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Damu Oceanobacillus and application thereof

A marine bacillus and spore-producing technology, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2014-08-06
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Marine Bacillus ( Oceanobacillus ) was proposed by Jie Lu and Yuichi Nogi et al. in 2002. The initial strain was isolated from the deep sea. Up to now, there have been 16 effective species and 2 effective subspecies. The concentration has a high tolerance, and there are few related reports on its application

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  • Damu Oceanobacillus and application thereof
  • Damu Oceanobacillus and application thereof
  • Damu Oceanobacillus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment one: Damum marine bacillus Oceanobacillus damuensis screening and separation

[0018] Sample collection and strain screening and isolation

[0019] The samples were collected in Shache County, Xinjiang Uygur Autonomous Region. The samples come from multiple areas, distributed between 76°1'57" to 77°46'30" east longitude and 37°27'30" to 39°15" north latitude. The soil 5-10cm from the surface was collected in a sterile glass bottle, air-dried, ground, sieved, and stored at 4°C for subsequent separation. The screening medium was phenol-enriched medium (PT), humic acid-enriched medium (HVA), sodium molybdate-enriched medium (HNA) and high-salt-enriched medium (YCSS), and the purification medium was pancreatic Peptone Soy Broth (TSB+10% NaCl), strains were screened by inoculating different enrichment media. Add 1 g of the soil sample into a shaker flask filled with 25 ml of liquid medium, and culture it with shaking at 28°C for 7 days. Then use a pipette gu...

Embodiment 2

[0028] Embodiment two: Damum marine bacillus Oceanobacillus damuensis multiphase taxonomic identification of

[0029] the strain Oceanobacillus damuensis PT-20 T The morphological observation, physiological and biochemical and cultural characteristics and chemical classification of the species were carried out in accordance with "Actinomycetes Systematics—Methods, Principles and Practice". Among them, the utilization of carbon source adopts the Biolog automatic bacterial identification system, and the high-performance liquid phase method is used to measure the quinone components and intracellular GC content of the bacteria. The determination of fatty acid components is analyzed by the Sherolock automatic bacterial identification system of American MIDI company. Using API ZYM reagent strips for identification, the reference strain is Oceanobacillus profundus CL-MP28 T (DSM 18246 T ), Oceanobacillus siheynsis , Ornithinibacillus bavariensis WSBC 24001 T , Carbon ...

Embodiment 3

[0031] Example 3: 16S rDNA sequencing and evolutionary tree construction

[0032] The total bacterial DNA was extracted by SDS method, the 16S rRNA gene sequence of the strain was amplified by PCR method, and the DNA sequencing was completed by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Go to the GenBank (http: / / www.ncbi.nlm.nih.gov / ) database to submit the partial sequence of the 16S rRNA gene measured by each strain for registration to obtain the sequence number. Then perform BLAST comparison on NCBI (National Center for Biotechnology Information) to find the strain with the closest relationship. The sequences of the corresponding strains with the closest relatives were compared by BioEdit, and then the Neighbor-Joinin (N-J) method was used to construct the phylogenetic tree on the software MEGA 5.0, and the bootstrap of the phylogenetic tree was 1000. The corresponding primers, reaction system and conditions for PCR are as follows: Primer PA 8 : 5'-CCGTCG...

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Abstract

The invention discloses (i) Oceanobacillus ( / i). (See accompanying drawing). Operations such as separation, screening and physiological and biochemical identification for saline-alkali soil in Yarkant County, Xinjiang prove that the strain is aerobic and Gram-positive, has motility, takes the shape of a short rod with a size of 0.2-0.4*1.0-1.2 mu m, spores grow at the ends, sporangium is slightly swollen, colony is of cream color, and is circular and regular, and the edge is smooth. The NaCl tolerance range is 3%-20% (the optimal salt concentration is 10%-15%), the pH growth range is 6.5-11.0 (the optimal pH range is from 7.5 to 9.0), and the temperature growth range is from 15 DEG C to 40 DEG C (the optimal growth temperature is 30-37 DEG C). The strain shows positive to oxidase and catalase, negative to urease, and the Oceanobacillus has nitrate reduction capability, can be used for hydrolyzing starch, and cannot be used for hydrolyzing tween-80, gelatin or cellulose. The Oceanobacillus can be widely applied to the production of amylase and high-salinity fermentation of various genetic engineering products. The accompanying drawing is a picture of PT-20 under a scanning electron microscope with 15000 times.

Description

technical field [0001] The present invention relates to the field of microbial strains and their application, specifically, the present invention relates to the technical field of marine bacillus and its application. Background technique [0002] Moderate halophilic bacteria is not a taxonomic term, but a general term for a class of extremophiles whose best salt-tolerant concentration is between 3% and 15%. It is widely distributed in various high-salt environments, such as oceans, salt lakes, saline-alkali lands, salt ponds, and many fermented foods. Due to the unique living environment of this bacteria, they have the following characteristics. Fermentation and growth at a higher salt concentration avoids the risk of bacterial infection during the fermentation process, does not require complicated culture conditions, and is physiologically and biochemically diverse. It has great potential application value. . [0003] In the traditional food fermentation industry, the app...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
Inventor 田永强刘波龙秀锋
Owner SICHUAN UNIV