Lodderomyces elongisporus, as well as Se-accumulating culture method and application thereof
A technology of longevity yeast and a culture method, which is applied in the field of microorganisms and achieves the effects of high selenium-enriching efficiency, good application prospects and good selenium-enriching performance.
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Embodiment 1
[0028] real materials and methods
[0029] 1.1 Test material
[0030] 1.1.1 Strains
[0031] In the present invention, the inventor uses a selective medium to screen the Loudtia longidosporum isolated from the intestinal tract of freshwater fish, and analyzes the ITS sequence and hsp78 gene. The new Lodderomyces elongisporus is classified as Lodderomyces elongisporus, numbered SCLE01, and has been deposited in the China Center for Type Culture Collection (CCTCC) on May 15, 2015, with the preservation number CCTCC NO: M 2015301.
[0032] The morphological characteristics of the strain of L. elongisporus SCLE01 (L.elongisporus SCLE01, preservation number is CCTCC NO: M 2015301) described in the present invention are: the colony is milky white, slightly rough, and after overnight culture on solid medium, the colony is 0.5 small -1mm, the surface is relatively smooth, the edges are neat, the colony is raised, and the density is high; after static culture in the liquid medium, th...
Embodiment 2
[0056] This example evaluates its selenium-enriching performance by studying the selenium-tolerant ability and selenium-enriched fermentation conditions of freshwater fish-derived Loudtia longans, and obtains the concentration of selenium in the medium, the way of adding selenium, the inoculation amount and the culture time through a single factor test The level range of 4 factors provides a basis for evaluating the feasibility of freshwater fish-sourced Loudia longans as a selenium-enriched yeast strain.
[0057] 2.1 Determination method of biomass and selenium content
[0058] 2.1.1 Biomass measurement
[0059] After the shaking culture was completed, the culture medium was taken, centrifuged at 6000rpm for 5min, and the bacterial cell precipitate was washed 3 times with distilled water, and the bacterial cell was collected, dried at 90°C to constant weight, and then weighed.
[0060] 2.1.2 Determination of selenium content
Embodiment 3
[0083] 3.1 Test material
[0084] The yeast used in this test is the freshwater fish intestinal strain that has been isolated and identified: Saccharomyces longus sporogenes (SCLE01, preservation number CCTCC NO.M 2015301). Japanese eel juveniles and complete feed (as basic feed) were purchased from Dongguan Yinhua Eel Breeding Co., Ltd. Lysozyme (LZM) was purchased from Nanjing Jiancheng Bioengineering Institute.
[0085] 3.2 Test method
[0086] 3.2.1 Preparation of selenium-enriched yeast
[0087]Prepare selenium-enriched yeast according to the previous test results, and the specific conditions are as follows: 10% inoculum size, 20 μg / mL selenium concentration (final concentration in the medium), added in 3 times (6h, 9h, 12h) in the logarithmic phase , the amount added in turn is 25%, 25%, 50%, 30°C, 190rpm shaking culture for 36h, to obtain selenium-enriched Loudia longans. Then the biomass of selenium-enriched yeast and the selenium content of bacteria were measured,...
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