Abstract:
:This study examines a novel method to reduce the probability of disulfide mismatches during the refolding process by the replacement of cysteines within a protein. Specifically, Cys383 of recombinant rat procarboxypeptidase B was replaced by other amino acids to increase the refolding efficiency in vitro. Mutants C383G, C383A and C383S could refold successfully, but mutants C383R, C383E, C383L and C383Y failed to refold correctly. Compared with wild type, the refolding efficiencies of mutants C383G and C383A were enhanced. The Cys383 mutations changed some of the properties of rat carboxypeptidase B. Mutants C383G, C383A had higher k(cat)/K(m) values which indicated increased catalytic abilities. And both had higher thermal stability. pH had different effects on the activities and stabilities of the mutant and wild type proteins. The studies suggested that mutating Cys383 of rat procarboxypeptidase B could improve the renaturation process by increasing the refolding efficiency. This new method could be taken as a new attempt to improve the refolding efficiency of other recombinant proteins containing disulfide bonds that are expressed as inclusion bodies. While the results also claimed that the potential effects of the substituted amino acid on the protein itself should be seriously considered in addition to its ability to reduce the probability of disulfide mismatches.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Li S,Zhang L,Wu Q,Xin A,Zhao J,Fan Ldoi
10.1016/j.enzmictec.2011.04.018subject
Has Abstractpub_date
2011-07-10 00:00:00pages
139-45issue
2eissn
0141-0229issn
1879-0909pii
S0141-0229(11)00089-5journal_volume
49pub_type
杂志文章abstract::Peptide T has a sequence (Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr) belonging to HIV envelope that is involved in the interaction with CD(4) receptor of T lymphocytes. Research of protease activities towards this peptide is very relevant for AIDS therapy. Characterization of specificity of subtilisin Carlsberg towards this ver...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(99)00170-2
更新日期:2000-03-01 00:00:00
abstract::This review focuses on the description of recently discovered esterase enzymes involved in xylan degradation (acetyl xylan, feruloyl, and p-coumaroyl esterases). The occurrence of these enzymes in various microorganisms, assays used for determination of their activity, induction and production on different substrates,...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章,评审
doi:10.1016/0141-0229(93)90078-g
更新日期:1993-06-01 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::Trehalose diesters are Gemini-type surfactants that might have better surface activity than conventional surfactants. A one-step method for the preparation of trehalose unsaturated fatty acid diesters has been successfully developed. The yield of trehalose diester of different unsaturated fatty acids was between 78 % ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2020.109516
更新日期:2020-05-01 00:00:00
abstract::Glucose is the typical carbon source for producing microbial polyunsaturated fatty acids (PUFA) with single cell microorganisms such as thraustochytrids. We assessed the use of a fish oil derived glycerol by-product (raw glycerol), produced by a fish oil processing plant, as a carbon source to produce single cell oil ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2010.11.008
更新日期:2011-03-07 00:00:00
abstract::Catalase was covalently immobilized onto florisil via glutaraldehyde (GA) and glutaraldehyde+6-amino hexanoic acid (6-AHA) (as a spacer arm). Immobilizations of catalase onto modified supports were optimized to improve the efficiency of the overall immobilization procedures. The V(max) values of catalase immobilized v...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2011.09.002
更新日期:2011-12-10 00:00:00
abstract::Filamentous fungal secretomes comprise highly dynamic sets of proteins, including multiple carbohydrate active enzymes (CAZymes) which are able to hydrolyze plant biomass polysaccharides into products of biotechnological interest such as fermentable sugars. In recent years, proteomics has been used to identify and qua...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章,评审
doi:10.1016/j.enzmictec.2017.08.007
更新日期:2018-02-01 00:00:00
abstract::l-asparaginase is a pharmaceutically and industrially important enzyme as it has potential to treat different cancers and inhibit acrylamide formation in fried and baked food products. In the present study, an attempt was made to screen for new and novel l-asparaginase producers using a widely applied phenol red and b...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.08.004
更新日期:2017-12-01 00:00:00
abstract::This work presents the purification and characterization of an extracellular alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) produced by a new lactic acid bacterium: Lactobacillus manihotivorans able to produce L(+) lactic acid from starch. The molecular weight was found to be 135 kDa. The temperature ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00230-1
更新日期:2000-09-01 00:00:00
abstract::The cellulase and xylanase genes of filamentous Trichoderma fungi exist under carbon catabolite repression mediated by the regulator carbon catabolite repressor (CREI). Our objective was to find the role of CREI in a cellulase-hyperproducing mutant of Trichoderma koningii, and address whether enzyme production can be ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2013.04.007
更新日期:2013-07-10 00:00:00
abstract::A novel kinetic method was developed for the quantitation of α-ketoglutaric acid (AKG) in cardioplegic solution and athletic supplements. The assay relied on an enzymatic transamination of AKG and d-4-hydroxyphenylglycine to form 4-hydroxybenzoylformic acid and l-glutamic acid using d-phenylglycine aminotransferase. S...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2018.05.013
更新日期:2018-09-01 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::The activities (at pH 7 and 50 degrees C) of purified EGV (Humicola insolens) and CenA (Cellulomonas fimi) were determined on cotton fabrics at high and low levels of mechanical agitation. Similar activity measurements were also made by using the core domains of these cellulases. Activity experiments suggested that th...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00205-2
更新日期:2000-08-01 00:00:00
abstract::The D-amino acid oxidase cDNA gene (daao) of Trigonopsis variabilis was prepared by reverse transcriptase-polymerase chain reaction (PCR) and cloned into Escherichia coli expression vector, pTrc99A, under the control of tac promoter. Expression of daao gene significantly affected the growth and morphology of E. coli. ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00247-7
更新日期:2000-10-01 00:00:00
abstract::In the present work, NAD(+)-dependent formate dehydrogenase (FDH), encoded by fdh gene from Candida boidinii was successfully displayed on Escherichia coli cell surface using ice nucleation protein (INP) from Pseudomonas borealis DL7 as an anchoring protein. Localization of matlose binding protein (MBP)-INP-FDH fusion...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.06.002
更新日期:2016-09-01 00:00:00
abstract::Continuous culture and fed-batch fermentations were used to test the behavior of the system Bacillus subtilis DN1885(pCH7) that synthesizes a recombinant beta-1,4-endoglucanase. Continuous culture experiments were focused on the study of the instability aspects of the system as well as determination of the biomass gro...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(00)00203-9
更新日期:2000-08-01 00:00:00
abstract::A method is described for construction of a highly sensitive electrochemical biosensor for detection of glutamate. The biosensor is based on covalent immobilization of glutamate oxidase (GluOx) onto polypyrrole nanoparticles and polyaniline composite film (PPyNPs/PANI) electrodeposited onto Au electrode. The enzyme el...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.02.001
更新日期:2014-04-10 00:00:00
abstract::This work describes for the first time the green synthesis of neopentyl glycol diheptanoate in a solvent-free medium via an enzymatic pathway. The process has been carried out in an open-air reactor in order to ease water removal through evaporation and shift the chemical equilibrium towards product formation. The inh...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2019.109400
更新日期:2020-01-01 00:00:00
abstract::Even though many enzyme processes occur at the interface of an insoluble substrate, these reactions are generally much less studied than homogenous enzyme reactions in the aqueous bulk. Interfacial (or heterogeneous) enzyme reactions involve several reaction steps, and the established experimental approach to elucidat...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.06.009
更新日期:2017-10-01 00:00:00
abstract::Zygosaccharomyces rouxii is an important yeast in the formation of flavor in soy sauce. In this study, we investigated the separate effects of exogenous threonine, cystathionine, and the branched-chain amino acids on the metabolism of Z. rouxii. The addition of these amino acids had significant effects on both Z. roux...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(99)00165-9
更新日期:2000-02-01 00:00:00
abstract::The fatty acid methyl esters and fatty acid ethyl esters are known as biodiesels which are considered to be renewable, nontoxic and biodegradable biofuels. However, the conventional biodiesels show a high crystallization temperature which is one of the most critical obstacles against the widespread biodiesel usage. Th...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.03.012
更新日期:2017-07-01 00:00:00
abstract::The mechanical properties of TB/C3 hybridoma cells taken from a continuous culture were measured by micromanipulation. The culture conditions were constant except for the presence or absence of Pluronic F-68 in the medium. It was found that the mean bursting membrane tension and the mean elastic area compressibility m...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(92)90081-x
更新日期:1992-12-01 00:00:00
abstract::Phytate is the major storage form of phosphorus in plants. It is present in cereals and raw materials of vegetable origin used in animal and human diets. However, non-ruminant animals have little phytase activity in their guts and, therefore, cannot digest phytate. As a result, almost all dietary phytate is discharged...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2020.109533
更新日期:2020-06-01 00:00:00
abstract::This study aimed to develop viable enzymes for bioconversion of resveratrol-glucoside into resveratrol. Out of 13 bacterial strains tested, Lactobacillus kimchi JB301 could completely convert polydatin into resveratrol. The purified enzyme had an optimum temperature of 30-40°C and optimum pH of pH 5.0 against polydati...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.09.001
更新日期:2014-12-01 00:00:00
abstract::A reactive water-soluble polymer was synthesized by copolymerizing N-isopropylacrylamide and glycidyl acrylate. The reactive polymer could react with the amino groups of enzymes/proteins or other ligands to form an affinity polymer. As a model, the reactive polymer was allowed to react with paraaminobenzamidine, a str...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(90)90005-b
更新日期:1990-09-01 00:00:00
abstract::This study aimed to improve the thermostability of a newly cloned tyrosinase from Streptomyces kathirae SC-1. The POPMuSiC algorithm was applied to predict the folding free energy change (ΔDG) of amino acid substitution. Site-directed mutagenesis was used to construct mutants (Q7K, G234P, and Q7K/G234P), and the mutan...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.06.002
更新日期:2015-09-01 00:00:00
abstract::Cystathionine γ-lyase (CGL) is a key enzyme in the methionine-cysteine cycle in all living organisms forming cysteine, α-ketobutyrate and ammonia via homocysteine and cystathionine intermediates. Although, human and plant CGLs have been extensively studied at the molecular and mechanistic levels, there has been little...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.08.004
更新日期:2015-12-01 00:00:00
abstract::The selected thermotolerant, lactose-utilizing yeast strain Kluyveromyces marxianus NBIMCC 8362 possesses high specific d-amino acid oxidase activity (60Ug(-1)), which was increased nine-fold (545Ug(-1)) by design of the growth medium and conditions for d-amino oxidase induction. Applying an optimized simple and rapid...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2007.08.010
更新日期:2008-01-01 00:00:00
abstract::Data and a semi-empirical model are presented that describe the affinity interaction of yeast cells with a Concanavalin A derivatised surface. The model uses 3 parameters to describe the time course of cell attachment from a flowing suspension of yeast cells, over a range of flow rates, and gives an effective global f...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/s0141-0229(99)00161-1
更新日期:2000-02-01 00:00:00
abstract::Advantages in maltose fermentation at extremely low temperatures (5-10°C) using an alcohol resistant and cryotolerant yeast strain (Saccharomyces cerevisiae AXAZ-1) immobilized on porous cellulosic material (or tubular cellulose, TC), produced by delignification of wood sawdust, are reported. Pure maltose and glucose ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.08.007
更新日期:2014-11-01 00:00:00