Method for cultivating schizochytrium oil-producing grease employing schizochytrium and crude glycerine

A technology of Schizochytrium and crude glycerin, applied in the field of microorganisms

Inactive Publication Date: 2015-05-06
WENZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research in this area has just started, and there are few domestic reports

Method used

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  • Method for cultivating schizochytrium oil-producing grease employing schizochytrium and crude glycerine
  • Method for cultivating schizochytrium oil-producing grease employing schizochytrium and crude glycerine
  • Method for cultivating schizochytrium oil-producing grease employing schizochytrium and crude glycerine

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Experimental program
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Effect test

Embodiment 1

[0040] Embodiment one: the pretreatment of biodiesel by-product crude glycerin

[0041] Crude glycerin, a by-product of biodiesel, is obtained from biodiesel production enterprises, and is used to produce biodiesel from kitchen waste oil.

[0042] After mixing crude glycerol, a by-product of biodiesel, with water at a ratio of 1:4 (V:V), adjust the pH to 6.5, fatty acids form fatty acid salt (soap) precipitation, centrifuge at 4000 rpm for 10 minutes to remove soap to obtain crude glycerin, and then Sterilize with high pressure steam for 10 minutes (to be sterilized after the medium is prepared when it is used to prepare the medium in the future) to remove methanol.

[0043] The pretreated crude glycerin was analyzed for relevant components, and the results are shown in Table 1. Minerals were measured by atomic absorption spectrophotometry, glycerol content was measured by potassium periodate method (Li Sumin, Chinese Journal of Blood Transfusion, 1999, 12(3):179~179), fatty ...

Embodiment 2

[0046] Embodiment two: Mutation breeding of oil-producing Schizochytrium

[0047] The starting strain is Schizochytrium sp. WZU6961 (the source of genetic resources is disclosed in the patent "Method for High-density Fermentation of Schizochytrium sp. to Produce DHA (Application No.: 201210338013.0)"). Obtained isolated from decaying leaves of woods.

[0048] Get the preserved bacterial strain (2.0ml thawed bacterial liquid) and inoculate in 50ml liquid culture medium (medium formula (g·L -1 ): crude glycerin 30 (actual glycerol after conversion), yeast powder 10, NaCl 15, KCl 0.5, MgSO 4 ·7H 2 O2.0, K 2 SO 4 0.65, KH 2 PO 4 1.0, (NH 4 ) 2 SO 4 1.0, CaCl 2 2H 2 O 0.17, MnCl 2 4H 2 O0.003, ZnSO 4 ·7H 2 O 0.003, CoCl 2 ·6H 2 O 0.00004, Na 2 MoO 4 2H 2 O 0.00004, CuSO 4 ·5H 2 O 0.002, NiSO 4 ·6H 2 O 0.002, FeSO 4 ·7H 2(0.01, pH natural), cultivated at 30°C, 150-160rpm for 72h to activate the strain, then transferred to a new liquid medium at 5% volume ra...

Embodiment 3

[0051] Example 3: Optimization of culture conditions for oil production by culturing Schizochytrium with crude glycerin

[0052] Inoculate the preserved bacterial strain (2.0ml thawed bacterium liquid) in the 250ml Erlenmeyer flask that 50ml seed culture medium (formula is the same as Example 2) is housed, cultivate 72h to activate under 30 ℃, 150~160rpm; Then press 5% The volume ratio is transferred in the 500ml Erlenmeyer flask that 100ml seed culture medium is housed, cultivates 48h to obtain seed at 30 ℃, 160rpm; Then transfer to the 100ml fermentation medium that is housed by 5% volume ratio (basic formula is the same as seed cultivation Base) in the 500ml Erlenmeyer flask, at a certain temperature, 150 ~ 160rpm under cultivation 72h, measure biomass, thalline fat content, assay method is the same as embodiment two.

[0053] The effects of nitrogen source types, growth factors, pH, and temperature on the growth and oil accumulation of Schizochytrium were mainly discussed....

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Abstract

The invention discloses a method for cultivating schizochytrium oil-producing grease employing schizochytrium and crude glycerine, and belongs to the technical field of microorganisms. The schizochytrium (Schizochytrium sp.) 691Y01 is preserved at the China General Microbiological Culture Collection Center; the accession number of the preservation center is CGMCC NO.9043; and the bacterial strain is obtained by ultraviolet mutagenesis of schizochytrium (Schizochytrium sp.) WZU6961. Compared with the Schizochytrium sp. WZU6961, the palmitic acid content is improved by 50% while the DHA is reduced by 30%; rapid growth and accumulation of grease are achieved by biodiesel byproduct crude glycerine as a carbon source; and grease components are relatively suitable for production of the biodiesel.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a method for culturing the schizochytrium and crude glycerin to produce oil from the schizochytrid. Background technique [0002] With the increasing population and the rapid development of industry, the excessive consumption of energy, the pollution of the environment and the excessive consumption of resources have forced human beings to seek renewable and clean energy that can replace petroleum, so biodiesel (fatty acid) Methyl ester) has attracted people's attention because of its environmental protection, renewability, and good flammability. In order to prevent global warming and reduce greenhouse gas emissions, the European Union, Brazil, and Canada have introduced policies to encourage the production of renewable, carbon-neutral biodiesel. Therefore, biodiesel has developed rapidly in recent years, exceeding 13.03 billion liters in 2010. Although my country's...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/64C12R1/645
CPCC12P7/649C12N1/145C12R2001/645Y02E50/10
Inventor 姚瑶周茂洪万晓云吴论文杨美慧
Owner WENZHOU UNIVERSITY
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