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Antibacterial Macrolactin A that bacillus polyfermenticus KJS-2 produced in

A technology of macrolide and bacillus, applied in the field of antibiotics, can solve the problems of low yield, difficulty in identification, hindering industrial application, etc., and achieve the effect of broad-spectrum antibacterial activity

Inactive Publication Date: 2010-01-27
INJE UNIV IND ACADEMIC COOP FOUND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These results are very meaningful, however the maximum amount of macrolide A purified from various strains was less than 1 mg / l, and the maximum amount of malonyl-macrolide A (MMA) less than 1.2mg / l, which has hindered their industrial application
In addition, optical isomers of macrolide A have not been studied and their low yields make their identification difficult

Method used

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  • Antibacterial Macrolactin A that bacillus polyfermenticus KJS-2 produced in
  • Antibacterial Macrolactin A that bacillus polyfermenticus KJS-2 produced in
  • Antibacterial Macrolactin A that bacillus polyfermenticus KJS-2 produced in

Examples

Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1: the separation and identification of polyzyme bacillus KJS-2 and the generation of antibacterial substance

[0049] [Step 1: Isolation and Identification of Polyzyme Bacillus KJS-2]

[0050] During the antibacterial activity test of Polyzyme Bacillus isolated in 1933 by Dr. Terakado's research group in Japan, a strain was isolated that was morphologically different from other Bacillus strains. Microscopic observation showed that the strain had the characteristics of Bacillus and produced spores, and pedigree analysis based on the DNA sequence homology of 16s rRNA confirmed that it was a new strain belonging to the genus Bacillus. The present inventors confirmed that the base sequence of the 16s rRNA of the strain of the present invention is identical to that of the 16s rRNA of Bacillus PP19-H3 producing the previously known macrolide A (Korean Patent Application No. 10-2006096935: 2006.10.02) have 99% homology in their base sequences.

[0051] [Table 1] ...

Embodiment 2

[0063] Embodiment 2: purify antibacterial substance from the culture medium of polyzyme bacillus KJS-2

[0064] In order to purify the antibacterial substances produced by bacterial strain Bacillus polyzyme KJS-2, the seed culture of this bacterial strain was diluted in 3L of TSB medium (TSB agar: tryptone 17g, soytone 3g, glucose 2.5g, NaCl 5g, phosphoric acid dipotassium hydrogen 2.5g, pH6.8~7.2) to a final concentration of 4%, and cultivated for 2.5 days (30°C, 200rpm, 1vvm, pH6.8). The medium was extracted with ethyl acetate and analyzed by HPLC using the solvent of step 2 of Example 1 as Figure 4 Each peak was fractionated as shown in . Each fraction was tested for antibacterial activity against Escherichia coli, Bacillus subtilis 168 and vancomycin-resistant enterococci. The results showed that fractions 1, 4, 5 and 7 had antibacterial activity against Escherichia coli (see Figure 4 ), fractions 1, 2, 4, 5, 6, 7 and 9 have antibacterial activity against Bacillus sub...

Embodiment 3

[0065] Example 3: Structural analysis of fractions showing excellent antibacterial activity against VRE

[0066] In order to analyze the structure of the substance that inhibits the growth of Escherichia coli, Bacillus subtilis 168 and vancomycin-resistant enterococcus of the final purification, a large number of fractions were prepared under the same conditions as the preparative LC of step 2 of Example 1 (see Figure 6 ). Fractions were analyzed under the same conditions as the LC / Mass of Step 2 of Example 1, and Fraction 1 of Example 2 was purified with a purity of 97.72% (see Figure 7 ). 30 mg of material purified by preparative LC was dissolved in 700 μl of solvent DMSO-d6 and tested for first and second NMR ( 1 H-NMR, 13 C-NMR, 90-DEPT, 135-DEPT, H-H COZY, HMQC, HMBC). The results of the NMR analysis are shown in Table 2 below. Figure 8 , Figure 9 , Figure 10 , Figure 11 , Figure 12 , Figure 13 and Figure 14 middle. Based on the results of NMR analysi...

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Abstract

The present invention relates to uses of Macrolactin A produced by Bacillus polyfermenticus KJS-2 (KCCM 10769P), which is a new bacillus strain, as an antibiotic. Macrolactin A of the present invention, which is produced by Bacillus polyfermenticus KJS-2, shows a broad spectrum of antibiotic activity against a variety of microorganisms and fungi, and is proved to be very efficient for the inhibition of particularly vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus Aureus (MRSA) that are multidrug-resistant bacteria. The antibiotic Macrolactin A produced by Bacillus polyfermenticus KJS-2, can be used as an excellent antibiotic against vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus Aureus (MRSA), and thus the present invention is a very useful invention for medical industry.

Description

technical field [0001] The present invention relates to a kind of antibiotic produced by polyzyme Bacillus (Bacillus polyfermenticus) KJS-2 (KCCM 10769P)---macrolactin A (MacrolactinA) and application thereof; In particular, it relates to anti-harmful bacteria such as vancomycin-resistant Macrolide A and its application for antibacterial activity against Veterinary Enterococcus (VRE) and Methicillin-resistant Staphylococcus Aureu (MRSA). Background technique [0002] The increase of vancomycin-resistant enterococci (VRE) is unfortunate for human beings and brings many problems, such as very high cost of development of new antibiotics. It has been reported that the VRE infection rate in hospitals was only 0.3% in 1989, but this rate increased to 7.9% in 1993. Bacteremia mortality due to multidrug-resistant VREs is as high as approximately 70%, and there are concerns about the ability of VRE resistance genes to transform other Gram-positive cocci, which may increase the likel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/02
CPCC12R1/07C12P17/08C07D313/00A61P31/04A61P31/12C12N1/205C12R2001/07C12P17/02
Inventor 姜在璿金天圭金东熙金江民金东勋李进英崔光进车仁俊洪才宪洪勇根
Owner INJE UNIV IND ACADEMIC COOP FOUND
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