Paenarthrobacter ilicis CR5301 and application thereof

A technology for biotransformation and stevioside, applied in microorganisms, bacteria, biochemical equipment and methods, etc., can solve the problems of long conversion time, troublesome separation and purification, and difficult conversion of rebaudioside C.

Active Publication Date: 2021-11-19
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From the current research, the strains that can use stevioside for microbial transformation mainly include Sulfolobus solfataricus, Penicillium decumbens, Aspergillus aculeatus, Penicillium pinophilum, Microbacterium barkeri and human feces Isolated bacterial strains, etc. Most of these microorganisms use stevioside in stevioside as a substrate, only a small number of microorganisms can hydrolyze rebaudioside A, and fewer strains can transform rebaudioside C, and the transformation time is also shorter. It is very long, because the glycosyl groups connected to C19 and C13 of rebaudioside C lead to the difficulty of conversion of rebaudioside C, which brings great troubles to the subsequent separation and purification. The strains of diglycoside C, shortening the conversion time, reducing the cost of reusing by-products, and improving the application value of by-products are the key issues we need to solve

Method used

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  • Paenarthrobacter ilicis CR5301 and application thereof
  • Paenarthrobacter ilicis CR5301 and application thereof
  • Paenarthrobacter ilicis CR5301 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Isolation and preservation of Paenarthrobacter ilicis CR5301

[0032] 1. Paenarthrobacter ilicis CR5301 isolation and transformation experiments:

[0033] The strain screening process is to select 10 soil samples from the stevia planting area in Hailin City, Heilongjiang Province, take 5g of each sample, add 45mL sterile saline, place on a shaker at 28°C, 135r / min, and shake for 30min. Dilute the sample solution to 10 -2 -10 -6 , spread the dilution on the plate separation medium, carry out 3 repeated experiments for each dilution, culture in a 28°C incubator for 3-5 days, observe the growth of the colony on the plate, pick a single colony, and inoculate it in the activated Culture medium, after 2 days of culture at 28°C, diluted again and spread on the plate separation medium for re-screening, picked a single colony and inoculated it in the activation medium, cultured at 28°C for 2 days, stored in a -80°C refrigerator with glycerol. After the re-screened ...

Embodiment 2

[0040] Example 2: Paenarthrobacter ilicis CR5301 is used for biotransformation of high-purity rebaudioside C.

[0041] Take out the CR5301 strain from the -80°C refrigerator, and inoculate it in the activation medium with 1% inoculum amount. When the OD600 value of the bacterial solution reaches about 1.0, take 8ml, centrifuge, add the bacteria to 20ml transformation medium, and inoculate at 28°C , 270r / min shaking culture for 8h, the conversion rate of rebaudioside C was 100%.

Embodiment 3

[0044] Example 3: Paenarthrobacter ilicis CR5301 is used for biotransformation of high-purity rebaudioside C.

[0045] Take out the CR5301 strain from the -80°C refrigerator, and inoculate it in the activation medium with 1% inoculation amount. When the OD600 value of the bacterial solution reaches about 1.0, take 8ml, centrifuge, add the bacteria to 20ml transformation medium, and inoculate at 20°C. , 270r / min shaking culture for 24h, the conversion rate of rebaudioside C was 74%.

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Abstract

The invention relates to Paenarthrobacter ilicis CR5301 and application of the Paenarthrobacter ilicis CR5301, and the preservation number of the Paenarthrobacter ilicis CR5301 is CCTCC (China Center for Type Culture Collection) NO: M 2021851. The Paenarthrobacter ilicis CR5301 is used for converting thalli of rebaudioside C in stevioside, the rebaudioside C which is bitter and astringent in taste and difficult to hydrolyze can be converted, the conversion speed is high, conversion substrates are not limited to the rebaudioside C, the Paenarthrobacter ilicis CR5301 can also convert stevioside, rebaudioside A, dulcoside A and other stevioside in stevioside byproducts, the conversion products are easy to purify, final products are all steviol, and the problem that by-products in a stevioside factory are difficult to utilize is solved.

Description

1. Technical field [0001] The present invention has discovered a Paenarthrobacter ilicis CR5301 for the first time, and the Latin classification of this bacterial strain is named as: Paenarthrobacter ilicis , the strain has been deposited in the China Center for Type Culture Collection on July 9, 2021. The depository unit is referred to as CCTCC, address: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, and the deposit number: CCTCC NO: M 2021851. [0002] The invention relates to strains for microbial transformation using stevioside, in particular to Paenarthrobacter ilicis CR5301 and applications thereof. 2. Background technology [0003] Stevioside is a calorie-free, high-sweet tetracyclic diterpenoid compound extracted from stevia rebaudiana. The sweetness of stevioside is 150-300 times that of sucrose. The main components are stevioside (ST) and rebaudioside A (RA) and Rebaudioside C (RC), and some components with very low content, such as Duke A, Reba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P33/00C12R1/01
CPCC12N1/20C12P33/00
Inventor 李洪飞曹龙奎孙大庆王宝辉
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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