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Bacillus amyloliquefaciens and application thereof in preparation of 1-deoxynojirimycin

A technology of amylolytic spores and bacillus, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of long fermentation cycle, large power consumption, long fermentation time, etc., and achieve industrial production and high production capacity High, short fermentation cycle effect

Active Publication Date: 2021-11-12
ZHEJIANG HUIDA BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, Yohji Ezure et al. (1985) mutated Streptomyces lavender to obtain the highest yield of DNJ, which can reach 4-5g / L. However, this strain is a filamentous fungus, and the fermentation process consumes a lot of power and the fermentation cycle is long.
CN105296565A discloses the use of Bacillus subtilis solid-state fermentation to obtain DNJ with lower purity
In summary, the microorganisms that can produce 1-deoxynojirimycin reported in the prior art generally have the disadvantages of low 1-deoxynojirimycin yield and long fermentation time.

Method used

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  • Bacillus amyloliquefaciens and application thereof in preparation of 1-deoxynojirimycin
  • Bacillus amyloliquefaciens and application thereof in preparation of 1-deoxynojirimycin
  • Bacillus amyloliquefaciens and application thereof in preparation of 1-deoxynojirimycin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 departure bacterial strain source

[0027] The original DNJ-producing strain was isolated from soil samples of mulberry roots in Xiniuqiao Village, Tongxiang City, Zhejiang Province.

[0028] Collect 20g of mulberry rhizosphere 3-10cm deep soil samples from different locations in Xiniuqiao Village, Tongxiang City, Zhejiang Province, put them into sterile sample bags, number them, and record the sampling locations and dates. All soil samples were passed through a 60-mesh sieve, and 1 g of each soil sample was added to sterile saline with glass beads. After fully shaking, put it in a water bath at 80°C for 40 minutes, and after standing for 10 minutes, take the supernatant for gradient dilution. , take 0.1mL of 10-2-10-6 gradient dilution solution to inoculate and spread on nutrient agar medium respectively, incubate upside down at 37°C for 2-5 days, and observe every day. Select the white bacterial colony, and after two rounds of streaking and passage purif...

Embodiment 2D

[0032] Embodiment 2 DNJ original production strain (HDCC00031) morphological examination and physiological and biochemical tests

[0033]Streak inoculation of strain (HDCC00031) on nutrient agar plate, culture at 37°C, record the characteristics of colony shape and color, and observe with optical microscope after Gram staining. Physiological and biochemical characteristics, glucose fermentation test, starch hydrolysis, V-P determination, indole test, gelatin hydrolysis, catalase test, nitrate reduction test, gas production test, anaerobic agar test were carried out on the target strain respectively.

[0034] Morphological characteristics: The colonies are irregular round or oval, white, with rough surface and bulges. Colony morphology such as image 3 As shown, the microscopic examination photos are as follows Figure 4 shown.

[0035] Physiological and biochemical characteristics: see Table 1 to Table 4 for details.

[0036] Table 1 The carbon source and nitrogen source u...

Embodiment 3D

[0045] Example 3 DNJ original production strain (HDCC00031) 16S rDNA identification

[0046] Take the slant of the original strain (HDCC00031), collect the fresh bacterial lawn, use Shanghai Sangon’s SK8255 kit to extract the DNA genome, and use universal primers (27F and 1492R) to amplify the 16S rRNA gene. After the PCR product is detected and purified, it is directly processed Sequencing was performed by the Institute of Bioengineering, Zhejiang University of Technology. The 16S rDNA sequence (SEQ ID NO: 1) measured by the bacterial strain (HDCC00031) was checked and compared with the sequences of related species and genus in the GenBank database by BLAST homologous sequence, as shown in Table 5 (only homologous It was found that the strain was very close to the taxonomic parameters of Bacillus amyloliquefaciens strain. Therefore, the strain HDCC00031 was identified as a strain of Bacillus amyloliquefaciens strain.

[0047] Homology of Table 5 strain (HDCC00031) and typica...

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Abstract

The invention discloses bacillus amyloliquefaciens HDCC00252, the preservation number of the bacillus amyloliquefaciens HDCC00252 is CGMCC No.22781. Meanwhile, the invention discloses a method for producing the 1-deoxynojirimycin through fermentation culture of the bacillus amyloliquefaciens HDCC00252, the titer can reach 5.31 g / L or above, the fermentation period is short, and industrial production is facilitated.

Description

technical field [0001] The invention belongs to the technical field of industrial biology, and in particular relates to a strain of Bacillus amyloliquefaciens capable of producing 1-deoxynojirimycin, and a method and application for fermenting and producing 1-deoxynojirimycin using the strain. Background technique [0002] 1-Deoxynojirimycin (1-Deoxynojirimycin, DNJ) is a piperidine alkaloid, the chemical name is 3,4,5-trihydroxy-2-hydroxymethyltetrahydropyridine, which is present in plants, microorganisms And the natural sugar analogues in silkworm body, the highest content in mulberry tree in nature, is a powerful sugar metabolism enzyme inhibitor (such as α-glucosidase, hexokinase, glucuronidase and glycogen phosphatase, etc.), It can significantly delay the degradation process of polysaccharides, reduce the peak of postprandial blood sugar, and stabilize fasting blood sugar. Moreover, it also has the effects of losing weight, increasing the sensitivity of islets, anti-v...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/12C12R1/07
CPCC12N1/20C12P17/12
Inventor 彭湘屏郑玲辉朱进伟孙琼高祥石磊张敏陈世敏汪超
Owner ZHEJIANG HUIDA BIOTECHNOLOGY CO LTD
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