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A recombinant streptomyces lydicus producing natamycin, a construction method and applications thereof

A technology of Streptomyces liidi and natamycin, which is applied to the field of recombinant Streptomyces liidi that produces natamycin and its construction, can solve the problems of polluting the ecological environment, increasing the drug resistance of pathogenic bacteria, and being ineffective.

Active Publication Date: 2013-04-24
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

General chemical pesticides are difficult to work, and frequent use increases the resistance of pathogenic bacteria and pollutes the ecological environment. Therefore, it is urgent to develop new, environmentally friendly and efficient microbial pesticides, which has become a hot spot in the field of disease biocontrol in recent years.

Method used

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  • A recombinant streptomyces lydicus producing natamycin, a construction method and applications thereof
  • A recombinant streptomyces lydicus producing natamycin, a construction method and applications thereof
  • A recombinant streptomyces lydicus producing natamycin, a construction method and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0056] Embodiment 1, construct the recombinant Streptomyces lydidae of high-yield natamycin

[0057] 1. Construction of cellulase gene expression vector pIB139-slnM

[0058] The name of the expression vector of the natamycin synthesis positive regulation gene constructed in this example is pIB139-slnM, and the vector contains the natamycin synthesis positive regulation gene slnM. The sequence of the natamycin synthesis positive regulatory gene slnM is shown in SEQ ID No.2. SEQ ID No.2 consists of 1019 nucleotides, the 1st-13th position is the NdeI recognition site and the protective base, the 14th-432nd position is the erythromycin resistance gene promoter, and the 433-1011th position is Na In the coding sequence of the gene positively regulated by tamycin synthesis, 1012-1019 are EcoRI recognition sites and protective bases.

[0059] The promoter in pIB139-slnM that initiates the transcription of the positively regulated gene for natamycin synthesis is the erythromycin resi...

Embodiment 2

[0118] Example 2. The antibacterial activity of Streptomyces lydicus AM02CGMCC No.6815 is higher than that of Streptomyces lydicus A02CGMCC No.1654

[0119] In this embodiment, the culture medium used is as follows:

[0120] Gao's No. 1 slant medium: K 2 HPO 4 0.5g, NaCl0.5g, KNO 3 1.0g, FeSO 4 ·7H 2 O0.01g, MgSO 4 ·7H 2 O0.5g, soluble starch 20g, agar 20g, water 1000ml; pH7.2-7.4.

[0121] Seed medium: 15g soybean grains (boil with distilled water for 0.5-1h, take the filtrate), 5g peptone, 2.5g ammonium sulfate, 20g glucose, 10g starch, 0.25g magnesium sulfate, 0.2g potassium dihydrogen phosphate, 5g sodium chloride, Make an aqueous solution, adjust the pH to 7-8, add 1g of calcium carbonate, and add water to make it 1000ml.

[0122] Fermentation medium: 15g soybean grains (boil with distilled water for 0.5-1h, take the filtrate), 5g peptone, 2.5g ammonium sulfate, 10g sucrose, 10g starch, 0.25g magnesium sulfate, 0.2g potassium dihydrogen phosphate, 5g sodium chlori...

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Abstract

The present invention discloses a recombinant streptomyces lydicus producing natamycin, a construction method and applications thereof. The construction method for the recombinant streptomyces lydicus producing natamycin includes a step of inducing a positive regulatory gene for the synthesis of natamycin into treptomyces lydicus as a host bacterium and screening to obtain a recombinant streptomyces lydicus with a natamycin yield higher than the host bacterium streptomyces lydicus. The positive regulatory gene for the synthesis of natamycin encodes proteins related in a) or b): a) a protein with the amino acid sequence shown in SEQ IDNo. 1; or b) derived proteins of a) which are related with natamycin synthesis and obtained via substitution and / or deletion and / or addition of one or several amino acid residues in the SEQ ID No. 1. The recombinant streptomyces lydicus of the invention has significantly improved natamycin producting and antibacterial capabilities compared to the wild-type streptomyces lydicus, and can be applied to food, bio-energy and feed additives and so on.

Description

technical field [0001] The invention relates to a genetically engineered bacterium and its construction method and application, in particular to a natamycin-producing recombinant Streptomyces lydidis and its construction method and application. Background technique [0002] With the adjustment of crop planting structure and changes in cultivation methods, the occurrence and damage of many crop fungal diseases are becoming more and more serious, such as cabbage wilt (Fusarium oxysporum f.sp.conglutinans), wheat sheath blight (R.cerealis), Pathogens such as peach brown rot (M.fructicola), apple ring rot (B.berengeriana), and lily root rot (F.oxysporum) are also very harmful (Geng Lihua et al., 2009). General chemical pesticides are ineffective, and frequent use increases the resistance of pathogenic bacteria and pollutes the ecological environment. Therefore, the urgent need to develop new, environmentally friendly and efficient microbial pesticides has become a hot spot in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/76C12N1/21C12P19/62A01N43/90A01P3/00C12R1/465
Inventor 吴慧玲刘伟成燕继晔董丹刘霆卢彩鸽张殿朋张涛涛田兆丰卢向阳
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES