Mọi người ơi, dịch giúp mình đoạn văn này với, nó là từ ngữ chuyên ngành, có những từ gần như mình bó tay, làm thế nào cũng không thể dịch hợp với ngữ cảnh được, khó khăn quá, hixhix.....thanks all!
Ellipsometry and Adsorption Experiments. Ellipsometric
measurements were performed using a vertical computer-controlled
DRE-EL02 ellipsometer (Ratzeburg, Germany). The angle of
incidence was set at 70.0°, and the wavelength, λ, of the He-Ne
laser was 632.8 nm. For the data interpretation(giải thích), a multilayer (nhiều tầng) model
composed(bao gồm) of the substrate( phức hợp, the unknown layer, and the surrounding
medium should be used. The thickness (dk) and refractive (có tính khúc xạ) index(chỉ số) (nk)
of the unknown layer can then be calculated from the ellipsometric
angles, ∆ and Ψ, using the fundamental ellipsometric equation and
iterative calculations with Jones matrices:19
ei∆ tan Ψ)Rp⁄Rs)f(nk, dk,λ,φ)
where Rp and Rs are the overall reflection coefficients for the parallel
and perpendicular waves, respectively. They are a function of the
angle of incidence φ, the wavelength λ of the radiation, the refractive
index (nk), and the thickness (dk) of each layer of the model.
From the ellipsometric angles ∆ and Ψ and a multilayer model
composed of silicon, silicon dioxide, enzyme layer, and air, it is
possible to determine only the thickness of the cellulase layer, d.
The thickness of the SiO2 layers was determined in air, considering
the refractive index for Si as n˜ ) 3.88 - i0.01820 and its thickness
as an infinite one; for the surrounding medium (air), the refractive
index was considered as 1.00. Because the native SiO2 layer is very
thin, its refractive index was set as 1.46220 and just the thickness
was calculated. The mean thickness of the native SiO2 layer amounted
to (1.9 ( 0.2) nm.
Adsorption kinetics was investigated either at (24 ( 1) °C or at
(60 ( 1) °C by immersing Si wafers into cellulase solution at the
concentration of 0.500 mg mL-1 during different periods of times,
which ranged from 0.5 to 72 h. After each period of time, the samples
were washed with an abundance of distilled water and dried under
a stream of N2. The mean thickness of adsorbed cellulase layers d
was determined in air, and the adsorbed amount of cellulase, Γ (mg
m-2), could be estimated by multiplying d by the density F of a dry
enzyme layer (F ∼ 1.37 g cm-3):21
Γ ) Fd
Adsorption isotherms of cellulase onto silicon wafers were
obtained by immersing the substrates into cellulase solutions
prepared in the concentration range of 0.005-1.00 g L-1 at pH
6.5, either at (24 ( 1) °C or at (60 ( 1) °C. The samples were then
washed with an abundance of distilled water and dried under a stream
of N2, and then the mean thickness of adsorbed cellulase layers d was
determined in air and Γ was calculated using eq 2. Cellulase desorption
was investigated at (24 ( 1) °C or at (60 ( 1) °C by monitoring
cellulase adsorbed layer thickness after immersion in distilled water
during 24 h.
Atomic force microscopy (AFM) topographic images were
obtained using a PicoSPM-LE Molecular Imaging system with
cantilevers operating at intermittent-contact mode (AAC mode), in
their resonance frequency of approximately 327 kHz in air. All
topographic images represent unfiltered original data and refer to
scan areas ranging from 1.3 µm × 1.3 µm to 2.0 µm × 2.0 µm with
a resolution of (512 × 512) pixels. At least two samples of the same
material were analyzed at different areas of the surface. Image
processing and determination of mean roughness (rms) were
performed by using the PicoScan 5.3.2 software.
Catalytic ActiVity of Adsorbed Cellulase and Free Cellulase. The
hydrolysis of cellulose catalyzed by cellulase yields glucose as the
product (see Scheme 1 in the Supporting Information). In this work,
glucose was quantified using coupled reactions with hexokinase
type III, from Baker's yeast (HK, H5000), and glucose-6-phosphate
dehydrogenase, also from Baker's yeast (G-6-PDH, G4134):
The reduction of nicotinamide adenine dinucleotide phosphate
sodium salt (NADP+, N0505) to NADPH was monitored by UV-vis
spectrophotometry (Beckman-Coulter DU 600) as a function of time,
at the wavelength of 340 nm, according to Bergmeyer's method.22
The concentration of NADPH was calculated using the Lambert-Beer
equation and considering the optical pathway as 1 cm and molar
extinction coefficient23 of NADPH of 6.220 L mol-1 cm-1. The final
value of NADPH concentration was measured after 0.5 h reaction,
since absorbance values were constant already after 20 min. Therefore,
NADPH concentration is directly proportional to glucose formed by
hydrolytic activity of cellulase.
In order to test the hydrolysis of cellulose catalyzed by immobilized
cellulase, six pieces of silicon wafers, with a total of 6 cm2 of cellulase,
were immersed into a glass vial containing 10 mL of a 5 w/w %
cellulose dispersion prepared in acetate buffer (50 mmol L-1) at pH
4.6. The temperature was kept at 37, 45, or 60 °C during 24 h. The
recipe was just occasionally shaken, because continuous stirring
showed no influence on the catalytic activity. After 24 h, an aliquot
was collected and centrifuged for 4 min at 13 400 rpm. The
supernatant was removed, and the glucose content in it was quantified
by NADPH formation. For comparison, the catalytic activity of free
cellulase was also tested. For this, the amount of free cellulase used
was the quantity corresponding to that of immobilized cellulase,
which was added to 10 mL of cellulose dispersion, and, as described
above, NADPH formation was monitored. For the comparative
experiments, first the adsorbed amount (mass/area) was determined
by means of ellipsometry. Since the Si wafer area is known, the
corresponding mass of adsorbed cellulase can be estimated. Once
the volume of the cellulose dispersion is known, where the cellulase
covered Si wafers are immersed, the cellulase concentration can
be estimated. This estimated value of cellulase concentration was
used for free cellulase.
dangvanchung
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