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Strain SH-1 producing alginate lyase and application thereof
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A technology of alginate lyase and bacterial strain, applied in the direction of lyase, application, bacteria, etc., can solve the problems of low enzyme activity, limiting the development of seaweed extract, single degradation site, etc.
Active Publication Date: 2019-09-10
SOUTH CHINA AGRI UNIV
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Problems solved by technology
However, the microbial-sourced alginate lyases still have defects such as low enzyme activity and single degradation site, which limit the development of enzymatic hydrolysis to prepare seaweed extracts.
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Embodiment 1
[0088] Example 1 Isolation, screening and identification of Microbulbifer sp.SH-1
[0091] (2) Primary screening medium: sodium alginate 10g / L, NaCl 15g / L, (NH4) 2 SO 4 5g / L, K 2 HPO 4 1g / L, MgSO 4 ·7H 2 O 1g / L, FeSO 4 ·7H 2 O 0.01g / L, pH 7.5. 1.8% (w / v) agar was added to the solid medium.
[0092] (3) The composition of the re-screening medium is the same as that of the primary screening medium, except that agar is not added.
[0093] 2. Screening of strains
[0094] (1) Enrichment: Weigh 5g of sandy soil collected near the seaweed breeding area in Fujian and mix with 100mL sterilized water evenly. Draw 1 mL of the above-mentioned sand and sterilized water and mix the supernatant to inoculate in 50 mL / 250 mL enrichment medium, and shake the fla...
Embodiment 2
[0106] Example 2 Culture Condition Research of Microbulbifer sp.SH-1
[0107] (1) Inoculate the Microbulbifer sp.SH-1 preserved in glycerol on the seed medium (sodium alginate 10g / L, peptone 5g / L, yeast extract 5g / L, sodium chloride 10g / L, pH 7.0), in Shake culture at 28°C and 180rpm for 12h to obtain the logarithmic growth phase (OD 600 =0.6~0.8) seed solution.
[0108] (2) Inoculate the seed solution obtained in step (1) into a liquid fermentation medium (sodium alginate 10g / L, ammoniumsulfate 5g / L, magnesiumsulfate 0.2g / L, potassium dihydrogen phosphate 1g / L, sulfurous acid Iron 0.02g / L, sodium chloride 5g / L), respectively study the pH (4.5,5.5,6,6.5,7,7.5,8), culture temperature (25,28,32,35,40 ℃), liquid volume (15, 30, 50, 60, 75, 90mL, container volume is 250mL), inoculum volume (1, 2, 3, 4, 5% (v / v), shaker speed (120, 150, 180, 210, 240rpm) and other culture conditions on the enzyme production of strain SH-1.
[0109] The result is as Figures 12 to 16 It was s...
Embodiment 3
[0110] Example 3 Preparation of alginate lyase by Microbulbifer sp.SH-1
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Abstract
The invention provides a strain SH-1 producing alginate lyase and application thereof. The strain SH-1 producing alginate lyase is named Microbulbifer sp. SH-1 and is preserved in China General Microbiological Culture Collection Center in Beijing on December 7, 2018, and the preservation number is CGMCC No. 16906. The strain can be used for producing alginate lyase by fermentation and can also bedirectly used for degrading brown algae. The invention also provides the alginate lyase with exonuclease activity; the alginate lyase is obtained by fermenting the strain SH-1, the enzymatic activityof the alginate lyase can reach 757.90 U / mL, the stability is good, and the activity is high. The invention also provides a preparation method of an algae extract; active ingredients in algae are retained, meanwhile, sodium alginate is efficiently degraded into active oligosaccharides, secondary metabolites of part of microorganisms are increased, the yield of the algae extract can reach 78-92%, and the preparation method has good industrial prospects.
Description
technical field [0001] The invention belongs to the technical field of microbial strains and their applications, and relates to a strain SH-1 producing alginate lyase and its application, in particular to a strain Microbulbifer sp.SH-1 producing alginate lyase and its application. Background technique [0002] Chemical fertilizers play an important role in maintaining soil fertility and improving crop yield and quality. However, long-term and large-scale application of chemical fertilizers leads to many problems such as soil degradation, water eutrophication, and air pollution, which seriously restricts the sustainable development of modern agriculture. As a natural biostimulant, seaweed extract is not only rich in various active ingredients such as alginic acid, seaweed polysaccharides, brown algae polyphenols, mannitol, amino acids, natural plant hormones and essential mineral nutrients for plants, but also has biodegradable, non-toxic Toxic, non-polluting and environmenta...
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