Exiguobacterium profundum and application thereof

A technology of microbacteria and seeds in the deep sea, applied in the direction of application, bacteria, microorganisms, etc., can solve the problems of incomplete enzymatic hydrolysis, easy blockage of pipelines, increased costs, etc., to promote fish growth, reduce feed coefficient, and increase energy consumption costs Effect

Active Publication Date: 2020-04-17
浙江亿丰海洋生物制品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(1) In the preparation process of enzymatic fish paste, only endogenous enzymes are used, and the enzymatic hydrolysis is not complete; the addition of commercial enzymes such as protease increases the cost; after the enzymatic hydrolysis reaction, a high temperature treatment of about 90 ° C is required to destroy the enzyme, which will destroy the anchovy Functional active substances; enzymatic hydrolysis products have poor fluidity and are easy to block pipelines
(2) In the preparation process of ord

Method used

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  • Exiguobacterium profundum and application thereof
  • Exiguobacterium profundum and application thereof
  • Exiguobacterium profundum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Screening and identification of deep-sea microbacteria P-47-3

[0034] The process of screening high-temperature bacteria producing protease is as follows: figure 1 shown. Firstly, the water samples obtained from the Okinawa hydrothermal area in Japan were isolated by the dilution coating method of MA2216E medium (Difco, product number: 212185), and a total of 42 strains were isolated. Through temperature experiments, 12 strains that can continue to grow at 50°C were initially screened out. Adopt protease screening culture medium again, point inoculation flat plate, observe after cultivating 3-5 days, it is positive to form transparent circle, screen to 6 strains of protease-producing bacterial strains, wherein the transparent circle of P-47-3 bacterial strain is the largest (such as figure 2 shown).

[0035] The protease screening medium was prepared according to the following proportions: (1) Weighed 4 g of commercially imported skim milk (Difco, produc...

Embodiment 2

[0046] Embodiment 2: the property of protease produced by deep-sea microbacterium P-47-3

[0047] The protease activity was measured by the Folin-phenol method, and the Folin-phenol reagent was purchased from Solebol, Cat. No. F8060.

[0048] Step 1: Make a Standard Curve

[0049] Take 14 test tubes and divide them into two groups, add 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1mL standard protein solution (250μg / mL) respectively, make up to 1mL with water, add 5mL reagent A, mix well, and store at 20-25℃ Let it stand for 10 minutes, then add 0.5mL reagent B, shake it up immediately, keep it warm at 20-25°C for 30 minutes, and then measure the color at 500nm. Measure the optical density value, get the mean value (table 1) of two groups of measurements, take the protein concentration as the abscissa, the optical density value as the ordinate, and draw the standard curve value ( Figure 6 ).

[0050] Table 1: Casein Standard Curve Data Sheet

[0051]

[0052] Step 2: Inoculate Exobacte...

Embodiment 3

[0055] Embodiment 3: Utilize deep-sea microbacterium P-47-3 fermentation to produce fermented fish paste

[0056] Step 1: Preparation of cell liquid culture of deep-sea microbacterium P-47-3: take the strains stored at -80°C and streak on LB solid medium, culture at 40°C, pick a single colony and inoculate it in a 25mL LB medium In a 100mL Erlenmeyer flask with base, the culture temperature was 40°C, and cultured on a shaker at a speed of 150r / min for 20-24h to the mid-logarithmic growth phase, and the cell liquid culture of deep-sea Exiguobacterium P-47-3 was obtained;

[0057] Step 2: Shake flask fermentation culture: the inoculum size is 2-5% (v / v), inoculated in a 500mL Erlenmeyer flask with 200mL LB medium, the culture temperature is 40°C, and the rotation speed is 150r / min on the shaker Under culture for 20-24h to obtain a fermentation broth.

[0058] Step 3: continue the shake flask fermentation culture: the inoculum size is 2-5% (v / v), inoculated in 1000mL LB liquid m...

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Abstract

The invention provides an exiguobacterium profundum. The preservation number of the exiguobacterium profundum is CGMCC No.18990. The exiguobacterium profundum P-47-3 strain provided by the invention is used for fermenting fish paste. The strain P-47-3 used in the method is separated from a deep sea hydrothermal solution extreme environment; spores are produced, and the growth temperature ranges from 10 DEG C to 65 DEG C; growth and propagation can be performed under the salt concentration of 0 to 14%; general high-temperature bacteria mainly come from land environments such as hot spring and the like, are poor in salt resistance, can not grow at high salt conditions, and has great limitation in marine biological product fermentation, while the strain P-47-3 has halophilicity, so that the strain P-47-3 is not only suitable for marine organism fermentation, but also can consume salt in a metabolic process, the salt concentration in a fermentation product is further reduced, and the quality of the fermentation product is improved.

Description

technical field [0001] The invention belongs to the technical field of microbial screening application, and in particular relates to a thermophilic deep-sea microbacterium and its application. Background technique [0002] Fish meal is currently the main protein source in aquafeeds because of its high protein content, various essential amino acids, and easy absorption. China is a big aquaculture country in the world. In 2018, the total aquaculture output exceeded 50 million tons, accounting for more than 78% of my country's total aquatic product output. China is the country with the largest consumption of fishmeal. China's aquaculture industry consumes about 2.5 million tons of fishmeal (partially used for poultry farming) every year, of which 1.2 million tons are imported fishmeal and 1.3 million tons are domestic fishmeal. It can be seen that China's aquaculture industry relies on fishmeal resources and is also restricted by foreign fishmeal imports. [0003] In the pro...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/12C12R1/01
CPCA23K10/12C12N1/20C12N1/205C12R2001/01
Inventor 张德超许静媛王灵华许福土
Owner 浙江亿丰海洋生物制品有限公司
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