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Determination method for enzymatic activity of sucrose synthetase in cassava leaf

A technology for sucrose synthase and enzyme activity, which is applied in the field of measuring sucrose synthase enzyme activity, can solve the problems of complicated operation and long operation time, and achieves the effects of good reproducibility, strong comparability, and improved measurement sensitivity

Inactive Publication Date: 2013-11-20
GUANGXI SUBTROPICAL CROPS RES INST GUANGXI SUBTROPICAL AGRI PROD PROCESSING RES INST
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AI Technical Summary

Problems solved by technology

Sephadex G-25 column is used for the pretreatment of the enzyme solution to remove sugar. The elution and collection process is complicated and takes a long time. The operation process needs to be carried out in a low-temperature chromatography cabinet

Method used

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  • Determination method for enzymatic activity of sucrose synthetase in cassava leaf
  • Determination method for enzymatic activity of sucrose synthetase in cassava leaf
  • Determination method for enzymatic activity of sucrose synthetase in cassava leaf

Examples

Experimental program
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Embodiment 1

[0020] The method for measuring the activity of sucrose synthase in cassava leaves:

[0021] 1. Material collection and storage: Collect fresh cassava leaves and quickly put them into a low-temperature container. If the measurement cannot be carried out immediately, grind them with liquid nitrogen and store them in a -80°C low-temperature refrigerator for later use.

[0022] 2. Enzyme solution extraction: Take about 1g of pre-cooled cassava leaves, add a small amount of quartz sand and 5mL enzyme solution extraction buffer (100mmol / L, Tris-HCl with pH=7.2, 10mmol / L MgCl 2 , 1mmol / L EDTA-Na 2 , 10mmol / L β-mercaptoethanol, volume concentration 2% ethylene glycol, and weight concentration 1% polyvinylpyrrolidone (PVP) were quickly ground into a uniform paste in a mortar, cleaned the mortar, poured it into a centrifuge tube, and Centrifuge at 12000r / min for 15min in a low-temperature refrigerated ultracentrifuge, take the supernatant and discard the precipitate. The above extract...

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Abstract

The invention discloses a determination method for enzymatic activity of sucrose synthetase in a cassava leaf. The method comprises the following steps: determination of enzymatic activity of sucrose synthetase both in a synthesis direction and in a decomposition direction; and extracting enzyme liquid from a fresh cassava leaf, carrying out desugaring by using a dialysis bag and then determining enzymatic activity. Compared with the prior art, the invention has the following advantages: since dialysis desugaring treatment and enzymatic activity determination technology are combined together after extraction of the enzyme liquid, interference of high-background cane sugar content in the enzyme extract of the cassava leaf on determination of enzymatic activity of sucrose synthetase can be effectively eliminated, determination sensitivity is improved, determination process is easy to control, and determination results are reliable and have good comparability and reproducibility.

Description

technical field [0001] The invention relates to a method for measuring the activity of sucrose synthase, in particular to a method for measuring the activity of sucrose synthase in cassava leaves. Background technique [0002] Cassava stores a large amount of starch in the form of root tubers and is an important source of starch. Sucrose is an important product of photosynthesis and the main substance transported in the body. Sucrose synthase in cassava leaves is one of the key enzymes in the cassava starch synthesis process. Sucrose synthase can catalyze both sucrose synthesis and sucrose decomposition, and is one of the keys to control the strength and activity of the pool. A starchy substrate is sufficient. The determination of sucrose synthase activity in cassava leaves is affected by many factors. Because there is a large amount of photosynthetic product in cassava leaves—sucrose, which is much higher than the amount of sucrose produced in a short period of time durin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
Inventor 黄惠芳黄强侯学文罗燕春
Owner GUANGXI SUBTROPICAL CROPS RES INST GUANGXI SUBTROPICAL AGRI PROD PROCESSING RES INST
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