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Supervisory control method for producing trehalose by double-enzyme method

A technology of trehalose and double-enzyme method is applied in the field of monitoring and control of trehalose production by double-enzyme method, and achieves the effects of saving labor, reducing monitoring and production costs, and achieving accurate and reliable results.

Active Publication Date: 2017-12-08
CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to change the DE value detection and trehalose content detection which need to be manually frequently carried out in the process of trehalose production by double enzyme method to near-infrared automatic detection, and automatically control the amount of alkali added according to the change to realize production optimization. The purpose of automatic regulation is to provide a method: apply near-infrared real-time detection of DE value and trehalose content of starch liquefaction liquid in the process of double-enzyme production of trehalose, and establish the relationship between the change of its content and the change of pH value through calculation and analysis Model and correct, and then determine the time and amount of alkali addition to stabilize the pH value of the reaction solution so that it is in the optimum pH range with the highest activity of MTSase and MTHase, so as to achieve the purpose of automatic regulation of production optimization

Method used

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  • Supervisory control method for producing trehalose by double-enzyme method
  • Supervisory control method for producing trehalose by double-enzyme method
  • Supervisory control method for producing trehalose by double-enzyme method

Examples

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Effect test

Embodiment 2

[0019] Example 2 Starch liquefaction and detection: The starch and water are adjusted into starch milk at a weight ratio of 20:100 in the storage tank (1), pH is adjusted to 5.5, and high temperature resistant is added according to the weight ratio of enzyme: starch = 2:5000 α-Amylase is transported to the jet liquefier (3), heated to 90°C through the steam inlet pipe (4), and maintained in the liquefaction tank (6) for 60 minutes to liquefy the starch. Time-sharing sampling is performed every 10 minutes. The DE value is measured by Fehling's reagent method, and the near-infrared spectrum corresponding to each sample is collected through the optical fiber (9) and the near-infrared spectrometer (10).

Embodiment 3

[0020] Example 3 Starch dextrin cutting branches: take the liquefied solution with DE value of 15 and add HCI to the reaction tank (13) to adjust the pH to 2 and keep the enzyme for 15 minutes, then adjust NaOH to adjust the pH to 5.6, press enzyme: starch = 2 : 1000 weight ratio plus pullulanase to cut the amylopectin into amylose dextrin.

Embodiment 4

[0021] Example 4 Catalytic reaction and detection: Adjust the temperature of the liquefied liquid after cutting branches in Example 3 to 44-46°C, add acid or alkali to adjust the pH to 6, according to MTSase: MTHase: starch dextrin = 4:8:1000 The weight ratio is added from Rhizobium ( Rhizobium sp. ) The MTSase and MTHase ​​enzyme solutions of CCTCC AB207877 respectively took time-sharing sampling of the two catalytic reactions without pH adjustment and pH adjustment during the catalytic reaction process, and measured the pH value of each sample in real time with a Shanghai Lemag pH meter; using high performance liquid chromatography Method for real-time determination of the trehalose content of each sample, the results are shown in Table 2. HPLC conditions: amino column-Agela InnovalNH 2 , 5 μm, 4.6×250 mm; mobile phase: acetonitrile / water, volume ratio 80 / 20; flow rate: 1.0 mL / min; detector: parallax refractive index detector; injection volume: 10 μL; column temperature: 35 ...

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Abstract

The invention discloses a supervisory control method for producing trehalose by a double-enzyme method. The supervisory control method comprises the following steps of firstly mounting a near-infrared real-time monitoring and control system; collecting starch liquefied liquid, measuring a DE (Dextrose Equivalent) value by using a Fehling's reagent method, collecting catalysis liquid, measuring a pH (potential of Hydrogen) value by using a pH meter, measuring the content of the trehalose by using high performance liquid chromatography, associating with a correspondingly collected near-infrared spectrum, establishing a quantitative model of changes of the DE value, the content of the trehalose and the pH value in a production process, and correcting; afterwards, quickly detecting the DE value, the content of the trehalose and the pH value in the production process by utilizing the established model, and stabilizing the DE value to be 8 to 20 and stabilizing the pH value to be in the range, in which the activity of malto-oligosaccharide based trehalose synthase-MTSase and malto-oligosaccharide based trehalose hydrolase-MTHase is optimal, by utilizing a control system. The method can be used for monitoring the DE value of starch liquid and the content of the trehalose in reaction liquid in real time and can also used for automatically controlling the change of the pH value and the content of the trehalose and the result is accurate and reliable.

Description

Technical field [0001] The present invention belongs to an invention related to trehalose. More specifically, the present invention relates to a method for monitoring and controlling the production of trehalose by a two-enzyme method. Background technique [0002] Trehalose-Trehalose is a non-reducing disaccharide composed of two molecules of glucose connected by α,α-1,1-glycosidic bonds. It is known as "Trehalose" because of its excellent resistance to stress and protection against organisms. "Sugars of life" and "new sugars in the 21st century", their large-scale manufacturing methods have attracted widespread attention; the production methods of trehalose include natural biological extraction, microbial fermentation, chemical synthesis, genetic engineering and enzyme conversion Method; the current industrial production basically adopts the enzyme conversion method, mainly divided into single-enzyme method and double-enzyme method; in the production process of trehalose by doub...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12P19/12C12Q3/00
CPCC12P19/12C12P19/14C12Q3/00
Inventor 谢定杨倩圆郑瑞娜谢易真
Owner CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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