euphorbia cf. lactea latex, was used to fabricate silver chloride nanoparticles (AgCl-NPs) i...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1638
更新日期：2018-07-01 00:00:00
abstract：:A pathway for the synthesis of the repeating tetrasaccharide units in gellan gum from Pseudomonas elodea is proposed. The enzymes presumed to be involved in the synthesis of the activated precursors UDP-glucose, TDP-rhamnose, and UDP-glucuronic acid were detected and assayed in crude cell extracts of the gellan-produc...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1991-12-01 00:00:00
abstract：:We have analysed the status and transcriptional activity of the bovine papillomavirus-I (complete BPV-I genome)-based expression vector pCES in CI27i-cell-line-derived 3TI cells used for the industrial production of recombinant human erythropoietin (rhuEpo). Complete tandem head-to-tail integration of about 600 vector...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1996-02-01 00:00:00
abstract：:The use of the recombinant BoNT/A-Hc (carboxylic domain of the Clostridium botulinum neurotoxin heavy chain) has been proposed as a vaccine candidate for botulism. This fragment contains the principle protective antigenic determinant. In the present study, in order to maximize recombinant protein expression, after ver...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20090315
更新日期：2010-05-24 00:00:00
abstract：:As a highly contagious and potentially fatal disease of dogs, canine parvovirus type 2 (CPV-2) usually causes severe myocarditis and gastroenteritis, while vaccine injection has greatly reduced the incidence of CPV-2 diseases. However, there is currently a lack of simple and effective method for quantitative detection...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1967
更新日期：2020-06-13 00:00:00
abstract：:This review article focuses on the use of the methylotrophic yeast Pichia pastoris as a recombinant protein-expression system. P. pastoris is a useful system for the expression of milligram-to-gram quantities of a protein, which can be scaled up to fermentation to meet greater demands. Compared with mammalian cells, P...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章,评审
doi：
更新日期：1999-10-01 00:00:00
abstract：:A decrease in the specific activity of an enzyme is commonly observed when the enzyme is inappropriately handled or is stored over an extended period. Here, we reported a functional transition of an FMN-bound diaphorase (FMN-DI) that happened during the long-term storage process. It was found that FMN-DI did not simpl...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1607
更新日期：2018-05-01 00:00:00
abstract：:An inducible esterase has been isolated from a liquid culture of Aspergillus niger grown on sugar-beet pulp. The enzyme was active on methyl esters of cinnamic acids, caffeic > p-coumaric > ferulic, and is therefore termed a cinnamoyl esterase. The enzyme was not active on methyl sinapinate, a good substrate for ferul...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1996-06-01 00:00:00
abstract：:Cordyceps sinensis (Chinese caterpillar fungus) is a fungus parasitic on the larvae of Lepidoptera and has been considered to be a precious tonic food and herbal medicine since ancient times in China. Recently, some fungal strains have been isolated from the fruiting bodies of wild C. sinensis, and some of them have b...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20040183
更新日期：2005-08-01 00:00:00
abstract：:Synthetic membranes containing 10% acrylamide units were subjected to activation with formaldehyde at pH 7.5 and 45 degrees C. Trypsin, invertase, and urease were bound to this activated membrane and the kinetic properties of immobilized enzymes were studied. The permeability of the membrane for distilled water manife...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1991-02-01 00:00:00
abstract：:Catalytic properties of Escherichia coli polyphosphate kinase (EC 2.7.4.1), a promising enzyme for use in ATP regeneration (Hoffman, et al., 1988, Biotechnol. Appl. Biochem. 10, 107-117), are reported here. E. coli polyphosphate kinase (PPK) is broadly active in the pH range 5.5 to 8.5, having an optimal Vmax at pH 7....
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1992-04-01 00:00:00
abstract：:Medulloblastoma is the most common malignant pediatric brain tumor in children. GPR137 is a ubiquitously expressed gene in the central nervous system. It has been reported that GPR137 modulates malignant proliferation of glioma cells. However, the relationship between GPR137 and medulloblastoma is still unknown. In th...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1331
更新日期：2015-11-01 00:00:00
abstract：:This paper is concerned with the optimization of submerged culture conditions for mycelial growth and exo-biopolymer production by Auricularia polytricha by one-factor-at-a-time and uniform design (UD) methods. First, the one-factor-at-a-time method was adopted to investigate the effects of environmental factors (i.e....
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20030020
更新日期：2003-10-01 00:00:00
abstract：:Gene doping can be easily concealed since its product is similar to endogenous protein, making its effective detection very challenging. In this study, we selected insulin-like growth factor I (IGF-I) exogenous gene for gene doping detection. First, the synthetic IGF-I gene was subcloned to recombinant adeno-associate...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1518
更新日期：2017-07-01 00:00:00
abstract：:A simplified and low-cost method, boiling lysis, was developed and used to purify the truncated DNA polymerase Stoffel fragment after overproduction of the enzyme in Escherichia coli. The method is based on the thermostable properties of the Stoffel fragment. Conditions such as the boiling time and number of cycles em...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20070114
更新日期：2008-06-01 00:00:00
abstract：:The plant polymer lignin is the greatest source of aromatic chemical structures on earth. Hence, the chemically diverse lignin monomers are valuable raw materials for fine chemicals, materials synthesis, and food and flavor industries. However, extensive use of this natural resource is hampered by the large number of ...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1222
更新日期：2014-11-01 00:00:00
abstract：:Renewable energy resources are considered to be promising for the development of a sustainable circular economy. Among various alternatives, the microbial route for various biofuels production is quite lucrative. Use of cellulose and lignocellulose for methane, H2 , organic acids, ethanol, and cellulase has been explo...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章,评审
doi：10.1002/bab.1720
更新日期：2019-03-01 00:00:00
abstract：:A deletion htpR mutant of Escherichia coli has been constructed on the basis of site-directed mutagenesis. To this end, the chromosomal allele of htpR gene was substituted by a mutant allele introduced into the cell with a recombinant plasmid. The htpR mutant is characterized by a reduced level of proteolysis and ther...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1988-10-01 00:00:00
abstract：:The hybridization of the native and chemically inactivated aspartase from Escherichia coli was studied. Preparations of the tetrameric enzyme obtained by mixing the native and N-ethylmaleimide (NEM)-inactivated aspartase in 4 M guanidine-HCl followed by 51-fold dilution at room temperature retained catalytic activity....
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1990-04-01 00:00:00
abstract：:A simple, specific, sensitive and reproducible ELISA has been developed to quantify the level of CPS (capsular polysaccharide) production in supernatants of Streptococcus pneumoniae cell cultures. CPSs from Strep. pneumoniae have been widely used as vaccine antigens. The quantification method is based on two type-23F ...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20060007
更新日期：2006-05-01 00:00:00
abstract：:The hydrolysis kinetics of native and denatured haemoglobin, using pepsin immobilized on aluminium oxide, was studied in order to produce hydrolysates containing bioactive peptides. Pepsin was immobilized on acidic alumina and on 2-ethanolamine- O -phosphate (2-EAOP)-modified acidic alumina. Surface charge of the two ...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20030131
更新日期：2004-04-01 00:00:00
abstract：:A microbial fuel cell is a sustainable and environmental-friendly device that combines electricity generation and wastewater treatment through metabolic activities of microorganisms. However, low power output from inadequate electron transfer to the anode electrode hampers its practical implementation. Nanocomposites ...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1928
更新日期：2020-04-21 00:00:00
abstract：:Penicillium ochrochloron Biourge, which was isolated from rotten sweet potato, can produce plant cell wall-degrading enzymes (PCWDEs) with high viscosity reducing capability for ethanol production using fresh sweet potato tubers as feedstock. The enzyme preparation was characterized by a broad enzyme spectrum includin...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1190
更新日期：2014-07-01 00:00:00
abstract：:A simple and cost-effective method is described to sequence double-stranded DNA (< 100 ng) using a very small amount of fluorescent sequencing mix (one-tenth of manufacturer-recommended amount). The modification has significant advantages over conventional protocols and can be used to sequence, using a single sequenci...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1997-08-01 00:00:00
abstract：:A novel iron superoxide dismutase (Fe-SOD) gene from Jatropha curcas was cloned and expressed in Escherichia coli BL21 (DE3). This recombinant enzyme was isolated by a combined procedure involving immobilized metal-ion affinity chromatography and ion-exchange chromatography. The apparent molecular mass of this purifie...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1030
更新日期：2012-09-01 00:00:00
