Production method of trehalose

A technology of trehalose and Pseudomonas putida, applied in the fields of fermentation engineering and microorganisms, can solve the problems of restricting the wide application of trehalose, high cost and low yield

Active Publication Date: 2010-09-15
ZHUCHENG DONGXIAO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0021] Currently commercialized trehalose is mainly extracted from yeast cells, th

Method used

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  • Production method of trehalose
  • Production method of trehalose

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043]Pseudomonas putida was collected and fermented on a shaking table at 30°C and 180r / min for 72h. The fermentation medium is: maltose 2wt%, peptone 0.5wt%, beef extract 0.2wt%, K 2 HPO 4 0.1wt%, NaH 2 PO 4 0.1wt%, MgSO 4 0.05wt%, balance water. After fermentation and cultivation, centrifuge at 4800r / min for 20min to obtain 100mL of concentrated bacterial liquid, which contains 10g of bacterial cells. Add 0.5mL of β-mercaptoethanol to the above-mentioned concentrated bacterial solution, and react for 2h to obtain a permeable cell solution of Pseudomonas putida. After 12 hours of centrifugation, the supernatant was taken to obtain a trehalose solution containing 4 g of trehalose.

Embodiment 2

[0045] Pseudomonas putida was collected and fermented on a shaking table at 30°C and 180r / min for 72h. The fermentation medium is: maltose 2wt%, peptone 0.5wt%, beef extract 0.2wt%, K 2 HPO 4 0.1wt%, NaH 2 PO 4 0.1wt%, MgSO 4 0.05wt%, balance water. After fermentation and cultivation, centrifuge at 10000r / min for 5min to obtain 50mL of concentrated bacterial liquid, which contains 5g of bacterial cells. Add 0.5mL of β-mercaptoethanol to the above-mentioned concentrated bacterial solution, and react for 2h to obtain a permeable cell solution of Pseudomonas putida, then add 5g of maltose to the obtained permeable cell solution of Pseudomonas putida, and react After 12 hours of centrifugation, the supernatant was taken to obtain a trehalose solution containing 2 g of trehalose.

Embodiment 3

[0047] Pseudomonas putida was collected and fermented on a shaking table at 30°C and 180r / min for 72h. The fermentation medium is: maltose 2wt%, peptone 0.5wt%, beef extract 0.2wt%, K 2 HPO 4 0.1wt%, NaH 2 PO 4 0.1wt%, MgSO 4 0.05wt%, balance water. After fermentation and cultivation, centrifuge at 8000r / min for 10min to obtain 1000mL of concentrated bacterial liquid, which contains 80g of bacterial cells. Add 8 mL of β-mercaptoethanol to the above-mentioned concentrated bacterial liquid, react for 2 hours, and prepare a permeable cell solution of Pseudomonas putida, then add 100 g of maltose to the prepared permeable cell solution of Pseudomonas putida, and react for 12 hours , after centrifugation, the supernatant was taken to obtain a trehalose solution containing 31g of trehalose. After thin-layer chromatography of the trehalose solution and the untreated Pseudomonas putida cell solution cultured under the same conditions, it was found that about 30% of the maltos...

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Abstract

The invention relates to a production method of trehalose, in particular to a method for producing trehalose by utilizing pseudomonas putida, belonging to the technical field of fermentation engineering and microorganism. The method for producing trehalose by utilizing pseudomonas putida comprises the following steps of: culturing the pseudomonas putida, treating beta-mercaptoethanol and fermenting. In the method, maltose is used as a substrate, and then the maltose is transformed to the trehalose in one step through preparing the permeable cells of pseudomonas putida by utilizing trehalose synthase in the cells of the pseudomonas putida. Through determination, about 3-5% of trehalose is generated in the permeable cell solution of the pseudomonas putida treated by using the beta-mercaptoethanol.

Description

technical field [0001] The invention relates to a method for producing trehalose, in particular to a method for producing trehalose by using Pseudomonas putida, which belongs to the field of fermentation engineering and microorganism technology. Background technique [0002] Trehalose (Trehalose), also known as mushroom sugar, Qin sugar, its molecular formula is C 12 h 22 o 11 2H 2 O, the chemical name α-D-glucopyranosyl α-D-glucopyranoside (α-D-glycopyranosyl-α-D-glycopyranoside), is a non-glucoside formed by condensation of two glucose molecules through hemiacetal hydroxyl Reducing disaccharides. Widely present in bacteria, yeast, fungi, algae and insects. Trehalose has unique biological functions, which can protect proteins, biofilms and cell walls of sensitive cells from damage caused by drought, freezing and changes in osmotic pressure. It can be used as a stabilizer for unstable drugs, food and cosmetics, and a variety of Food and pharmaceutical sweeteners, seed ...

Claims

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Application Information

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IPC IPC(8): C12P19/12C12R1/40
Inventor 王瑞明马春玲王腾飞李丕武徐汝意
Owner ZHUCHENG DONGXIAO BIOTECH
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