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Meyerozyma guilliermondii and application of secreted extracellular protein

An extracellular protein and yeast technology, applied in the field of microorganisms, to achieve the effects of mild reaction conditions, good specificity and high catalytic efficiency

Active Publication Date: 2020-10-16
CHENGDU BIOPURIFY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no public reports on the preparation and production of simenoside I in large quantities

Method used

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  • Meyerozyma guilliermondii and application of secreted extracellular protein
  • Meyerozyma guilliermondii and application of secreted extracellular protein
  • Meyerozyma guilliermondii and application of secreted extracellular protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of extracellular protein produced by Pichia pastoris (Meyerozyma guilliermondii) simonoside I

[0044] Step 1: Pichia guilliermondii (Meyerozyma guilliermondii) was streaked and inoculated on a fresh slant medium, and cultured at 25° C. for 48 hours to obtain activated slant strains;

[0045] Step 2: Inoculate the activated slant strains obtained in step 1 in a 100mL Erlenmeyer flask with 20mL of seed medium, and culture it on a shaker at 25°C and 200rpm for 48h to obtain the seed solution; v) Inoculation amount of -10% (v / v), inoculate the seed solution in a 500mL Erlenmeyer flask equipped with 200mL liquid fermentation medium, culture on a shaking table at 25°C and 200rpm for 48h, to obtain a fermentation broth;

[0046] Step 3: centrifuging the fermented liquid obtained in step 2, collecting the cell precipitate and supernatant, and concentrating the supernatant to obtain the extracellular protein;

[0047] Wherein in step 1, the slant medium i...

Embodiment 2

[0050] Example 2 Preparation of simonoside I by using the extracellular protein of Luo Han Guo endophyte LHGNSJ-VS01

[0051] 5g / L crude LHGNSJ-VS01 extracellular protein, 10g / L mogroside V, mixed with 0.2M phosphate buffer pH=7.2, stirred at 25°C and 200rpm, reacted for 24h, and the substrate conversion rate reached 31%. Symonoside I The purity is 98.5%, as figure 2 shown. For symmonoside I spectrum data, see Figure 5 and Image 6 .

Embodiment 3

[0052] Example 3 Preparation of simonoside I by using the extracellular protein of Luo Han Guo endophyte LHGNSJ-VS01

[0053] 1g / L crude LHGNSJ-VS01 extracellular protein, 5g / L mogroside V, 0.2M phosphate buffer pH=7.2, 25°C, 200rpm, react for 24h. The substrate conversion rate reaches 31%, and the purity of simonoside I is 98.5%, such as Figure 4 shown.

[0054] The assay method of Luo Han Guo extract content in above-mentioned embodiment 2 or 3:

[0055] High performance liquid chromatography (HPLC) was used to determine the content of simonoside I and other Luo Han Guo extracts.

[0056] Sample treatment: Add the same volume of ethanol to the reaction solution to precipitate the protein, and dissolve the substrate (mogroside V) and the product

[0057] Substance (Simanoside I), centrifuged at 10000r / min for 6min, filtered through a 0.45μm filter membrane, to be tested.

[0058] Chromatographic column: Ultimate○RXB-C18 column (150mm×4.6mm×3μm, );

[0059] Mobile phas...

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Abstract

The invention discloses Meyerozyma guilliermondii and an application of secreted extracellular protein, relates to the technical field of microorganisms, and particularly relates to a method for efficiently synthesizing siamenoside I by using the Meyerozyma guilliermondii. According to the application, the catalytic function is achieved through the extracellular protein secreted by the Meyerozymaguilliermondii; the extracellular protein has relatively strong catalytic activity, has the characteristics of high catalytic efficiency, mild reaction conditions, strong specificity and the like, isexpected to be an ideal catalyst for efficiently synthesizing the siamenoside I, and widens the source of the siamenoside I.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a method for efficiently synthesizing simonoside I by using Pichia mongolica. Background technique [0002] Luo Han Guo (Siraitia grosvenorii) is an authentic medicinal material in Guangxi. It has strict requirements on the habitat. It is mainly distributed vertically in subtropical slope areas at an altitude of 250-1000m. There are many plants such as moss, orchids, camellia oleifera and bamboo forests in the distribution area; Luo Han Guo It is required to grow in areas with large temperature difference between day and night and humid environment. The optimum temperature is 25-30°C, and the relative air humidity is 70%-85%. It likes light but is not resistant to strong light and is afraid of frost. Rich, loose and moist red-yellow loam. Lingui, Yongfu, Xing'an, Ziyuan, Longsheng, and Lingchuan in Guilin have the largest cultivation areas and are also the largest product...

Claims

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Application Information

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IPC IPC(8): C12P33/20C12P21/00C12R1/645
CPCC12P33/20C12P21/00Y02P20/584
Inventor 王鸿江谢海峰谢期林胡云岭
Owner CHENGDU BIOPURIFY LTD
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