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Microbacterium barkeri XJ and method for preparing steviol by microbacterium barkeri XJ

A technology of Microbacillus pasteurii and steviol, which is applied in the field of steviol glycoside or rubus glycoside conversion by microorganisms or enzymes, and the preparation of steviol can solve the problems of limited source of helicase and high price

Active Publication Date: 2011-10-19
NANJING NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Steviol can be obtained by treating steviol glycosides with helicase and toluene. When degrading steviol glycosides, helicase acts on the 1-4 glycosidic bond in an endo-cutting manner, and the reaction is complete in 48 hours. After separation and purification, steviol can be obtained (Liu Zhujin, Zhou Wenhua, Gao Feng , et al. Study on the Sweetness Components of Rubus cotyledons. Botany Bulletin, 1983, 01: 33-37), but the source of helicase is limited and the price is high, and the toluene used is a toxic organic solvent

Method used

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  • Microbacterium barkeri XJ and method for preparing steviol by microbacterium barkeri XJ
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  • Microbacterium barkeri XJ and method for preparing steviol by microbacterium barkeri XJ

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preparation example Construction

[0034] Preparation of Microbacterium pasteurianum powder or enzyme powder: Use conventional liquid or solid fermentation culture and enzyme separation methods to obtain bacteria powder or enzyme powder.

[0035] For example, the preparation of bacterial powder includes fermentation of bacteria, separation of fermentation broth, emulsification of bacterial sludge and vacuum freeze-drying of emulsion. The enzyme powder fermented by the enzyme produced by Microbacterium pasteurianus is frozen and then vacuum-dried for 30 h at a vacuum degree of 1-5 Pa.

[0036] The material used in the present invention is the transformation substrate: steviol glycoside and rubusoside can be purchased from Changsha Guangming Food Chemical Company, and can also be prepared from stevia rebaudiana or rubus cotyledons by common extraction methods.

Embodiment 1

[0039] Prepare 1% (W / W), pH 7.5 steviol glycoside conversion substrate (steviol glycoside: water = 1::99 (W / W), the preparation method is the same below), insert 1% (V / V) Pasteur Microbial The bacillus liquid was cultured dynamically at 37°C and 220 r / min for 3 days. After the cultivation, the bacteria were removed by centrifugation at 5000 rpm. The supernatant was flocculated and precipitated to remove impurities, the macroporous resin was adsorbed and separated, and the eluent was crystallized and purified to obtain high-purity steviol with a purity of about 95%. The conversion rate of stevioside was 90%.

Embodiment 2

[0041] 1% (W / W), pH 6.5 steviol glycoside substrate in a 500 mL transformation bottle, add 10% (W / W) enzyme solution, and transform at 37°C for 2 days. After separation and purification, steviol with a purity of about 95% is obtained.

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Abstract

The invention relates to microbacterium barkeri XJ strain and application thereof. The preservation number of microbacterium barkeri XJ is CCTCC NO.M2011182. The strain can be used for preparing steviol, particularly, the method comprises the following steps: preparing stevioside or rubusoside into bacterium culture solution or enzymatic conversion solution; adding a proper quantity of bacterium or enzyme, then conversing stevioside or rubusoside into steviol under the conditions of proper conversion liquid quantity, conversion temperature, pH, time and the like; and separating and purifying the conversion solution to obtain high-purity steviol. The invention also provides the method for effectively preparing steviol.

Description

technical field [0001] The invention relates to a new method for preparing steviol, in particular to a method for obtaining steviol by converting steviol glycoside or rubusoside with microorganisms or enzymes. The invention belongs to the field of production and application of pharmaceutical raw materials. Background technique [0002] Stevioside is a white, odorless powder, slightly soluble in ethanol, stable to heat, acid, and alkali, and the solubility at room temperature exceeds 40% (Wang Yifan, Ren Dai, Zhou Fengxian, etc. Research on nutritional components in stevioside foods. Modern China Journal of Medicine, 1997, 7(3): 63-64). [0003] Stevioside is a mixture of eight diterpene glycosides. The general structural formula is shown in Table 1. The R1 and R2 positions can be connected with different types and quantities of monosaccharides to form different components. The eight components are: stevioside (glycoside, the following glycoside Synonymous with glycoside), ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P15/00C12R1/01
Inventor 陈育如玄燕赵乙萱刘润薇
Owner NANJING NORMAL UNIVERSITY
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