Method for separating and screening high-temperature-resistant polyhydroxybutyrate generating bacteria
A polyhydroxybutyrate, separation and screening technology, applied in the biological field, can solve the problems of high fermentation substrate prices, extraction and purification costs, and high production costs, reduce equipment investment and fermentation substrate costs, and reduce production processes and equipment. Requirement, the effect of increasing biomass
Inactive Publication Date: 2016-01-27
陆祖军
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[0003] At present, the production methods of PHB include microbial synthesis and transgenic plant synthesis; transgenic plant synthesis of PHB has not been successful in reducing production costs; the production cost of using microbial synthesis of PHB is relatively high, mainly due to high fermentation substrate prices and extraction and purification costs
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[0030] The screening method of high-temperature-resistant polyhydroxybutyrate-producing bacteria of the present invention comprises the following steps:
[0031] (1) Collect soil samples:
[0032] Sampling areas: ① North (Tianjin, Shandong), ② South (Guangxi, Guangdong).
[0033] Soil types: orchards, gardens, vegetable gardens, fields, hills, riversides, residential and industrial areas.
[0034] Plot setting and sampling: 100m in various soil types 2 The square is the plot, draw a circle with the intersection point of the diagonal line as the center and half the length of the diagonal line as the radius, and the intersection point of the circle and the diagonal line is the sampling point. The soil at 5cm below the surface of each point was collected as samples, each sample was 1 kg, and each sample plot had 4 samples, a total of 32 samples;
[0035] (2) Sample pretreatment:
[0036] Take 10g samples respectively and put them into sterilized glass bottles filled with 90mL...
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The invention discloses a method for separating and screening high-temperature-resistant polyhydroxybutyrate generating bacteria. The method for separating and screening the high-temperature-resistant polyhydroxybutyrate generating bacteria includes the steps of sample acquisition, pre-treatment, amplification culture, stage-by-stage temperature rise screening, high-temperature-resistant flora selection, high-temperature-resistant strain selection and the like. The method has the advantages that the bacteria can resist high temperatures of 60 DEG C, so that the oxygen concentration can be quite conveniently controlled by means of controlling the temperatures, and requirements on production technologies and equipment can be lowered; PHB (polyhydroxybutyrate) can be grown and synthesized in simple fermentation equipment by taking cane sugar and cassava industrial wastewater with high organic matter contents as a carbon source, so that the equipment input and fermentation substrate costs can be reduced; and other non-high-temperature-resistant bacteria can be killed by means of increasing the temperatures, so that the uniqueness of the target bacteria can be guaranteed, and the biomass of the bacteria and the purity of the extracted PHB can be increased.
Description
technical field [0001] The invention relates to biotechnology, in particular to a method for separating and screening high-temperature-resistant polyhydroxybutyrate-producing bacteria (PHB). Background technique [0002] Polyhydroxybutyrate (polyhydroxybutyrate, PHB) is a kind of intracellular polyester, which is biodegradable and is a new type of polymer material for biodegradable plastics. [0003] At present, PHB production methods include microbial synthesis and transgenic plant synthesis; transgenic plant synthesis of PHB has not been successful in reducing production costs; the production cost of using microbial synthesis of PHB is relatively high, mainly due to high fermentation substrate prices and extraction and purification costs. [0004] The industry mainly adopts two-step method to cultivate microorganisms (aerobic / nitrogen-rich and oxygen-poor / nitrogen-poor) to obtain PHB. The method of controlling oxygen content is simple oxygen concentration control (mainly i...
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Inventor 陆祖军
Owner 陆祖军