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 mutant, and wild-type enzymes were expressed in Escherichia coli (DE3). Compared to the wild-type tyrosinase, all three mutant enzymes showed improved thermal properties. The mutant with combined substitution (Q7K/G234P) showed the most pronounced shifts in temperature optima, about 10 °C upward, and the half-life for thermal inactivation at 60 °C, and melting temperatures were increased by 3 times and approximately 10 °C, respectively. Finally, the mechanisms responsible for the increased thermostability were analyzed through comparative analysis of structure models. The structure-based rational design strategies in this study may also provide further insight into the thermostability of other industrial enzymes and suggest further potential industrial applications.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Guo J,Rao Z,Yang T,Man Z,Xu M,Zhang X,Yang STdoi
10.1016/j.enzmictec.2015.06.002subject
Has Abstractpub_date
2015-09-01 00:00:00pages
54-60eissn
0141-0229issn
1879-0909pii
S0141-0229(15)30010-7journal_volume
77pub_type
杂志文章abstract：:Phenethyl ester is an important contributors to food, drinks, cosmetics and perfumery. Phenethyl acetate was synthesized enzymatically from phenethyl alcohol with acyl donors catalyzed by Novozym 435 from Candida antarctica lipase B. Through acyl donor screening, acetic anhydride and vinyl acetate were found to be the...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2017.02.004
更新日期：2017-05-01 00:00:00
abstract：:We report a new colorimetric assay to quantify endo-polygalacturonase activity, which hydrolyzes polygalacturonic acid to produce smaller chains of galacturonate. Some of the reported polygalacturonase assays measure the activity by detecting the appearance of reducing ends such as the Somogyi-Nelson method. As a resu...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2010.04.004
更新日期：2011-02-08 00:00:00
abstract：:We demonstrate the feasibility of growing crystals of protein in volumes as small as 1 nanoliter. Advances in the handling of very small volumes (i.e. through inkjet and other technologies) open the way towards fully automated systems. The rationale for these experiments is the desire to develop a system that speeds u...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(01)00496-3
更新日期：2002-03-13 00:00:00
abstract：:Crocin, one of the major carotenoid pigments of Crocus sativus (saffron), is responsible for antioxidant activity, neuroprotection, and the inhibition of tumor cell proliferation. In order to improve the functionality of crocin, α-glucosyl-(1→6)-trans-crocins (C-Gs) were synthesized using sucrose and dextransucrase fr...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2020.109630
更新日期：2020-10-01 00:00:00
abstract：:3-Mercaptopyruvate (3MPy), a structural analog of 3-mercaptopropionic acid, is a precursor compound for biosynthesis of polythioesters in bacteria. The cost-effectiveness and sustainability of the whole process could be greatly improved by using the cysteine degradation pathway for an intracellular supply of 3MPy. Tra...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2017.01.004
更新日期：2017-04-01 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：: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：:Based on a differential pre-screening of 44 white-rot fungi on a lignocellulose-supplemented minimal medium, four basidiomycetes were selected for further study: Ganoderma lucidum, Polyporus brumalis, Polyporus ciliatus and Trametes versicolor. Only G. lucidum was able to grow vividly on malt extract or minimal media ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2013.08.003
更新日期：2013-12-10 00:00:00
abstract：:In this study, we report the enzymatic production of glycerol acetate from glycerol and methyl acetate. Lipases are essential for the catalysis of this reaction. To find the optimum conditions for glycerol acetate production, sequential experiments were designed. Type of lipase, lipase concentration, molar ratio of re...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2014.11.004
更新日期：2015-02-01 00:00:00
abstract：:A new bi-modular, wide pH spectrum and highly active xylanase KRICT PX3 (JF320814) isolated from Paenibacillus terrae HPL-003 (KCTC11987BP) has been cloned and expressed in Escherichia coli. Purified recombinant xylanase KRICT PX-3 (1,620 bp, 540aa, NCBI accession number JF320814) showed highly active at 55°C in pH 4....
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2013.09.002
更新日期：2014-01-10 00:00:00
abstract：:A novel automated image analysis method is described for characterizing the viability and morphology of animal cells from suspension cultures. With the aid of the exclusion dye Trypan Blue, the total and viable cell counts and the percentage of dead cells present are found. The area, perimeter, equivalent diameter, an...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(94)90106-6
更新日期：1994-01-01 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：: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：:Cytochrome P450 enzyme preparations were obtained from phenobarbital-treated rats. On immobilization in Romicon PM 10 polysulfone anisotropic hollow fibers, 20-25% of the original activity remained. The immobilization procedure did not significantly alter the Km of the preparations when 7-ethoxycoumarin was the substr...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(93)90031-v
更新日期：1993-02-01 00:00:00
abstract：:The primary aim of this study was to decipher the catalytic functions of the NHase with wide substrate spectra from Rhodococcus ruber CGMCC3090 by computer modeling and substrate docking. 3D structure model of the enzyme was built by computer modeling to obtain the optimal structure. The larger binding site cavity (55...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2012.11.004
更新日期：2013-02-05 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：: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 cell wall integrity and stress response component (WSC) domain was first described in the Wsc-family protein of the yeast Saccharomyces cerevisiae, and later found in diverse eukaryotic organisms. Due solely to their presence in the Wsc-family proteins working as a plasma membrane sensor for surface stress and in ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2019.02.009
更新日期：2019-06-01 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：:We generated a bifunctional enzyme chimera containing the xylanase and lichenase coupled with SpyTag between them. Meanwhile, we generated another chimera containing SpyCatcher and elastin-like polypeptides (ELPs). As ELPs could bond to the xylanase-lichenase chimera through SpyTag/SpyCatcher spontaneous reaction in m...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2018.04.007
更新日期：2018-08-01 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：:Spodoptera frugiperda insect cells (IPLB-Sf21-AE) (Sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. In the present study the culture conditions of these insect cells were studied to establish high-den...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(94)90022-1
更新日期：1994-06-01 00:00:00
abstract：:Growth conditions for mass production of luciferase-bacterial magnetic particles (BMPs) by a recombinant Magnetospirillum magneticum AMB-1 were investigated in a pH-regulated fed-batch culture system. Enrichment of growth medium with L-cysteine, yeast extract and polypeptone enhanced both bacterial growth and BMP prod...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(01)00343-x
更新日期：2001-07-05 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：: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：: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：:The effect of hexoses (glucose and galactose) addition to the feed xylose mineral medium of Debaryomyces hansenii chemostat cultures grown at a constant dilution rate of 0.055 h(-1) was studied. Xylitol was the major product detected amongst all tested conditions. The maximal values for xylitol yield and volumetric pr...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(00)00166-6
更新日期：2000-06-01 00:00:00
abstract：:beta-Xylosidase (1,4-beta-D-xylan xylohydrolase EC 3.2.1.37) and xylose isomerase (D-xylose ketol-isomerase EC 5.3.1.5) produced by Streptomyces sp. strain EC 10, were cell-bound enzymes induced by xylan, straw, and xylose. Enzyme production was subjected to a form of carbon catabolite repression by glycerol. beta-Xyl...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(00)00182-4
更新日期：2000-07-01 00:00:00
abstract：:Detection of mRNA is a valuable method for monitoring the specific gene expression. In this study, we devised a novel cDNA synthesis method using three enzymes, the genetically engineered thermostable variant of reverse transcriptase (RT), MM4 (E286R/E302K/L435R/D524A) from Moloney murine leukemia virus (MMLV), the ge...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2016.10.003
更新日期：2017-01-01 00:00:00
abstract：:Enzymatic synthesis of cephalothin from 7-aminocephalosporanic acid (7-ACA) and amide derivatives of 2-thienylacetic acid (2-TA) using penicillin G acylase (pen G acylase) was studied. Two amide derivatives of 2-TA namely 2-thienylacetamide (2-TAA) and 2-thienylacetohydroxamic acid (2-TAH) were used in this study. The...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(99)00153-2
更新日期：2000-02-01 00:00:00
