Abstract：
:Zymomonas mobilis genes encoding INVA and INVB were expressed in Pichia pastoris, under the control of the strong AOX1 promoter, and the recombinant enzymes were named INVAAOX1 and INVBAOX1. The expression levels of INVAAOX1 (1660 U/mg) and INVBAOX1 (1993 U/mg) in P. pastoris were 9- and 7-fold higher than those observed for the native INVA and INVB proteins in Z. mobilis. INVAAOX1 and INVBAOX1 displayed a 2- to 3-fold higher substrate affinity, and a 2- to 200-fold higher catalytic efficiency (kcat/KM) than that observed for native INVA and INVB from Z. mobilis. Positive Schiff staining of INVAAOX1 and INVBAOX1 suggested a glycoprotein nature of both invertases. After deglycosylation of these enzymes, denoted D-INVAAOX1 and D-INVBAOX1, they exhibited a 1.3- and 3-fold lower catalytic efficiency (107 and 164 s(-1) mM(-1), respectively), and a 1.3- to 5-fold lower thermal stability than the glycosylated forms at temperatures of 35-45 °C. After deglycosylation no effect was observed in optimal pH, being of 5.5 for INVAAOX1, INVBAOX1, D-INVAAOX1 and D-INVBAOX1. The invertase activity of both enzymes increased in 80% (INVAAOX1) and 20% (INVBAOX1) in the presence of Mn(2+) at 1 mM and 5 mM, respectively. INVAAOX1 and INVBAOX1 were highly active at sucrose concentrations of up to 400 and 300 mM, respectively; however, the tolerance to sucrose decreased to 300 mM for D-INVAAOX1. Our findings suggest that glycosylation of INVAAOX1 and INVBAOX1 plays an important role in their thermal stability, catalytic efficiency, and tolerance to sucrose. In conclusion, the expression of INVA and INVB from Z. mobilis in P. pastoris yields new catalysts with improved catalytic properties, making them suitable candidates for a number of industrial applications or for the improvement of ethanol production from cane molasses.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Pérez de los Santos AI,Cayetano-Cruz M,Gutiérrez-Antón M,Santiago-Hernández A,Plascencia-Espinosa M,Farrés A,Hidalgo-Lara MEdoi
10.1016/j.enzmictec.2015.11.008subject
Has Abstractpub_date
2016-02-01 00:00:00pages
48-56eissn
0141-0229issn
1879-0909pii
S0141-0229(15)30082-Xjournal_volume
83pub_type
杂志文章abstract：:Flocculent yeast Saccharomyces cerevisiae YF234 (MATa ura3-52 trp1Delta2 his ade 2-1 can1-100 sta1 FLO8) cells overexpressing glyoxalase I and having strong flocculation ability were permeabilized with isopropyl alcohol and ethanol under various conditions. The treatment with 40% isopropyl alcohol significantly improv...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(00)00304-5
更新日期：2000-12-01 00:00:00
abstract：:This study examined the potential of untreated and alkali-pretreated sugarcane bagasse (SCB) in cellulase, reducing sugar (RS) and fungal biomass production via solid state fermentation (SSF) using Pycnoporus sanguineus. The impact of the composition, structure and cellulase adsorption ability of SCB on the production...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2013.05.005
更新日期：2013-09-10 00:00:00
abstract：:Direct cellobiose production from cellulose by a genetically modified fungus-Neurospora crassa, was explored in this study. A library of N. crassa sextuple beta-glucosidase (bgl) gene deletion strains was constructed. Various concentrations of cellobiose were detected in the culture broth of the N. crassa sextuple bet...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2012.12.010
更新日期：2013-03-05 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：: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：:The present study investigated the influence of low-shear modeled microgravity (LSMMG) conditions on Listeria monocytogenes stress response (heat, cold, and acid), membrane fatty acid composition, and virulence potential as well as stress-/virulence-associated gene expression. The results showed that LSMMG-cultivated ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2019.109440
更新日期：2020-02-01 00:00:00
abstract：:The parameters of biosensors based on the yeast strains Candida maltosa VKM Y-2359, Candida blankii VKM Y-2675, and Debaryomyces hansenii VKM Y-2482 for biochemical oxygen demand (BOD) detection are compared. The catalytic activity of the strains was analyzed in relation to the growth phase. The possibility of using D...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2012.01.002
更新日期：2012-04-05 00:00:00
abstract：:Different whole-cell immobilization methods and conditions were tested for the delta 1-dehydrogenation of 6-alpha-methyl-hydrocortisone-21-acetate with Arthrobacter simplex cells, in n-decane-1-ol and chloroform. Among the entrapment methods, polyurethane foams gave the best productivity of n-decane-1-ol, but could no...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(92)90036-n
更新日期：1992-08-01 00:00:00
abstract：:A chemiluminescence fiber optic system coupled to flow injection analysis (FIA) and ion exchange chromatography has been developed for determining glucose in blood and urine. Immobilized glucose oxidase acted on beta-D-glucose to produce hydrogen peroxide, which was then reacted with luminol in the presence of ferricy...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(93)90130-t
更新日期：1993-05-01 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：:Foaming, a proficient method for the recovery of surface active solutes from dilute solutions, was successfully applied for the concentration of the lipopeptide biosurfactant Surfactin from B. subtilis ATCC 21332 cell culture broths. Foaming was only partially successful in concentrating Surfactin when applied as a se...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(00)00327-6
更新日期：2001-03-08 00:00:00
abstract：:When dissolved oxygen (DO) was maintained at saturation level during batch fermentations of Streptomyces clavuligerus (NRRL 3585), the accumulation of the intermediate penicillin N was lowered while formation of the end product cephamycin C was increased relative to fermentations without DO control. The specific activ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(90)90178-s
更新日期：1990-01-01 00:00:00
abstract：:NADPH is the key cofactor in L-isoleucine (Ile) biosynthetic pathway. To increase the Ile biosynthesis in Corynebacterium glutamicum ssp. lactofermentum JHI3-156, NADPH supply needs to be enhanced. Here NAD kinase, the key enzyme for the de novo biosynthesis of NADP(+) and NADPH, were cloned and expressed in JHI3-156,...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2012.04.003
更新日期：2012-07-15 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：: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：: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：: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：:The recombinant shuttle plasmid pCPPS-31, which confers carboxymethylcellulase (CMCase) production and neomycin resistance (Ner), was segregationally but not structurally unstable in both B. subtilis (50-60%) and Escherichia coli (80-90%) when grown in serial batch cultures for 72 h diluting 1:1,000 with fresh medium ...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(94)90049-3
更新日期：1994-03-01 00:00:00
abstract：:During toluene hydroxylation, catalyzed by Pseudomonas putida UV4 one molecule of oxygen is added to the aromatic ring to produce the dihydroxylated (non-aromatic) ring structure, toluene cis-glycol. Toluene, which is toxic to the cells at aqueous phase concentration above ( approximately 2.4 mmol), is fed to the reac...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/s0141-0229(00)00289-1
更新日期：2001-02-01 00:00:00
abstract：:Conventional lipase screening methods are mostly based on hydrolytic activity, which may not always be the best method to assess the enzyme activity, especially for evaluating synthetic activity. Here we developed a high throughput and visual method to screen clones with high synthetic activity and used it to assess l...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2012.03.002
更新日期：2012-05-10 00:00:00
abstract：:We here describe a unique β-D-glucosidase (BGL; Blon_0625) derived from Bifidobacterium longum subsp. infantis ATCC 15697. The Blon_0625 gene was expressed by recombinant Escherichia coli. Purified recombinant Blon_0625 retains hydrolyzing activity against both p-nitrophenyl-β-D-glucopyranoside (pNPG; 17.3±0.24Umg(-1)...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2014.10.001
更新日期：2015-01-01 00:00:00
abstract：:Global substitution of canonical amino acids (cAAs) with noncanonical (ncAAs) counterparts in proteins whose function is dependent on post-translational events such as cofactor binding is still a methodically challenging and difficult task as ncAA insertion generally interferes with the cofactor biosynthesis machinery...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2017.06.014
更新日期：2017-11-01 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：:Horseradish peroxidase (HRP) was successfully purified from horseradish roots by a two-stage reverse-micellar extraction from the dialyzed aqueous extract. The anionic surfactant AOT dissolved in isooctane was used to produce the reverse-micellar phases. The narrow pH range at which HRP solubilization occurred was exp...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(95)00031-3
更新日期：1996-04-01 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：:A novel magnetic Fe3O4 nanoparticles (MNPs) coupled with agarose (AMNPs) was synthesized using co-precipitation via alkaline condition and span-80 surfactants in organic solvent. Iminodiacetate was first attached to the MNPs through epichlorohydrin agent and then chelated with metal ions. The morphology and chemical p...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2014.05.008
更新日期：2014-09-01 00:00:00
abstract：:Water-insoluble nucleases were prepared by immobilizing the endonuclease from S. aureus onto the surface of nylon-66 and polystyrene spheres. The activation phase of the synthetic supports was optimized to define optimal conditions of pH, temperature, and Ca2+ concentration for using immobilized enzymes. The activity,...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(93)90140-w
更新日期：1993-03-01 00:00:00
abstract：:Genetic engineering has been used to modify the proportion of different cellulases produced by a hypercellulolytic Trichoderma reesei mutant strain. A general expression vector, pAMH110, containing the promoter and terminator sequences of the strongly expressed main cellobiohydrolase 1 (cbh1) gene was used to overexpr...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/0141-0229(91)90133-u
更新日期：1991-03-01 00:00:00
abstract：:In this work, porous biochar derived from sugarcane bagasse was prepared and then coated with different amounts of chitosan (C@CS) for cellulase immobilization. Cellulase was covalently immobilized on the support by using glutaraldehyde as a linker. The chemical characteristics and morphology of the samples were deter...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2020.109561
更新日期：2020-08-01 00:00:00
abstract：:The xylanase R8 gene (xynR8) from uncultured rumen fungi was cloned and successfully expressed in Lactobacillus reuteri. A xylanase activity of 132.1 U/mL was found in the broth of L. reuteri R8, the transformant containing pNZ3004 vector with xynR8 gene insertion. Two distinct forms of recombinant xylanase with diffe...
journal_title：Enzyme and microbial technology
pub_type： 杂志文章
doi：10.1016/j.enzmictec.2014.06.004
更新日期：2014-10-01 00:00:00
