Abstract：
:A method to quantify the impact of proteolysis on accumulation of recombinant proteins in E. coli is described. A much smaller intracellular concentration of staphylococcal protein A (SpA) (14.7 mg. g(-1)) compared to the fusion protein SpA-betagalactosidase (138 mg. g(-1)) is explained by a very high proteolysis rate constant of SpA. The SpA synthesis rate reached a maximum one hour after induction and gradually decreased to half of this value at the end of the cultivation. The decrease of the synthesis rate and the 1st order kinetics of proteolysis lead to an equilibrium between synthesis and degradation of SpA from 2 h after induction. This resulted in no further SpA accumulation in cells, though synthesis continued for at least 10 h. Similar experiments with recombinant protein ZZT2 also revealed that most of the synthesized product was degraded. The order of proteolysis kinetics depended on the concentration of the recombinant protein: at low concentrations both SpA and ZZT2 were degraded according to first order kinetics, while at high concentrations ZZT2 was degraded according to zero order kinetics. In a protease Clp mutant the degradation rate decreased and intracellular concentration of ZZT2 increased from 50 mg. g(-1) to 120 mg. g(-1). The measurements of proteolysis rate throughout the cultivation enabled calculation of a hypothetical accumulation of the product assuming complete stabilization. In this case the concentration would have increased from 50 to 280 mg. g(-1) in 11 h. Thus, this method reveals the potential to increase the productivity by eliminating proteolysis.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Rozkov A,Schweder T,Veide A,Enfors Sdoi
10.1016/s0141-0229(00)00294-5keywords:
subject
Has Abstractpub_date
2000-12-01 00:00:00pages
743-748issue
10eissn
0141-0229issn
1879-0909pii
S0141022900002945journal_volume
27pub_type
杂志文章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
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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
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abstract：:Photothermal therapy (PTT) is a minimally invasive and effective cancer treatment method and has a great potential for innovating the conventional chemotherapy approaches. Copper sulfide (CuS) exhibits photostability, low cost, and high absorption in near infrared region, and is recognized as an ideal candidate for PT...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.04.002
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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
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abstract：:Effects of pulse and step changes in major metabolite concentrations such as glucose, glutamine, lactic acid, and ammonium on behavior of T-cell hybridomas in continuous culture were investigated in order to develop a strategy to maximize specific rate of lymphokine formation. Step increases in glucose, lactic acid, a...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(90)90007-d
更新日期：1990-09-01 00:00:00
abstract：:Resveratrol is a well-known polyphenol present in red wine and exerts antioxidative and anti-carcinogenic effects on the human body. To produce resveratrol in a food-grade yeast, the 4-coumarate:coenzyme A ligase gene (4CL1) from Arabidopsis thaliana and stilbene synthase gene (STS) from Arachis hypogaea were cloned a...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2010.09.004
更新日期：2011-01-05 00:00:00
abstract：:The novel termite gut metagenomic-derived GH11 xylanase gene xyl7 was expressed in Escherichia coli BL21, and the purified XYL7 enzyme exhibited high specific activity (6340U/mg) and broad pH active range of 5.5-10.0. Directed evolution was employed to enhance the thermostability of XYL7; two mutants (XYL7-TC and XYL7...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2014.11.005
更新日期：2015-03-01 00:00:00
abstract：:The goal of this research was to develop recombinant Escherichia coli to improve fatty acid synthesis (FAS). Genes encoding acetyl-CoA carboxylase (accA, accB, accC), malonyl-CoA-[acyl-carrier-protein] transacylase (fabD), and acyl-acyl carrier protein thioesterase (EC 3.1.2.14 gene), which are all enzymes that cataly...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2011.04.001
更新日期：2011-06-10 00:00:00
abstract：:Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acid...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.01.003
更新日期：2016-04-01 00:00:00
abstract：:Three threonine aldolases (TAs) were cloned and overexpressed in Escherichia coli (Aeromonas jandaeil-allo-threonine aldolase, E. colil-threonine aldolase and Thermotoga maritimal-allo-threonine aldolase). A Design of Experiments strategy was used to identify optimal reaction conditions for each enzyme. These conditio...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2018.07.004
更新日期：2018-12-01 00:00:00
abstract：:Biosensors have been extensively studied during the last 20 years, and a myriad of laboratory biosensors have been developed. Improvements are required in biosensor design and performance before they become widely accepted in industrial process monitoring. However, as the biotechnology industry expands, biosensors may...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章,评审
doi：10.1016/0141-0229(91)90115-q
更新日期：1991-12-01 00:00:00
abstract：:The effect of treatment with various intercalating dyes on the ability to produce antibiotics in Micromonospora rosaria and Micromonospora purpurea was studied. Treatment with acriflavine resulted in a high frequency loss of antibiotic productivity in both species. In M. rosaria, the loss of antibiotic-producing abili...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(90)90128-d
更新日期：1990-08-01 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
abstract：:Recombinant plasmid pCED3 [confers beta-galactosidase production (LacZ+) and kanamycin resistance (Kmr)] in Bacillus subtilis was found to be both segregationally and structurally unstable. Since many solutions to segregational instability are already available, the problem of structural instability was specifically a...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(90)90160-r
更新日期：1990-05-01 00:00:00
abstract：:Corynebacterium glutamicum is an important industrial organism for the production of a variety of biological commodities. We discovered a promoter encoded by the gene NCgl2319 in C. glutamicum, which could be induced by benzyl alcohol, could be used as an efficient tunable expression system. In initial attempts, this ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2020.109651
更新日期：2020-11-01 00:00:00
abstract：:The effects of biological pretreatment on Pinus radiata and Eucalyptus globulus, were evaluated after exposure to two brown rot fungi Gloephylum trabeum and Laetoporeus sulphureus. Changes in chemical composition, structural modification, and susceptibility to enzymatic hydrolysis in the degraded wood were analyzed. A...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2011.08.004
更新日期：2011-10-10 00:00:00
abstract：:The family B DNA polymerase gene from the euryarchaeon Thermococcus barophilus Ch5 (Tba5) contains an open reading frame of 6198 base pairs that encodes 2065 amino acid residues. The gene is split by three inteins that must be spliced out to form the mature DNA polymerase. A Tba5 DNA polymerase gene without inteins (g...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.02.004
更新日期：2016-05-01 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：:In the paper, a green and efficient biosynthetical technique was reported for preparing cadmium sulfide (CdS) quantum dots, in which Escherichia coli (E. coli) was chosen as a biomatrix. Fluorescence emission spectra and fluorescent microscopic photographs revealed that as-produced CdS quantum dots had an optimum fluo...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.09.017
更新日期：2017-01-01 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
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更新日期：2014-12-01 00:00:00
abstract：:Human interferon alpha 2b (IFN α2b) is a type I interferon exhibiting antiviral, anti-proliferative and immunomodulatory activities. The clinical outcome of the approved recombinant human IFN α2b drugs in the market suffers from short plasma half-life, rapid clearance and other side effects. Human IFN α2b expression i...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
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更新日期：2019-09-01 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：: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
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：:Supercritical fluids offer environmental advantages over chemical solvents, while providing enhanced separation and chemical selectivity. The use of supercritical fluids for the recovery of products from biomass and the transformation of selected molecules (to add value) was studied. Free fatty acids were bio-catalyti...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2011.07.004
更新日期：2011-09-10 00:00:00
abstract：:An improved amperometric l-lactate biosensor was constructed based on covalent immobilization of lactate oxidase (LOx) from Pediococcus species onto carboxylated multiwalled carbon nanotubes (cMWCNT)/copper nanoparticles (CuNPs)/polyaniline (PANI) hybrid film electrodeposited on the surface of a pencil graphite electr...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.10.014
更新日期：2017-01-01 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：: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：:Phenylalanine ammonia-lyase (PAL) gene plays an important role in the synthesis of flavones, lignin, and other bioactive compounds in living organisms. Inonotus baumii, the only known flavone-producing filamentous fungus, is of great importance in the investigation of flavone metabolic pathways. To study the function ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2017.10.010
更新日期：2018-05-01 00:00:00
abstract：:Glutaric acid is one of the promising C5 platform compounds in the biochemical industry. It can be produced chemically, through the ring-opening of butyrolactone followed by hydrolysis. Alternatively, glutaric acid can be produced via lysine degradation pathways by microorganisms. In microorganisms, the overexpression...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2018.07.002
更新日期：2018-11-01 00:00:00
