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A method for modifying gold chips in situ with cellulase

A cellulase, in-situ modification technology, applied in the field of instrumental analysis, can solve problems such as unsustainability, scale limitation, and international food prices soaring

Active Publication Date: 2021-02-09
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the current industrial production of fuel ethanol technology is the first-generation bioethanol technology based on food crops, which is limited in scale and unsustainable in the long run, and may also cause a sharp rise in international food prices

Method used

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  • A method for modifying gold chips in situ with cellulase
  • A method for modifying gold chips in situ with cellulase
  • A method for modifying gold chips in situ with cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The chip used is a QCM chip with a gold surface.

[0030] The first is the preparation of the gold chip: clean the gold chip and blow it dry with nitrogen, put it in a UV-ozone cleaner for 10-30 minutes, and then place the chip in the QCM flow cell.

[0031] Then start the in situ modification procedure:

[0032] Firstly, 75% ethanol solution was introduced at a rate of 0.1ml / min until the baseline became stable. Then a mixed solution of 40 mM 11-mercaptoundecanoic acid (MPA) and 40 mM 3-mercaptopropionic acid (MUA) mixed at a ratio of 10:1 (V:V) was injected at a rate of 0.1 ml / min. The QCM frequency drops to about -6Hz,. After equilibrium, rinse with 75% ethanol, and the frequency returns to about -4Hz. The energy dissipation value (D) of the whole process is almost inconvenient. Calculated according to the Sauerbrey equation, the adsorption amount of MPA / MUA on the gold chip is about 1.1mg / m 2 , the thickness of the film is about 0.6nm.

[0033] Next, 2 mM 1-et...

Embodiment 2

[0036] The chip used is an SPR chip with a gold surface.

[0037]The first is the preparation of the gold chip: clean the gold chip and dry it with nitrogen gas, put it in a UV-ozone cleaner for 10-30 minutes, and then place the chip in the SPR flow cell.

[0038] Then start the in situ modification procedure:

[0039] Firstly, 75% ethanol solution was injected at a rate of 0.1ml / min until the baseline became stable and the SPR angle was about 72.68°. Then a mixed solution of 40 mM 11-mercaptoundecanoic acid (MUA) and 40 mM 3-mercaptopropionic acid (MPA) mixed at a ratio of 10:1 (V:V) was injected at a rate of 0.1 ml / min. The SPR angle rises to about 72.89°. After equilibration, rinse with 75% ethanol at an almost inconvenient angle. According to the equation estimation, the adsorption thickness of MPA / MUA on the gold chip is about 3.8nm.

[0040] Next, 2 mM 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (NHSS) and 5 mM N-hydroxysulfosuccinyl mixed at a volume ...

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Abstract

The invention relates to a method for modifying a gold chip in situ with cellulase. Quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) technologies are important tools for real-time and in-situ research on the adsorption of biomacromolecules on solid interfaces. ) and the energy dissipation value of the adsorbed layer (corresponding to the structure of the thin film on the sensor), the latter only studying the change of "dry matter". The traditional method of studying cellulase and substrate is to immobilize the substrate on a QCM or SPR chip, and then pass cellulase as the mobile phase to study the interaction between the two. The present invention combines cellulase with a gold chip by in-situ modification method, constructs a cellulase film with a uniform surface, and broadens the application range of QCM or SPR to study the interaction between cellulase and other macromolecules in the system .

Description

technical field [0001] The invention relates to a method for in-situ modification of a gold chip by cellulase, belonging to the field of instrument analysis. Background technique [0002] According to the definition of the International Energy Agency (IEA), biomass refers to various organisms formed through photosynthesis, including all animals, plants and microorganisms. Biomass energy is the energy form of solar energy stored in biomass in the form of chemical energy. It has always been one of the important energy sources for human survival, and it is the fourth largest energy source after coal, oil, and natural gas. plays an important role in the system. [0003] In today's society, with the rapid development of the global economy, the demand for various energy sources is increasing day by day, and most of these energy sources are provided by coal mines and oil. On the one hand, the refining process of fossil fuels will bring some environmental pollution problems. On t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N29/02G01N5/00G01N21/552
CPCG01N5/00G01N21/553G01N29/022G01N2291/022
Inventor 宋君龙王沛沛杨益琴吴淑芳王志国任浩金永灿戴红旗
Owner NANJING FORESTRY UNIV
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