Abstract：
:In the current study, a three-tiered mutagenesis strategy was employed to simultaneously improve the thermostability and activity of halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) by engineering the last ten amino acids (Met245∼Glu254) of its C-terminal region. Initially, truncated mutagenesis results displayed that C-terminal deletions decreased the thermostability and/or activity of HheC. Then ten residues were subjected to single-site saturation mutagenesis, resulting in 20 beneficial single-point variants related to the thermostability or activity of HheC. The results clearly indicated that residues Met252∼Glu254 and Trp249 are crucial for regulating enzyme thermostability and activity, respectively. Finally, the beneficial substitutions were combined using efficient multi-site combinatorial mutagenesis approaches, leading to an outstanding variant PX14 (Trp249Pro/Met252Leu/Pro253Asp), which had a 17.8-fold higher half-life and a 4.0-fold higher kcat value than that of wild-type HheC. These results indicated that the C-terminal residues play an important role in modulating both the thermostability and activity of HheC.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Wang X,Han S,Yang Z,Tang Ldoi
10.1016/j.jbiotec.2015.08.013subject
Has Abstractpub_date
2015-10-20 00:00:00pages
92-8eissn
0168-1656issn
1873-4863pii
S0168-1656(15)30094-8journal_volume
212pub_type
杂志文章abstract：:Saccharum spontaneum L., a wild relative of sugarcane, is known for its adaptability to environmental stresses, particularly cold stress. In the present study, an attempt was made for transcriptome profiling of the low temperature (10°C) tolerant S. spontaneum clone IND 00-1037 collected from high altitude regions of ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.05.036
更新日期：2016-08-10 00:00:00
abstract：:Protein engineering has been employed to investigate the effect of specific amino acid changes on the targeting of heterologous proteins to the outer cell surface of the Gram-positive bacterium Staphylococcus xylosus. Three different variants, corresponding to a 101 amino acid region of the major glycoprotein (G prote...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(95)00081-z
更新日期：1995-10-16 00:00:00
abstract：:Biosurfactants are surface-active agents produced by microorganisms and show increasing significance in various industrial applications. A great variety of these secondary metabolites are described to occur within actinomycetes, amongst trehalose lipids and oligosaccharide lipids produced by the family Tsukamurellacea...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.06.424
更新日期：2015-10-10 00:00:00
abstract：:We cloned and characterized a preprochymosin cDNA from the abomasum of milk-fed kid goats. This cDNA contained an open reading frame that predicts a polypeptide of 381 amino acid residues, with a signal peptide and a proenzyme region of 16 and 42 amino acids, respectively. Comparison of the caprine preprochymosin sequ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.06.002
更新日期：2004-10-19 00:00:00
abstract：:Fiber morphology and crystalline structure of poly[(R)-3-hydroxybutyrate] (P(3HB)) and stereocomplexed poly(lactide) (PLA) nanofibers were investigated by using scanning and transmission electron microscopies and X-ray and electron diffractions. In the P(3HB) nanofibers spun from less than 1 wt% 1,1,1,3,3,3-hexafluoro...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.03.019
更新日期：2007-11-01 00:00:00
abstract：:Microarray analysis has become a popular and routine method in functional genomics. It is typical for such experiments to involve a small number of replicates, which causes unreliable estimates of the sample variance. Microarrays have fostered the development of new statistical methods to analyze data resulting from e...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.01.009
更新日期：2009-03-10 00:00:00
abstract：:A gene encoding mature human insulin-like growth factor II (IGF-II) was constructed from the modified IGF-II cDNA sequence and two double-stranded synthetic oligodeoxynucleotide linkers. It was fused to a truncated lacZ gene such that IGF-II was expressed as part of C-terminus of beta-galactosidase. This fused lacZ'-I...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(90)90077-o
更新日期：1990-03-01 00:00:00
abstract：:To explore the utility of glycosyltransferases as novel biocatalysts, we isolated the glycosyltransferase genes CaUGT2 and SbUGTA1 from Catharanthus roseus and Starmerella bombicola, respectively and heterologously expressed them in Escherichia coli. The purified recombinant proteins were assayed with a variety of sma...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.10.011
更新日期：2015-12-20 00:00:00
abstract：:Kinetic models have a great potential for metabolic engineering applications. They can be used for testing which genetic and regulatory modifications can increase the production of metabolites of interest, while simultaneously monitoring other key functions of the host organism. This work presents a methodology for in...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.01.005
更新日期：2016-03-20 00:00:00
abstract：:The extraction of 13-hydroperoxide-lyase activity from mint leaves as well as its use for C6-aldehyde production was studied in this work. The enzyme cleaves 13(S)-hydroperoxy-C18 fatty acids into C6-aldehyde and C12-oxo-acid. Two mint species were tested: Mentha veridis and Mentha pulegium. The headspace injection me...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.03.005
更新日期：2004-07-01 00:00:00
abstract：:ACC deaminase producing Plant growth promoting rhizobacteria (PGPR) offers a great promise for ameliorating the negative impacts of salinity stress manifested on plants. In this context, 28 rhizospheric bacteria associated with ACC deaminase potential (198-1069 nmol α-ketobutyrate mg protein-1 h-1) were isolated from ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2020.10.024
更新日期：2020-12-20 00:00:00
abstract：:The circadian clock schedules processes in microalgae cells at suitable times in the day/night cycle. To gain knowledge about these biological time schedules, Neochloris oleoabundans was grown under constant light conditions and nitrogen limitation. Under these constant conditions, the only variable was the circadian ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.07.016
更新日期：2014-10-10 00:00:00
abstract：:The effects of a deletion of the amtR gene, encoding the master regulator of nitrogen control in Corynebacterium glutamicum, were investigated by metabolome and transcriptome analyses. Compared to the wild type, different metabolite patterns were observed in respect to glycolysis, pentose phosphate pathway, citric aci...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.10.009
更新日期：2009-03-10 00:00:00
abstract：:Zeolitic imidazole frameworks (ZIFs) with tunable pore sizes and high surface areas have recently used as an effective support for immobilizing enzymes. However, the instability in the aqueous acidic environment has limited their practical applications in some cases. In this work, we develop a novel catalase/ZIFs comp...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.09.010
更新日期：2019-12-20 00:00:00
abstract：:In this work, the ethanolysis of triglycerides catalyzed by immobilized lipase was studied, focusing on the secondary reaction of acyl migration. The catalytic tests were performed in a solvent-free reaction medium using Novozym 435 as biocatalyst. The selected experimental variables were biocatalyst loading (5-20mg),...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.01.001
更新日期：2016-02-20 00:00:00
abstract：:Laccase from the white rot fungus Coriolopsis polyzona was immobilized for the first time through the formation of cross-linked enzyme aggregates (CLEAs). Laccase CLEAs were produced by using 1000g of polyethylene glycol per liter of enzyme solution as precipitant and 200muM of glutaraldehyde as a cross-linking agent....
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.07.948
更新日期：2007-10-15 00:00:00
abstract：:Cannabinoids are secondary natural products from the plant Cannabis sativaL. Therapeutic indications of cannabinoids currently comprise a significant area of medicinal research. We have expressed the Δ9-tetrahydrocannabinolic acid synthase (THCAS) and cannabidiolic acid synthase (CBDAS) recombinantly in Komagataella p...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.07.031
更新日期：2018-10-20 00:00:00
abstract：:Recombinant Escherichia coli clones encoding between 0 and 6 copies of the mammalian cytochrome b5 gene were subjected to pyrolysis mass spectrometry (PyMS). To deconvolute the pyrolysis mass spectra so as to obtain quantitative information on the amount of cytochrome b5 produced fully-interconnected feedforward artif...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(94)90088-4
更新日期：1994-05-15 00:00:00
abstract：:A Gram-negative, rod-shaped, non-motile, gamma and UV radiation resistant bacterium Hymenobacter radioresistens DG25B was isolated from a soil sample collected in South Korea. The complete genome sequence of H. radioresistens DG25B consists of one circular chromosome (3,874,646 bp). The bacterium was isolated from gam...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.11.015
更新日期：2016-01-10 00:00:00
abstract：:beta-galactosidase reporter plasmids containing different viral or minimal promoters are commonly used to correct variable transfection efficiencies in transient transfection experiments. The transcriptional activity of these promoters is thought to be stable under most circumstances. To determine if expression of bet...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(01)00277-2
更新日期：2001-07-12 00:00:00
abstract：:Recombinant human granulocyte colony stimulating factor (rhG-CSF) and its PEGylated product "mono-PEG20-GCSF" have already been widely used for treatment of all kinds of neutropenia. However, the high required dosage of mono-PEG20-GCSF made it relatively expensive in clinical use. We postulated that an N-terminal site...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.05.012
更新日期：2009-07-15 00:00:00
abstract：:In this report, we describe the results of a rat pituitary cell culture experiment done on STS-65 in which the effect of cell feeding on the release of the six anterior pituitary hormones was studied. We found complex microgravity-related interactions between the frequency of cell feeding and the quantity and quality ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(96)01373-9
更新日期：1996-06-27 00:00:00
abstract：:Selected phage clones expressing a peptide with high binding affinity for recombinant human lactoferrin or von Willebrand factor (vWF) were covalently coupled to macroporous poly(dimethylacrylamide) monolithic column. Large pore size (10-100 microm) of macroporous poly(dimethylacrylamide) makes it possible to couple l...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.06.021
更新日期：2007-09-15 00:00:00
abstract：:A novel method that relies on the decoupling of the energy production and biosynthesis processes was used to characterise the maintenance, cell lysis and growth processes of Nitrosomonas sp. A Nitrosomonas culture was enriched in a sequencing batch reactor (SBR) with ammonium as the sole energy source. Fluorescent in ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.04.017
更新日期：2006-11-10 00:00:00
abstract：:The cyclic amino acid ectoine is a compatible solute serving as a protective substance against osmotic stress. Ectoine finds various applications due to its moisturizing effect. To avoid the disadvantages of the prevailing so-called "bacterial milking ectoine production process" caused by the high salt concentration, ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.04.039
更新日期：2017-09-20 00:00:00
abstract：:There are many examples of enzymes that share substrates or cofactors in a cyclic manner. Techniques have been developed that use cyclic enzyme systems to assay quantitatively small amounts of biochemical substances (cofactor, substrate), however, only a few studies of the control of these systems have been published....
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/0168-1656(92)90064-g
更新日期：1992-06-01 00:00:00
abstract：:A PHA producing microbial culture dominated by Plasticicumulans acidivorans was enriched in a pilot plant using fermented wastewater from the Mars candy bar factory. The pilot plant comprised (1) anaerobic fermentation, (2) enrichment of a PHA-producing microbial community and (3) accumulation for maximization of the ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.10.022
更新日期：2014-12-20 00:00:00
abstract：:Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in ext...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.10.015
更新日期：2010-01-15 00:00:00
abstract：:Dual nutrient limited growth, the control of the cell growth rate (kinetic aspect) or the restriction of the amount of biomass (stoichiometric aspect) by two nutrients at the same time, is a relatively unknown ability of the microorganisms and consequently, still not mentioned in textbooks to date. Nevertheless, multi...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/j.jbiotec.2004.03.030
更新日期：2004-09-30 00:00:00
abstract：:The importance of RNA-based regulation is becoming more and more evident. Genome-wide sequencing efforts have shown that the majority of the DNA in eukaryotic genomes is transcribed. Advanced high-throughput techniques like CLIP for the genome-wide detection of RNA-protein interactions have shown that post-transcripti...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/j.jbiotec.2017.05.019
更新日期：2017-11-10 00:00:00
