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Bacillus clausii and method for producing tetrahydropyrimidine by using same

A technology of ectoine and bacillus, applied in the field of microorganisms, can solve the problems of ectoine and other undiscovered, achieve good industrial application prospects, reduce fermentation equipment corrosion, and reduce the effect of separation and purification costs

Active Publication Date: 2020-06-19
BEIJING BIOINNO BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Bacillus clausii is a probiotic strain, which not only meets the requirements of high safety production, but also has relatively mature fermentation technology, simple fermentation culture requirements, and can also meet the industrial application conditions of simple fermentation operation and low cost. However, the current Bacillus clausii is mostly used in the industrial production of protease, and no relevant reports have been found on its use in the industrial production of ectoine

Method used

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  • Bacillus clausii and method for producing tetrahydropyrimidine by using same
  • Bacillus clausii and method for producing tetrahydropyrimidine by using same
  • Bacillus clausii and method for producing tetrahydropyrimidine by using same

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Embodiment 1

[0039] The acquisition of embodiment 1 bacillus clausii ET01

[0040] Take 1g of the mud sample from the Weihai offshore salt field and dilute it to 100mL with sterile water, apply 200μL of the diluted solution on 20g / L LB solid medium, place it in a constant temperature incubator at 37°C for 24 hours, and pick a single colony for several rounds Streak culture, finally isolated to obtain purified strains.

[0041] Bacterial morphological identification of the strain was carried out. After cultivating the bacterial strain at 38°C for 24 hours on the LB solid medium of 10g / L NaCl, milky white colonies were formed, with irregular edges and protrusions on the surface; observed under a microscope at 100 times, the thalline was in the shape of a long rod (such as figure 1 Shown), positive by Gram staining, for Gram-positive bacteria. According to the bacterial morphology and 16S rDNA sequencing comparison, the strain was identified and named as Bacillus clausii ET01 (Bacillus clau...

Embodiment 2

[0042] The culture condition and product detection of embodiment 2 bacillus clausii

[0043] The Bacillus clausii ET01 preserved in the glycerol tube was inoculated in 60 mL of 10 g / L NaCl LB medium at an amount of 2 v / v %, and cultured and activated overnight at 37° C. on a shaker at 200 rpm. Use a spectrophotometer to measure the absorbance (OD) of the fermentation broth at a wavelength of 600 nm. 600 ) is 2.3.

[0044] The activated Bacillus clausii ET01 was respectively inoculated in 60 mL of LB culture medium containing 0 g / L, 30 g / L, 50 g / L, 70 g / L and 100 g / L NaCl with the amount of 1 v / v%. Incubate on bed at 37°C, 200rpm for 24h.

[0045] In order to detect the growth situation of Bacillus clausii ETO1 of the present invention under each salt concentration, use spectrophotometer to measure the OD of fermented liquid after cultivating 24h 600 . The fermentation products (tetrahydropyrimidine and hydroxypyrimidine) in the fermentation broth of Bacillus clausii cultiv...

Embodiment 3

[0058] The cultivation and product detection of embodiment 3 bacillus clausii TDB1-4

[0059] Bacillus clausii TDB1-4 (CCTCC NO: M2015430) and Bacillus clausii ETO1 of the present invention were used for fermentation in LB medium with NaCl concentrations of 0 g / L and 50 g / L, respectively. The bacterial strain is cultivated according to the method of Example 2 and the OD after fermentation and cultivation for 24 hours 600 and ectoine were detected, and the results are shown in Table 2.

[0060] Table 2 The OD of Bacillus clausii ET01 and TDB1-4 600 and ectoine test results

[0061]

[0062] As can be seen from Table 2, under the situation of adding without NaCl, Bacillus clausii ET01 of the present invention is similar to the growth situation of bacillus clausii TDB1-4; But in the LB substratum of 50g / L NaCl, the present invention The Bacillus clausii ET01 grew well and produced ectoine; whereas the Bacillus clausii TDB1-4 grew poorly and did not produce ectoine. This sh...

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Abstract

The invention relates to bacillus clausii ET01 capable of producing tetrahydropyrimidine. Bacillus clausii ET01 is preserved in China General Microbiological Culture Collection Center (CGMCC), and thepreservation number is CGMCC No. 16310. The invention also relates to a method for producing tetrahydropyrimidine by using the strain for fermentation. The bacillus clausii disclosed by the inventioncan generate tetrahydropyrimidine with relatively high yield under the stimulation of low salt concentration, can be used for producing tetrahydropyrimidine through industrial fermentation, and has agood application prospect.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular, the invention relates to a strain of Bacillus clausii producing ectoine, and the application and method of using the strain to produce ectoine. Background technique [0002] Ectoine is a compatible solute produced in cells by many salt-tolerant and halophilic microorganisms to maintain osmotic pressure balance. , proteins, cell membranes and nucleic acids provide protection. In addition, ectoine has a certain effect on neurological diseases such as Alzheimer's disease and Parkinson's disease, and the latest research has found that ectoine can improve skin regeneration and delay skin aging. Therefore, ectoine will have broad application prospects in industries such as fine chemical industry and biomedicine. [0003] At present, the production method of ectoine is mainly obtained by high-density fermentation of halophilic microorganisms (especially Halomonas). Generally speaking, thi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/12C12R1/07
CPCC12P17/12C12N1/205C12R2001/07C12N1/20
Inventor 谭剑邓莉川于淼王靖王小艳殷红田芳王灿许克家江俊杰李榕榕陈博
Owner BEIJING BIOINNO BIOTECHNOLOGY CO LTD
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