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Alkaline protease producing strain in sea cucumber intestinal tract and application thereof

A technology of protease and bacterial strains, which is applied in the direction of bacteria, biochemical equipment and methods, microorganisms, etc., can solve the problems of resource destruction, poor development of the nutritional value of sea cucumber leftovers, pollution of the environment, etc., and achieve the effect of efficient production

Inactive Publication Date: 2014-06-18
福建省参露生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the overfishing of sea cucumbers has damaged many commercial species resources to varying degrees.
Moreover, during the processing of sea cucumber products, leftovers such as sea cucumber sausages are directly thrown away as waste, which not only pollutes the environment, but also has not well developed the nutritional value of sea cucumber leftovers.

Method used

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  • Alkaline protease producing strain in sea cucumber intestinal tract and application thereof
  • Alkaline protease producing strain in sea cucumber intestinal tract and application thereof
  • Alkaline protease producing strain in sea cucumber intestinal tract and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: Screening of Thalassobacillus sp.QDHF bacterial strain:

[0023] 1. Materials and medium:

[0024] 1. Material sea cucumber

[0025] 2. Medium

[0026] Selection medium: 2216E medium and VNSS medium (see literature Christer O. Intestinal Colonization Potential of Turbot(Scophthalmus maximus) and Dab(Limanda limanda)-Associated Bacteria with Inhibitory Effects against Vibrio anguillarum[J].Applied and enviromental microbiology, 1992(58):551-556).

[0027] Casein medium: 1% casein, 0.1% yeast extract, 1.5% agar, pH 7.2, prepared with seawater, the above is the mass concentration, that is, the casein is 1g / 100mL.

[0028] Fermentation medium: peptone 0.5%, yeast extract 0.1%, ferric phosphate 0.01%, medium initial pH 8.0, prepared with seawater.

[0029] Solid medium: Weigh 3.3g LB nutrient agar medium and dissolve it in 100ml seawater.

[0030] 2. Screening of sea cucumber intestinal protease-producing bacteria:

[0031] The main reason for choosing th...

Embodiment 2

[0060] Embodiment 2: Optimal fermentation conditions of Thalassobacillus sp.QDHF bacterial strain

[0061] 1. Effect of different carbon sources on protease production by Thalassobacillus sp.QDHF strain

[0062] Different carbon sources, namely yeast extract, lactose, sucrose, starch, fructose and glucose with a mass concentration of 0.5 g / 100 mL were used in the liquid medium in Example 1. The culture condition is 25 DEG C, 180r / min shaking flask culture 3 days, fermented liquid is at 4 DEG C, centrifugal 15min under the condition of 6000r / min, removes bacterium, obtains supernatant, according to " three, protease activity measuring method in embodiment 1 "Measuring the protease activity of the supernatant. The results showed that the enzyme activity was the highest (701.8U / mL) when the yeast extract was used as the carbon source, and it was found that sugar significantly inhibited the growth and enzyme production of the strain (see figure 1 ).

[0063] 2. Effects of diffe...

Embodiment 3

[0078] Example 3: Preparation of sea cucumber polypeptides by fermenting sea cucumber scraps with Thalassobacillus sp.QDHF strain

[0079] Thalassobacillus sp.QDHF was inoculated in 2216E liquid medium (initial pH 8.0), and cultured at 30°C and 180r / min for 12 hours as a seed solution.

[0080] 1. Determination of peptide content

[0081] Protein quantification kit (BCA method) was used to determine the peptide content.

[0082] Prepare working solution A and working solution B in the kit at a ratio of 50:1 to prepare 200 μL of a mixed solution. Take 20 μL of the sample and add it to the above mixture and mix thoroughly. After bathing in water at 37° C. for 1 hour, measure the absorbance of the solution at 562 nm. The standard curve of peptide content and absorbance obtained with bovine serum albumin as a standard is: peptide content (mg / mL) = 0.53×A 562nm —0.0017.

[0083] 2. Effect of inoculum amount on preparation of polypeptide from fermented sea cucumber waste

[0...

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Abstract

The invention provides an alkaline protease producing strain (Thalassobacillus sp.) separated from a sea cucumber intestinal tract. The alkaline protease producing strain has been preserved in the general microbiology center for China Committee for Culture Collection of microorganisms of microbiology research institute of China science academy in Beijing City, Chaoyang District No. 1 Beichen West Road, No. 3 courtyard at October 18th, 2013, and the preservation number is CGMCC No.8366. The strain is bacillus, has the colony characteristics of being wet and faint yellow, and is a circle of which the center is convex and the edge is uniform. The invention discloses main components, mass concentration and a cultivation technology of an enzyme fermentation medium. The alkaline protease producing strain is applied to enzymolysis of sea cucumber processing scraps by a fermentation method, so as to prepare a bioactive polypeptide. Therefore, the alkaline protease producing strain has good popularization and application prospects.

Description

technical field [0001] The invention belongs to the technical field of functional microorganism screening, and in particular relates to a sea cucumber intestinal alkaline protease-producing strain and application thereof. Background technique [0002] Sea cucumber (Sea cucumber) is rich in a large number of bioactive components, such as mucopolysaccharides, saponins, β-carotene, sea urchin perinone, taurine, etc., and has extremely high nutritional and medicinal value. The increasing consumer demand for sea cucumber in the international market has caused the price of sea cucumber to rise all the way, which greatly stimulated the development of sea cucumber fishing industry. However, the overfishing of sea cucumbers has damaged many commercial species resources to varying degrees. And in the processing of sea cucumber products, leftovers such as sea cucumber sausages are directly thrown away as waste, which not only pollutes the environment, but also has not well developed t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P21/00C12R1/01
Inventor 朴美子陈勇王莹
Owner 福建省参露生物科技有限公司
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